1-B-P-001

一般演題（ポスター）

The effects of dopaminergic agents on the high potassium-evoked DOPA anddopamine release from PC12 cells〇五嶋 良郎、青木 令奈、岡田 貴子、増川 太輝
横浜市立大・医L-3,4-Dihydroxyphenylalanine (DOPA), a precursor of a neurotransmitter dopamine (DA), is synthetized bytyrosine hydroxylase in the cytoplasm of catecholaminergic neurons. We proposed that DOPA is aneurotransmitter. We previously reported that high K+ -evoked release of DOPA and DA from cultured PC12
cells, both of which were similarly decreased by deprivation of extracellular Ca2+. Using this system, we areattempting to elucidate the mechanism by which the DOPA release occurs. We found that bafilomycin inhibited
the K+-evoked-release of DA, but not DOPA, while brefeldin A suppressed the release of DOPA but not DA,
thereby suggesting the release of DOPA may occur through a secretion pathway distinct from that for DA in
PC12 cells (JPS95). To further characterize the release of DOPA, we here examined the effects of several agentson the evoked release of DOPA and DA from cultured PC12 cells. Proline, valine, phenylalanine, tyrosine andalpha-methyl- -tyrosine (10-5 M) showed no effect on DOPA and DA release. Among reagents tested, 3-iodotyrosine inhibited the release of DOPA without affecting that of DA. This finding indicates a higher sensitivityof the release of DOPA to 3-iodo-tyrosine than that of DA.

Copyright © The Japanese Pharmacological Society1-B-P-002

一般演題（ポスター）

α-tocotrienol binds to 67 kDa laminin receptor, resulting in activation of DGKα.

〇難波 朋花、上田 修司、福田 伊津子、白井 康仁
神戸大・院農・生命機能科学専攻 応用生命化学講座

Vitamin E is classified into two types: tocopherols (Toc) and tocotrienols. Although their structures are very
similar, tocotrienols are known to have about 50 times the antioxidant capacity of Toc. Furthermore, it has beenreported to have anticancer activity against various types of cancer. Recently, we have reported that α-Toc andepigallocatechin gallate (EGCg) activate diacylglycerol kinase α (DGKα), which is a lipid kinase to convert
DG to phosphatidic acid, by direct binding to 67kDa laminin receptor (67LR) at respective different bindingsites, contributing to improvement of nephropathy. The binding of αToc or EGCg to 67LR has been shown to
induce palmitoylation of 67LR. These results suggest that α-tocotrienol also binds to 67LR and induces its
palmitoylation, resulting in DGKα activation. Indeed, in this study, we showed that α-tocotrienol activated
DGKα by binding to 67LR at the same site with α-Toc, and found that α-tocotrienol induced palmitoylationof 67LR with DGKα translocation (activation). In addition, the DGKα translocation rates of tocotrienol
homologues were compared and found that d-tocotrienol had similar effect with a-tocotrienol. These results
suggest that 67LR and subsequence activation of DGKa are involved in some functions of tocotrienols including
anticancer effects.

Copyright © The Japanese Pharmacological Society1-B-P-003

一般演題（ポスター）

Analysis of the molecular mechanism of neurite outgrowth under PACAP in PC12
cells.

〇山下 道生1、竹ノ谷 文子1、柴藤 淳子2、Rakwal Randeep3、平林 敬浩2、千葉 義彦4、塩田 清二51

星薬科大・薬・運動科学、2湘南医療大・臨床医療研、3筑波大・体育系、4星薬科大・薬・分子生物、5湘南医療大・薬・解剖生理We investigated the molecular mechanism of neurite outgrowth with PACAP treatments using a rat adrenalderived pheochromocytoma cell line-PC12. This study was specifically investigated into the regulation of
PACAP-induced CRMP2 previously identified in a mouse brain ischemia model and which could be recovered
by PACAP treatment. We have previously revealed that PACAP-mediated neuroprotection involves not only
CRMP2 but also pathways related to GSK-3β and other signaling components. To clarify whether CRMP2 acts
directly on PACAP or through GSK-3β as part of the mechanism of PACAP-induced neurite outgrowth, weobserved neurite outgrowth in the presence of GSK-3β inhibitors and activators. Post PACAP treatment in
PC12 cells, immunostaining was used to confirm protrusion elongation, while RT-PCR, 2D Gel with Western
blotting, and inhibition experiments were performed to confirm the expression of the PACAP gene, its
receptors, and downstream signaling components. These results indicate that neurite protrusion elongation by
PACAP follows a GSK-3β-regulated pathway through the PAC1-R, where the AKT and cAMP/ERK pathways
are involved. These findings also provide a solid basis for future research to develop new therapies andtherapeutic agents to treat neural disorders and may contribute to neurogenesis.

Copyright © The Japanese Pharmacological Society1-B-P-004

一般演題（ポスター）

Significance of melatonin production in mast cells〇西 晴久1、ニヨンサバ フランソワ2,3
1

東京慈恵会医科大・医、2順天堂大・院医・アトピーセ、3順天堂大・国際教養[Background and objective]
Mast cells, the cells responsible for both immediate and delayed allergic reactions, are also one of the few
melatonin-producing cells in the body. Melatonin, which is primarily released from the pineal gland andregulates circadian rhythm, has both hypnotic and antioxidant effects. However, the physiological function ofmelatonin produced by mast cells is unknown. We set out to investigate this phenomenon.
[Methods]
LAD2, a human-derived mast cell line, was used in all experiments. Melatonin production by mast cells was
detected by ELISA using anti-melatonin antibodies. mRNA and protein expression of melatonin synthase
proteins were analyzed by real-time PCR and Western blotting, respectively. Allergic stimulation of the cells was
performed by sensitizing the cells with 1.0 μg/mL anti-NP IgE, followed by stimulation with 1.3 μg/mL NPBSA.
[Results]
Expression of arylalkylamine N-acetyltransferase (AANAT) and hydroxyindole-O-methyltransferase (HIOMT),
key enzymes in melatonin production, was upregulated more than 2-fold at both mRNA and protein levels inmast cells treated by allergic-stimuli. The expression of the two enzymes was also upregulated by A23187, acalcium ionophore; however, the upregulation level was less than 2-fold for each enzyme. Conversely, mRNAand protein expression of both enzymes was slightly reduced in cells pre-treated with 500 μM db-cAMP, which
is known to increase cAMP.
[Discussion]
These results indicate during mast cell degranulation, representing an increase in intracellular calcium
concentration, the expression of melatonin-producing enzymes is increased during resynthesis of the granules.
However, IgE-mediated degranulation by allergic stimulation results in more production of melatonin than it isrequired for replacement, therefore contributing to the repair of damaged tissues.

Copyright © The Japanese Pharmacological Society1-B-P-005

一般演題（ポスター）

Antioxidant potential of histidine-rich glycoprotein contributes to improvementof sepsis pathophysiology〇和氣 秀徳1、森 秀治2、西堀 正洋3、ハティポール オメル ファルク1、西中 崇1、高橋 英夫1
近畿大・医・薬理、2就実大・薬、3岡山大・院医歯薬・創薬研究推進室

1

Histidine-rich glycoprotein (HRG) has been reported to have regulatory effects on blood coagulation andfibrinolytic systems. In addition to these effects, we found that HRG is a factor that is specifically downregulated
in sepsis, and that HRG contributes to the amelioration of sepsis via its effects on maintaining neutrophil
morphology and function and protecting vascular endothelial cells. The involvement of ROS in the progressionof septicemia has been reported, and in previous report, we have shown that HRG suppresses excess ROS
production from neutrophils, but the direct effects of HRG on ROS and ROS-producing systems are not known.
Therefore, in this study, we investigated the activity of HRG against ROS. HRG showed no effect on superoxide
and oxygen peroxide among ROS, but on hydroxyl radicals, which are highly toxic in vivo, it was found that
HRG suppressed the production of hydroxyl radicals by efficiently chelating the divalent iron ions used in the
Fenton reaction during the production of hydroxyl radicals. With regard to peroxyl radicals involved in lipid
peroxidation, which is important for cell damage, HRG exhibited antioxidant capacity even though it was not
mediated by the Fenton reaction, suggesting that in this system, the antioxidant capacity was due toautoxidation.

Copyright © The Japanese Pharmacological Society1-B-P-006

一般演題（ポスター）

Compound X is a novel ligand candidate for L-DOPA receptor GPR143〇井上 美優横浜市立大・院医・分子薬理神経生物

We propose that L-3,4-dihydroxyphenylalanine(L-DOPA) by itself is a neurotransmitter. Recently, a G-protein
coupled receptor GPR143, a gene product of ocular albinism 1, was identified as a receptor for L-DOPA.
However, L-DOPA is an unstable compound, and GPR143 ligands other than L-DOPA have not been identified
yet. In this study, we found compound X, a novel ligand for GPR143, and examined its pharmacological actions.
Using zero-maze test, we found that Gpr143 gene-deficient (Gpr143-/y) mice exhibited more anxiety-likebehavior when compared to wild-type (WT) mice. Intraventricular administration of compound X caused
anxiety-like behavior in WT mice. The effect of compound X was not observed in Gpr143-/y mice. These results
indicate that compound X showed an antagonistic activity against GPR143. Compound X may be useful toclarify the role of L-DOPA/GPR143.

Copyright © The Japanese Pharmacological Society1-B-P-007

一般演題（ポスター）

Cystathionine γ-lyase self-inactivates by generating supersulfides〇荒木 笙馬、土屋 幸弘、渡邊 泰男
昭和薬科大・薬・薬理学

Cystathionine γ-lyase (CSE) is an enzyme responsible for the biosynthesis of cysteine fromcystathionine. It also has the β-lyase activity toward cystine to generate cysteine hydropersulfide (Cys-SSH).
Notably, the chemical reactivity of Cys-SSH is thought to be involved in the catalytic activity of proteins via
protein polysulfidation. In the present study, we investigated Cys-SSH could regulate CSE enzyme activity
through its polysulfidation.
CSE enzyme activity (levels of enzymatically synthesized CysSSH) was determined with afluorescent probe sulfane sulfur probe 4 (SSP4). Polysulfidated CSE was detected using modified biotin switch
assay.
incubation of CSE with Na2S4 induced the inhibition of the enzyme,
accompanied by its polysulfidation. Treatment with dithiothreitol reversed the polysulfidation and thesubsequent inhibition. Similarly,

incubation of CSE with CSE-enzymatically synthesized Cys-SSH

resulted in the inhibition of the enzyme. We generated Na2S4-insensitive CSE mutants in that its specific Cys
resides were mutated with valine. Furthermore, the mutant displayed a reduction in CSE polysulfidation by
Na2S4 relative to the wild-type enzyme. Interestingly, the enzyme activities of Na2S4-insensitive CSE mutant
were higher than that of wild-type. Thus, CysSSH is endogenously generated and auto-inhibits CSE enzyme
activity via polysulfidation of its specific cysteine residues.

Copyright © The Japanese Pharmacological Society1-B-P-008

一般演題（ポスター）

Identification and Characterization of Human Colorectal Cancer Cluster
Predominantly Expressing EP3 Prostanoid Receptor Subtype〇福島 圭穣、藤野 裕道
徳島大・院医歯薬

Colorectal cancer (CRC) is one of the common types of cancer in humans. Prostaglandin E2 (PGE2) is a wellknown mediator of colorectal cancer through stimulation of four E-type prostanoid (EP) receptor subtypes:
EP1, EP2, EP3, and EP4 receptors. All subtypes of EP receptors are involved in CRC promotion or malignancy.
However, the characteristics of CRC that highly expresses EP receptor subtypes have not been clarified. In thepresent study, we classified CRC from a cancer genomic database and identified CRC clusters which highly
express EP receptor subtypes. Most of these clusters predominantly expressed one subtype of EP receptor andshowed different gene expression patterns. Among them, we focused on the cluster highly expressing the EP3
receptor (CL-EP3). As the result of characterization of gene expression, CL-EP3 was characterized as: epithelial
mesenchymal transition (EMT)-induced progressed cancer with activation of transforming growth factor-β
pathway, activation of hypoxia-inducible factor-1α, and suppression of runt-related transcription factor 3. Since
we previously reported that EP3 receptor is involved in and induce colon cancer cell migration, EP3 receptorexpressing CRC may induce metastasis through these signaling pathways. Thus, the findings suggest theeffectiveness of cancer clustering by gene expression of the EP receptor subtype to elucidate the mechanism of
human CRC.

Copyright © The Japanese Pharmacological Society1-B-P-009

一般演題（ポスター）

The recognition mechanism of the analgesic drug mirogabalin by recombinanthuman α2δ1 protein〇香西 大輔1,2,3、沼本 修孝4、西川 幸希5,6、亀川 亜希子3,5、川崎 祥平7、廣明 洋子1,2、入江 克雅1,8,9、大嶋 篤典1,8,10、半
沢 宏之7、島田 神生11、北野 裕11、藤吉 好則3,5
1

名古屋大・細胞生理研セ、2バイオ産業情報化コンソーシアム、3東京医科歯科大・高等研院、4東京医科歯科大・難治疾患研、5CeSPIA、6東京農工大・生体分子解析学共研講、7第一三共RDノバーレ、8名古屋大・院創薬科学、9和歌山県立医科大・薬、10名古屋大・iGCORE、11第一三共

Mirogabalin is a novel gabapentinoid drug, following gabapentin and pregabalin, and given as a first-line agent
for peripheral neuropathic pain. Mirogabalin is a γ-aminobutyric acid analog that binds to the voltage-gatedcalcium channel subunit α2δ1. However, the mirogabalin recognition mechanism of α2δ1 remains elusive.
Here we analyzed a structure of recombinant human α2δ1 bound to mirogabalin at 3.88 Å resolution by cryoelectron microscopy. Our structure and mutagenesis binding assays confirmed that the evidence shown byothers for gabapentinoid binding to a conserved amino acid binding motif located in the extracellular dCache_1
domain distal pocket applies to mirogabalin. In addition to the conserved residues interacting with the carboxyl
and amino groups of mirogabalin, our mutagenesis binding assays newly identified residues W205 and Y217 in
the hydrophobic interaction region are critical for mirogabalin binding. The A215L mutation, which wasintroduced to increase the side chain length at the hydrophobic interaction site of mirogabalin based on our
structure, predictably suppressed the binding of mirogabalin while promoting the binding of another ligand, LLeu, with a smaller hydrophobic substituent than mirogabalin. Our study indicates the significance ofhydrophobic interactions in α2δ1 mirogabalin recognition.

Copyright © The Japanese Pharmacological Society1-B-P-010

一般演題（ポスター）

Effects of the therapeutic drugs for Cystic Fibrosis in Caucasians on CFTR
mutations found in Japanese patients.

〇君島 莉央、相馬 光流、清水 正浩、大川 詩織、松澤 由佳、金子 すずな、深田 侑希、中尾 佳菜子、相馬 義郎
国際医療福祉大・院薬・分子病態治療学分野

Cystic fibrosis (CF) is an autosomal recessive disease in which mutations in the CFTR gene cause various
symptoms through its channel malfunction. Among over 1900 gene mutations, the most common mutation is
ΔF508 which causes a trafficking defect of CFTR to plasma membrane (classified as ʼclass IIʻ ). CF is quite rarein Japanese and listed in ʼIntractable Diseasesʻ by MHLW, Japan.
The most frequent point-mutations in Japanese patients are H1085R and L441P, both are class II same as
ΔF508. Recently the FDA granted Vertex Inc. approval for "Trikafta", a combination of two expression
correctors, elexacaftor and tezacaftor, and one channel function potentiator, ivacaftor. In Japan, at present, theCF treatment is limited to some symptomatic ones, and no radical treatment has not been approved yet. Trikafta
could be effective against disease-associated mutations other than ΔF508.
In this presentation, we will discuss about the therapeutic effects of Tricafta against H1085R and L441P, basedon ourdata.

Copyright © The Japanese Pharmacological Society1-B-P-011

一般演題（ポスター）

Molecular dynamics simulation study on structural instability of CFTR mutants inJapanese CF patients and effects of CFTR correctors on them.

〇相馬 光流、君島 莉央、石塚 柊太、中尾 香菜子、相馬 義郎
国際医療福祉大・院薬・分子病態治療学研究室

Cystic fibrosis (CF) is a genetic disorder caused by abnormal function of CFTR. CFTR is expressed in thelumen lateral membrane of the transport epithelia of the whole body, such as the gastrointestinal and therespiratory epithelia, and plays a central role in the anion transport. CF is common in Caucasians but relatively
rare in Asians. Deletion of phenylalanine at position 508 (ΔF508) is the most common CF-associated mutationand causes defects in CFTR trafficking to the plasma membrane (class II). The CFTR mutation profiles inJapanese are far different from those in Caucasians. Recently, some CFTR correctors rescuing the ΔF508CFTR from the trafficking defect were developed for by Vertex. Inc. We have confirmed that the Vertex
correctors for the ΔF508 mutant could also rescue Japanese CF-associated mutants, H1085R and L441P.
In this work, we study the molecular fluctuation of CFTR using molecular dynamics (MD) simulations because
the molecular instability might lead to the protein degradation. We will discuss about the mechanisms of theJapanese CF mutations causing the trafficking defect and how the Vertex correctors rescue them.
(COI:No)

Copyright © The Japanese Pharmacological Society1-B-P-012

一般演題（ポスター）

Trans-inhibitory effect of dotinurad, a uricosuric agent on uric acid reabsorptive
transporter URAT1〇藤田 一輝、朱 秋楠、荒川 大、白坂 善之、玉井 郁巳
金沢大・医薬保健研究域薬学系・薬物動態学

URAT1 is responsible for renal reabsorption of uric acid (UA) and is a target of uricosuric agents. Newly
developed uricosuric agent dotinurad selectively inhibits URAT1. Interestingly, in vitro experimentally obtained
Ki of dotinurad was larger than in vivo Ki estimated by model analysis of clinical uricosuric effect (Pharmacol
Res Perspect. 7(6):e00533, 2019). In the present study, we precisely analysed inhibition mechanism of URAT1
by dotinurad.

oocytes that were injected with cRNA of URAT1 were used for analysis of URAT1

activity. When dotinurad was preincubated with oocytes, UA uptake were decreased more strongly than without
preincubation. Furthermore, when dotinurad was directly injected into oocytes and immediately measured UA
uptake without dotinurad in the uptake medium, UA uptake was decreased, while such pre-injection effect was
not observed by conventional URAT1 inhibitor benzbromarone. These results demonstrated that dotinurad
specifically affects URAT1 from intracellular side (-inhibition) in addition to-inhibition. Accordingly,

clinical effect of dotinurad is explained by inhibition of URAT1 from both tubular lumen side (and intracellular side (-inhibition)-inhibition) and such dual mechanism may explain apparent difference of URAT1

inhibition potency between in

and in.

Copyright © The Japanese Pharmacological Society1-B-P-013

一般演題（ポスター）

Physiological and Anatomical Properties of Utricular Hair Cells and Afferents
del/delin Gpr156
Mice Lacking a Mirror-Image Hair Cell Organization〇小野 和也1,2、タチーニ バジル3、イトック ルーズアン2、日比野 浩1
大阪大・医・薬理学講座 統合薬理、2シカゴ大学、3ジャクソン研究所

1

In otolith organs, hair bundles of hair cells have varying orientations that reverse along the line of polarity
reversal (LPR) located within or at the edge of the central striolar zone. Despite such unique anatomical feature,
the significance of simultaneous excitatory and inhibitory inputs upon translational stimuli remains to beunderstood. GPR156 is a G protein-coupled receptor (GPCR) belonging to metabotropic glutamate receptor
subfamily. Recently, Kindt et al. has shown that GPR156 uniformly expressed in the hair cells of five vestibularorgans becomes polarized and reverses hair bundle orientation under the control of Emx2, a transcription factor,
establishing mirror-image hair cell organization in the otolith organs. (Nat Commun 2021).

otolithorgans lose the LPR without any other clear anatomical defects. In this study, we examined whether the loss of
LPR inmice affects physiological and anatomical properties of hair cells and primary afferent

neurons.
We found that the transducer currents of type I and II hair cells in null and het animals had comparable
properties (operating range, sensitivity, adaptation time course and extent).
We also determined whether zone-specific firing pattern is affected by loss of LPR. In wildtype animals, afferent
neurons innervating the striola and extrastriola including LES tend to fire in transient and sustained fashion,respectively, in response to depolarizing current steps. Our results show that current steps applied to LES
calyces of both

andunaltered firing pattern by loss ofutricles elicited multiple spikes (sustained responses), suggesting.

Together, we conclude thatsome physiological properties are conserved in hair cells and afferent terminals of
the LES in utricles without bundle orientation reversal as a result of deletion of

Copyright © The Japanese Pharmacological Society.

1-B-P-014

一般演題（ポスター）

Different reactivity of TRPM8 and TRPA1 to menthol in dogs

〇山口 卓哉1、柳川 日向子1、松岡 彩那1、内田 邦敏2、山﨑 純1
1

日本大・生物資源科学部・獣医薬理学研究室、2静岡県立大・食品栄養科学部・生体機能学研究室

Although menthol generally provides a pleasant cooling sensation in humans, at high concentrations it can cause
discomfort and/or pain. This effect is attributable to two cation channels, TRPM8 and TRPA1, that exhibit
different dose dependency in humans. Given that the dose dependency of these receptors to menthol might
differ in other mammalian species, we examined the reactivity of canine TRPM8 and TRPA1 to menthol byusing recombinantly expressed channels. HEK293T cells were transfected with canine TRPM8 or TRPA1 andsubjected to calcium imaging. Canine TRPA1 was confirmed to be activated by the selective TRPA1 agonist allyl
isothiocyanate in a dose-dependent manner, which was inhibited by the TRPA1 antagonist HC-030031. Canine
TRPA1 was also activated by menthol in a dose-dependent manner, even at high concentrations (up to 3 mM),
in contrast with TRPA1 in rodents, which was inhibited by a high concentration of menthol. Canine TRPA1
showed a much higher value at EC50 for menthol response compared with canine TRPM8 (178.9 μM and 5.3
μM, respectively). The activation of TRPA1 by menthol was inhibited by HC-030031 but it was not inhibitedby the TRPM8 antagonist RQ-00203078. Our results suggest that menthol activates TRPM8 at low
concentrations and may induce a pleasant cooling sensation, whereas at high concentrations it activates TRPA1
and may cause discomfort in dogs as in humans. In addition, this mechanism is suggested to be conserved insome carnivora and primates.

Copyright © The Japanese Pharmacological Society1-B-P-015

一般演題（ポスター）

Functional and structural basis of hERG facilitation by its blockers〇古谷 和春、河野 諒太朗、喜多 紗斗美
徳島文理大・薬Certain human Ether-à-go-go-Related Gene (hERG) blockers can facilitate hERG activation to increase hERG
currents, which may reduce proarrhythmic potential. However, the molecular mechanism involved in thefacilitation effect of hERG blockers remains unclear. Here, we demonstrate that 1) nifekalant accesses thereceptor site within the pore of the open or inactivated channels at depolarized potentials, 2) upon return to theresting potentials, channels close and trap nifekalant inside, 3) trapped nifekalant biases the open-closed
equilibrium towards the open state, and 4) the kinetics of drug escape from the channel are faster than channel
closing rates at potentials where facilitation of hERG current is observed, thereby drug unbinding reveals
channels that have been biased towards the open state. Simulations with a Markov model of such nifekalanthERG interaction successfully reproduce key characteristics of hERG facilitation. We also present a potential
structural model for hERG channel facilitation through drug interactions with the hydrophobic pocket of thehERG pore domain. This pattern of interaction is consistent with experimental data suggesting facilitating drugs
may act as a wedge to bias hERG channel equilibrium towards the open state and increase hERG current
amplitude in response to low-voltage depolarization.

Copyright © The Japanese Pharmacological Society1-B-P-016

一般演題（ポスター）

Simvastatin attenuates cardiac fibrosis during the development of chronic heart
failure via an inhibition of Hsp90-regulated cell signaling.

〇丸ノ内 徹郎、矢野 絵美、田野中 浩一
東京薬科大・薬Hsp90 is a molecular chaperone that contributes to the regulation of various cell signaling pathways. In ourprevious study, we showed that an inhibition of Hsp90 prevented cardiac fibrosis and preserved cardiac function
during the development of chronic heart failure (CHF). Simvastatin, an inhibitor of HMG-CoA reductase, hasbeen shown to inhibit the chaperone activity of Hsp90. However, it is unclear whether simvastatin can prevent
myocardial fibrosis by inhibiting Hsp90. Therefore, we examined effects of simvastatin in the models of CHF.
The results showed that treatment with simvastatin prevented cardiac fibrosis. Furthermore, simvastatin
treatment inhibited the c-Raf-Erk1/2 pathway, which contributes to cardiac fibrosis.

experiment, theinteraction of Hsp90 with c-Raf in cultured cardiac fibroblasts was decreased by the presence of simvastatin, andthe expression level of c-Raf was also reduced. Furthermore, simvastatin inhibited the fibroblast proliferationand migration. Simvastatin also attenuated collagen production by inhibiting the differentiation of cardiac
fibroblasts into myofibroblasts. These results suggest that an inhibition of Hsp90 is partially responsible for the
inhibitory effect of simvastatin on cardiac fibrosis.

Copyright © The Japanese Pharmacological Society1-B-P-017

一般演題（ポスター）

The BRG1/p300 Complex Increases the Acetylation Levels of the H3K122 in theDevelopment of Heart Failure〇船本 雅文1,2,3、砂川 陽一2,3、刀坂 泰史2,3、清水 果奈2,3、長谷川 浩二2,3、森本 達也2,3
徳島大・院医歯薬・薬理学分野、2国立病院機構京都医療セ・臨床研究セ・展開医療研、3静岡県立大・薬・分子病態

1

Epigenetic mechanisms such as histone post-translational modifications are involved in heart
failure (HF). Although the acetylation of tail domains, such as H3K9, has been extensively studied, that of
H3K122, the globular domain, has received much less attention. The H3K122 acetylation directly activatestranscription by destabilizing histone-DNA binding. However, the participation of these acetylated domains in
the development of HF remains unknown.
Phenylephrine increased the acetylation levels of H3K9 and H3K122 in Cultured
cardiomyocytes. The acetylation levels of H3K9 and H3K122 on the hypertrophic reaction gene promoters suchas BNP and b-MHC were increased in cardiomyocyte hypertrophy. In Dahl-salt sensitive rats, a heart failure
model,ChIP assays revealed that the acetylation of H3K9 on the promoters of BNP and b-MHC was

increased in left ventricular hypertrophy (LVH), while that of H3K122 was increased in HF. On the other hand,
there was no difference in the amount of transcriptional coactivator p300 recruitment in LVH and HF.
Interestingly, IP -WB showed that binding of p300 with BRG1, a key component of the SWI/SNF complex, wasenhanced in HF. The recruitment of BRG1 was increased in HF compared to LVH. Moreover, PFI-3, a BRG1
inhibitor, suppressed transverse aortic constriction-induced cardiac hypertrophy and the development of HF.
This study shows that the acetylation of H3K122 is enhanced via the interaction of p300 with BRG1
in heart failure.

Copyright © The Japanese Pharmacological Society1-B-P-018

一般演題（ポスター）

High-throughput screening for agonists of ROS production in live humanvascular endothelial cells〇笹原 智也1,2、星 美奈子1,3
1

神戸医療産業都市推進機構・先端医療研究セ・神経変性疾患研、2TAOヘルスライフファーマ・神戸研・研究開発、3京

都大・医・形態形成機構

Reactive oxygen species (ROS) are essential physiological molecules, and ROS production is strictly regulated.
In many diseases, the change of physiological function and the agonist released induce the excessive increase of
ROS production, and then cell function is impaired. In cardiovascular diseases, such as hypertension, diabetes,and Alzheimer's vascular lesions, the ROS production increases in vascular endothelial cells and eventually
disturbs the vascular function. To better understand the role of ROS in disease, it is essential to elucidate what
agonist increases the ROS production and where the ROS production occurs. Here, we show a novel highthroughput screening (HTS) protocol of what agonists induce ROS production and where it occurs in live
human vascular endothelial cells using a fluorescence probe and an image cytometer. By using this HTS
protocol, we found that Alzheimerʻs brain-derived assemblies, amylospheroids, increase the mitochondrial ROS
production because the mitochondrial ROS inhibitors (YCG-063 and mito-tempol), but not NADPH oxidase
inhibitors (VAS2870 and apocynin), blocked the ROS production. By elucidating whether ROS production
occurs in mitochondria or plasma membrane of live cells, this protocol will lead to a better understanding of therole of ROS in disease.

Copyright © The Japanese Pharmacological Society1-B-P-019

一般演題（ポスター）

Expression and function of basement membrane-derived matricryptins inthickening of mitral valve in rats

〇岡田 宗善、佐野 功汰、鈴木 竜之介、兒玉 朋子、大谷 紘資、山脇 英之
北里大・獣医

Myxomatous mitral valve disease, the most common form of canine heart disease, causes mitral regurgitation
due to valvular degeneration (proliferation and thickening of spongiosa), similar to mitral valve prolapse in
humans. The bioactive fragments cleaved from the fibrous components are called matricryptins. Arresten (α1chain) and canstatin (α2 chain), the cleaved fragments of type IV collagen, are known as basement membranederived matricryptins which are expressed in the cardiac tissue in rats. Although the expression of thesematricryptins may be altered during mitral valve degeneration, it has not been clarified. In this study, weinvestigated the expression of the matricryptins during mitral valve thickening in rats and its role in valvularinterstitial cells (VICs). The expression of canstatin was increased in the thickened mitral valve in the old rats(>36-weeks-old). Stimulation of cultured rat mitral valve with transforming growth factor (TGF)-β increased
the thickness of mitral valve and the expression of arresten. Both canstatin and arresten affected TGF-βinduced activation of VICs (increase of α-smooth muscle actin expression). These results suggest that theexpression of canstatin and arresten changes with thickening of the rat mitral valve and that they are involved in
the activation of VICs.

Copyright © The Japanese Pharmacological Society1-B-P-020

一般演題（ポスター）
2+

The regulation mechanisms of mitochondrial Ca signaling mediated by cardiac
Sigma-1 receptor〇田頭 秀章1,2、篠田 康晴2、沼田 朋大1、福永 浩司3
秋田大・院医・器官・統合生理学、2東北大・院薬・薬理学、3東北大・院薬・先進脳創薬講座

1

Cardiovascular disease (CVD) is a leading cause of death worldwide. We previously reported that the Sigma-1
receptor (Sigmar1) is down-regulated in mice with cardiac dysfunction. Recent study suggested that Sigmar1
deficient mice display cardiac dysfunction via impairment of mitochondrial function. However, the mechanismof mitochondrial quality control mediated by Sigmar1 has not been investigated in detail. In this study, weinvestigated the role of Sigmar1 for ER-mitochondrial tethering and mitochondrial Ca2+ signaling using a
Sigmar1-knockdown cardiomyocytes. We found that disruption of ER-mitochondrial tethering and reduction of
ER-mitochondrial Ca2+ transport was induced by Sigmar1 knockdown in cardiomyocytes. We also demonstrated
that Endothelin-1-induced cardiomyocyte hypertrophy is aggravated associated with induction of mitophagy inSigmar1 knockdown cardiomyocytes. These data suggest that reduction of cardiac Sigmar1 is involved inmyocyte hypertrophy by maintaining of intracellular Ca2+ signaling mediated by regulation of ER-mitochondrial
tethering.

Copyright © The Japanese Pharmacological Society1-B-P-021

一般演題（ポスター）
2+

Species difference in the intracellular Ca -mediated mechanisms in thepacemaker depolarization of mouse and guinea pig sinus node

〇尾高 椋介、濵口 正悟、行方 衣由紀、田中 光
東邦大・薬・薬物

In general, smaller animals have higher heart rates, suggesting that there are differences in the mechanisms ofpacemaker depolarization. Recently, it has been postulated that the pacemaker depolarization is influenced byintracellular Ca2+. This study intended to clarify the intracellular Ca2+-mediated mechanisms involved in thepacemaker depolarization of the mouse and guinea pig sinus node.
Microelectrode recordings revealed that the sinus node of the mouse, which had a higher beating rate, had asteeper slope of the pacemaker depolarization than that of the guinea pig. Intracellular Ca2+ interfering agents,
BAPTA and ryanodine, significantly decreased the slope in both species. In contrast, SEA0400, a specificinhibitor of the Na+-Ca2+ exchanger (NCX), as well as change to low Na+ extracellular solution, significantly
decreased the slope in the mouse, but not in the guinea pig. Confocal microscopy revealed the presence of
spontaneous Ca2+ oscillations during the interval between Ca2+ transients; Ca2+ oscillations were more
pronounced in the mouse than in the guinea pig.
These results suggested that, although intracellular Ca2+-mediated mechanisms were involved in the pacemaker
depolarization of the sinus node in both species, the NCX current was involved in the mouse but not in theguinea pig.

Copyright © The Japanese Pharmacological Society1-B-P-022

一般演題（ポスター）

Phenotypic analysis in cardiac-specific Tric-b-deficient mice

〇山崎 大樹1、松下 幸平2、徳竹 美佳1、中條 かおり1
国立医薬品食品衛研・安全性生物試験研究センター・薬理部、2国立医薬品食品衛研・安全性生物試験研究セン

1

ター・病理部

Ryanodine receptors and IP3 receptor are Ca2+ release channels which expressed on the endoplasmic reticulum(ER) membrane. When Ca2+ is released through these channels, negative charges are generated in the ER
lumen. Since the negative charge inhibits further Ca2+ release, it has been suggested that there are counter ion
channels that transport counter ions into the ER lumen to neutralize the negative charge. We have been
demonstrated that TRIC (trimeric intracellular cation) channel subtypes (A and B) played critical roles in thisprocess. TRIC-A is expressed in excitable tissues such as skeletal muscle and heart. TRIC-B is expressed
universally throughout the body. In systemic-deficient (KO) mice, type II alveolar epithelial cells are

impaired in surfactant production and secretion, resulting in impaired alveolus formation and neonatal lethality.
It has also been shown that osteogenesis imperfecta occurs due to impaired collagen secretion in osteoblasts.
Since
heart. Sinceanddouble KO mice show embryonic lethal, both subtypes may play important functions in the-KO mice are neonatally lethal, we have generated cardiac-specific-KO mice andanalyzed their function. Here, we examined the effects of β-adrenergic stimulation and cardiotoxicity on mice,
as well as the effects of drug-induced arrhythmia.

Copyright © The Japanese Pharmacological Society1-B-P-023

一般演題（ポスター）

Sex and arterial site differences in vasorelaxation via protease-activated receptor
2 in metabolic syndrome

麓 (丸山) 加菜1、懐 理紗1、McGuire John J.2、篠塚 和正1、籠田 智美1,3
1

武庫川女子大・薬・薬理2、2ウエスタン・オンタリオ大、3武庫川女子大・バイオサイエンス研

Activation of protease-activated receptor 2 (PAR2) on vascular endothelial cells causes vasorelaxation. Nitricoxide (NO)-mediated vasorelaxation of the aorta is impaired in male SHRSP.Z-/IzmDmcr (SP.ZF) rats

with metabolic syndrome (MetS), but PAR2-mediated vasorelaxation is preserved. In the current study, weinvestigated whether PAR2-mediated vasorelaxation in the thoracic aorta and superior mesenteric artery (MA)differed by sex and arterial site in SP.ZF rats, and SHR.Cg-/NDmcr (CP) rats at 23‒26 weeks of age, twodifferent models of MetS.
Vasorelaxation was examined using the organ bath method. In isolated aortas from SP.ZF rats, vasorelaxations
evoked by 2fLIGRLO, a PAR2 agonist, and by acetylcholine (ACh) were greater in females than in males, but inthe case of CP rats, only ACh-induced relaxation was greater in females. In MA, 2fLIGRLO-induced
vasorelaxation was smaller in females than in males in both strains. However, ACh-induced relaxations did not
differ between sexes. Nitroprusside-induced relaxation did not differ significantly between sexes or arterial sites.
The findings demonstrate the presence of sex- and arterial site-dependent differences in PAR2-mediatedvasorelaxations in MetS. The results indicate less PAR2-mediated relaxation in MAs of female rats under thecondition of a presumably maintained NO-mediated vasorelaxation pathway. Further studies are needed toelucidate the pathophysiological significance of lower capacity for vasorelaxation via PAR2 in females with MetS.

Copyright © The Japanese Pharmacological Society1-B-P-024

一般演題（ポスター）

Analysis of vasoconstrictor effects of local anesthesia, mepivacaine,
via α receptors and V1Areceptor.

〇池田 哲朗1、齋藤 良介1,2、田中 夏幹1、益見 厚子2
青森大・薬・病態分子薬理学、2青森大・薬・分子薬理学

1

Several studies suggested that the amide-type local anesthetic mepivacaine has a longer duration of local
anesthetic effect than the same amide-type lidocaine, and has a vasoconstrictor action. However, the mechanismof action on α2 and α1 adrenoceptors involved in peripheral vasoconstriction remains unclear.
0.25% mepivacaine with various ligands such as α adorenoceptors antagonists is randomly and blindly injected
intracutaneously at a dose of 0.1mL into the back of shaved Hartly male guinea pigs (weight 300-350g). After
marking the area around the wheal and confirming that there is a normal skin contraction reaction outside thewheal, six pricks were aplied inside each wheal. The test of six pinpricks was applied at intervals of about 3 to 5
seconds every 5 minutes. The number of times that did not respond to stimulus was measured and the sum(maximum value:132) served as an anesthetic score which indicates the degree of local anesthesia.
Mepivacaine dose-dependently prolonged the duration of anesthesia and increased local anesthesia scores.
Therefore, 0.25% mepivacaine was used and compared with 0.25% lidocaine and 0.25% xylocaine. The
anesthetic duration of 0.25% mepivacaine was longer than that of lidocaine and shorter than that of xylocaine.
0.25% mepivacaine were combined with 1 μM yohimbine, 1 μM prazosin, 1 μM JP1302, 10 μM BRL44408,
1 μM SR49059 (selective V1A receptor blocker V:vasopressin), 10 μM indoramin, 0.5 μM BMY7378, 5 μM
cyclazosin, 1 nM silodosin respectively. After mixing, the effect of various antagosits on mepivacaine were
examined. As a result, antagonists other than 10 μM BRL44408 and 1 nM silodosin decreased the duration ofmepivacaine anesthesia. These results indicated that mepivacaine's peripheral vasoconstrictor activity is
mediated at least by α2C, α1B, α1D and V1Areceptors.

Copyright © The Japanese Pharmacological Society1-B-P-025

一般演題（ポスター）

Do female mice acquire a preference for the unattractive male mouseencountered after cocaine administration?

〇大西 克典、河原 幸江、大西 陽子、西 昭徳
久留米大・医

Occasional incidents of drug addiction among celebrities have been reported, and sometimes the presence ofthe opposite sex flickers. Even trafficking of women sometimes involves the use of illegal drugs. Whether drugs
induce not only drug dependence but also an associated preference for the opposite sex is an important questionin understanding and solving such cases.
Previously, we have established a female male preference test (FMPT), in which four male mice are compared to
distinguish between attractive and unattractive male mice. Using this established system, we examined whether
conditioning female mice with an addictive drug would increase their preference for the unattractive male
mouse that was with them at the time.
Briefly, the female mice were administered drugs (cocaine 7.5 mg/kg ip or morphine 15 mg/kg sci) and thenallowed to meet an unattractive male mouse defined by FMPT for 15 minutes for 3 consecutive days, and were
examined by FMPT on the fourth day.
In the results, morphine treated female mice showed significant higher preference to the same male mouse as
saline treated control female mice, and the mouse wasnʻt the conditioned unattractive male mouse, moreover,cocaine treated female mice didnʻt show significantly different preference to specific male mice including theconditioned unattractive male mouse, indicating that those drugs could have no effect to increase the preference
to the conditioned unattractive male mouse.
However, since sexual activity might also be necessary for the attraction, the effects of sexual activity undercocaine administration into female mice will also be examined and reported.

Copyright © The Japanese Pharmacological Society1-B-P-026

一般演題（ポスター）

Molecular mechanism of schizophrenia-like behavioral deficits induced bymethylglyoxal detoxification impairments〇鳥海 和也1、小池 伸2、段 孝3、鈴木 一浩1,4、宮下 光弘1,5、堀内 泰江1、小笠原 裕樹2、宮田 敏男3、糸川 昌成1、新井 誠1
1

都医学研・精神行動医学・統失PJ、2明薬大・薬・分析化学、3東北大・院医・分子治療学、4信州大・医・精神医学、5都医

学研・社会健康医学

Methylglyoxal (MG) is a reactive and cytotoxic alpha-dicarbonyl product of glycolysis. Various detoxification
systems work together

to remove highly toxic MGs, including a glyoxalase system by Glyoxalase 1(GLO1) and GLO2, and the scavenging system by vitamin B6 (VB6). We found that VB6 levels in peripheral
blood of the schizophrenia patients with GLO1 dysfunction are significantly lower than that of healthy controls.
However, the effects of the MG detoxification deficits on the pathophysiology of schizophrenia remains poorly
understood. Here, we generated a new mouse model of schizophrenia with impaired MG detoxification by
feedingknockout mice VB6-deficent diets (KO/VB6(-)). KO/VB6(-) mice accumulated MG in theprefrontal cortex (PFC), hippocampus, and striatum, and displayed behavioral deficits, such as prepulse
inhibition (PPI) deficit. Furthermore, we demonstrated abnormal mitochondrial respiratory function andsubsequently enhanced oxidative stress in the PFC of KO/VB6(-) mice in the PFC. Finally, administration of anantioxidant resveratrol improved PPI impairment as well as oxidative stress. These results suggest that the
combination of GLO1 dysfunction and VB6 deficiency results in mitochondrial dysfunction and increasedoxidative stress in PFC, resulting in schizophrenia-like behavioral disorders.

Copyright © The Japanese Pharmacological Society1-B-P-027

一般演題（ポスター）

Axo-axonic cells in the amygdala regulate associative fear learning.

〇中嶋 美紀1、池谷 裕二1,2、森川 勝太1,2
1

東京大・院薬・薬品作用、2東京大・Beyond AI研究推進機構

Activity and plasticity of excitatory neurons are tightly regulated by local inhibitory neurons in a spatiotemporal
specific manner. Axo-axonic cells (AACs) are a unique type of inhibitory neurons that express parvalbumin andinnervate the axon initial segment (AIS) of excitatory neurons. While their anatomical features have been
identified, the genetical marker and functional role remain unclear. Here, we show that vasoactive intestinal
peptide receptor 2 (Vipr2)-expressing inhibitory neurons in the basolateral amygdala (BLA) exhibit theanatomical and electrophysiological properties typical of AACs. Furthermore, using an AACs-specific labelingapproach, we conducted in vivo fiber photometry recording and functional inhibition experiments in a cued fear
conditioning test. The activity of AACs was increased for both conditioned stimulus (CS: tone) andunconditioned stimulus (US: foot-shock), and inhibition of GABA transmission of AACs impaired cued fear
conditioning. These results suggest that AACs are important for memory acquisition. Finally, we employedprojection-specific monosynaptic rabies virus tracing to identify the direct monosynaptic inputs cells to AACs in
the BLA. This study provides new insights into the detailed functional roles of AACs in the BLA.

Copyright © The Japanese Pharmacological Society1-B-P-028

一般演題（ポスター）

Blockade of D-serine signaling and adult hippocampal neurogenesis attenuates
remote contextual fear memory〇森 寿1,2,3、倪 献策1,2、井上 蘭1,3
1

富山大・院医薬、2富山大・大学院生命融合科学教育部・分子神経科学講座、3富山大・アイドリング脳科学研究セン

ター

Memory retrieval can trigger destabilization followed by reconsolidation for maintaining or enhancing original
fear memory. Therefore, blockade of reconsolidation could weaken the original fear memory. The -methyl-Daspartate (NMDA) receptor and hippocampal neurogenesis play crucial roles in hippocampus-dependent
memory processes, including reconsolidation. In this study, first, NMDA receptor signaling was downregulatedby the genetic reduction of its co-agonist, D-serine, and the neurogenesis was ablated by focal X-ray irradiationon the hippocampus. We found that a progressive decrease in freezing following each retrieval, leading to an
attenuation of remote contextual fear memory on day 28. Second, after conditioning, pharmacologicalapproaches to simultaneously block D-serine signaling and inhibit neurogenesis, resulting in a similar
suppressive effect on the remote fear memory. The present findings provide insights into the role of D-serinemediated NMDA receptor signaling and neurogenesis in memory retrieval and the maintenance of remote fear
memory, and provide a new strategy to improve exposure-based therapy for post-traumatic stress disorder(PTSD) treatment.

Copyright © The Japanese Pharmacological Society1-B-P-029

一般演題（ポスター）

Enhancement neuronal activity in the basolateral amygdala in mice with thepreference of nicotine intake〇藤井 拓磨、泉尾 直孝、浅野 昴志、新田 淳美
富山大・院医薬・薬物治療学研究室[Introduction] Despite the health damage risks of tobacco, success rate of smoking cessation is quite low.
Tobacco dependence has aspects of substance and behavioral addiction. The latter aspect is supported by thelimited efficacy of smoking cessation by nicotine replacement therapy such as nicotine gum or patches. In thisstudy, to clarify the mechanism of behavioral addiction with smoking, we established the model of nicotine
intake behaviors in mice corresponding to behavioral addition in human smokers, and examined the brain
region associated with nicotine intake behaviors. [Methods] Two drinking bottles were placed on the housing
cage of mice (C57BL/6J, 8-9 weeks, male). After the habituation period, mice were exposed to the watercontaining nicotine (75µg/ml) filled in both bottles for one week to learn nicotine intake behavior (behavioral
priming). Then, to measure the preference to nicotinic solution, bottles containing vehicle or nicotinic solution
were presented to mice. [Results] Mice which experienced behavioral priming much more preferred to thenicotinic solution than those without behavioral priming. Immunohistochemistry to FosB, a marker of neuronal
activity, revealed that larger number of FosB-positive neurons were observed in the basolateral amygdala (BLA)in mice without the preference to the nicotine solution, than those with preference. [Conclusion] Enhanced
neuronal activity in the BLA is suggested to be associated with behavioral addiction of tobacco dependence.

Copyright © The Japanese Pharmacological Society1-B-P-030

一般演題（ポスター）

Evaluation of the circadian rhythms of activity in mice implanted with humanstomach cancer cell lines.

〇上野 晋1、後藤 元秀1、野中 美希2、丸山 崇3、石塚 恒年1、長谷川 渉1、溝上 峻1、上田 陽一3、上園 保仁21

産業医大・産生研・職業性中毒、2慈恵医大・医・疼痛制御研究、3産業医大・医・第１生理Objective: Circadian rhythm is endogenous 24-hr oscillations usually entrained to the daily environmental cycleof 12/12 h light/dark (LD). It has been reported that disruption of circadian rhythm causes delirium, which isalso often developed in patients with advanced cancer. However, the mechanisms by which the progression ofcancer disrupts circadian rhythms and ultimately leads to the development of delirium has not been yet
understood. We have recently established cancer-induced cachexia model mice by implantation of humanstomach cancer cell line 85As2. In this study, we investigated the circadian rhythms of locomotor activity in
85As2-implanted mice.
Methods: 85As2 cells were implanted into 8-week-old male BALB/c nude mice (2x105 cells/mouse). Home-cage
activity was measured after 2 weeks from implantation under LD. Mice were then placed in constant darkness(DD) or light (LL), and finally returned to LD cycle. The day-night variation were analyzed using Actogram.
Results: The active phase of 85As2-implanted mice gradually shifted from dark to light between 2 to 4 weeks
after implantation, which was almost reversed after 4 weeks. The free-running period under DD or LL
condition was significantly shorter compared to the control. Even after returning to LD, the active phaseremained reversed in 85As2-implanted mice.
Summary: 85As2-implanted mice demonstrated the reverse of day-night variation without synchronization ofphotoperiod, and shortened free-running period, suggesting that 85As2-implantation may directly affect thecircadian rhythm formation.

Copyright © The Japanese Pharmacological Society1-B-P-031

一般演題（ポスター）

Maternal obesity impairs social behavior in female offspring by decreasing
central estrogen function〇鎌田 知紘、植田 大暉、米持 奈央美、池田 弘子
星薬科大・薬・薬物治療

Maternal obesity is reported to increase risk for metabolic diseases in offspring. Furthermore, recent researchsuggests that maternal obesity also affects the central nervous system in offspring, but a mechanism is unclear.
The present study examined the effects of maternal obesity on central nervous system function in offspring.
Female 8-week-old C57BL/6J mice were fed with a high-fat diet (HFD) or a regular diet (RD) for 4 weeks andmated with male 8-week-old C57BL/6J mice. The experiments were conducted when offspring reached 8 weeks
of age. In the social interaction test, interaction time with a stranger mouse in female offspring of mice fed with
HFD was reduced compared to female offspring of mice fed with RD; no such effect was observed in maleoffspring. These results suggest that only female offspring of obese mice show impairment in social behavior.
The mRNA levels of estrogen receptor alpha in the amygdala, which regulates social behavior, were reduced infemale offspring of mice fed with HFD, while plasma estradiol levels were unchanged. In addition, the estrogen
receptor antagonist fulvestrant reduced interaction time with a stranger mouse. Taken together, our results
suggest that maternal obesity impairs social behavior in female offspring by decreasing central estrogen
function.

Copyright © The Japanese Pharmacological Society1-B-P-032

一般演題（ポスター）

The role of sign tracking in the formation of process addiction〇太田 宏之、石塚 俊晶
防衛医科大・医学科・薬理Current treatment for process addiction, such as gambling disorder and game addiction, is primarily
psychotherapy, and effective pharmacotherapy has not been established. One reason is the lack of appropriate
behavioral test batteries for mice to identify the causes of addiction and conduct pharmacological tests. The
scarcity of high-value rewards is generally considered the cause of addiction, and experimental behavioral tasks
have been proposed accordingly. In recent years, however, it has become clear that signs that foretell rewards are
involved in the formation of gambling addiction. In the present study, we developed a 2-hole nose poke-selective
operant task for mice in which two retractable levers were used as predictive signs for the reward. As a result,mice showed adherence to the option with predictable rather than unpredictable reward signs. Adherence to thepredictable reward signs continued even after the reward probability decreased. We also found that the
suboptimal behavior formation was correlated with the frequency of sign tracking. This study allows us to study
the role of sign tracking in the shape of process addiction using genetically engineered mice and drug
administration models.

Copyright © The Japanese Pharmacological Society1-B-P-033

一般演題（ポスター）

Reactivation of hippocampal place cells after experiencing multiple
environments〇横井 太紀1、鹿野 悠2、柳下 晴也2,3、池谷 裕二2,4、佐々木 拓哉2,3
1

東北大・薬・薬理、2東京大・院薬・薬品作用、3東北大・院薬・薬理、4Beyond AI 研究所

The hippocampus plays important roles in learning and memory. In the hippocampus, spatial information is
encoded by place cells during an experience and consolidation of spatial memory is supported by reactivation ofplace cells during rest/sleep periods. It remains to be unknown how multiple spatial experiences that are
encoded by a subset of place cell ensembles are reactivated in subsequent rest/sleep periods. To address thisissue, we recorded spike patterns of hippocampal CA1 and CA3 place cells in rats that sequentially experienced
five different rooms. We confirmed that CA1 cells had larger numbers of place fields defined from individualrooms that CA3 cells, suggesting sparser spatial representations by CA3 cells. Overall, increases in reactivation
rates of CA1 place cells from pre-rest to post-rest periods were correlated with the numbers of place fields,suggesting stronger reactivation in neurons that encoded more information during awake periods. We now
analyze synchronous spike patterns of multiple neurons to clarify how experiencing multiple environments are
encoded and consolidated by hippocampal neuronal ensembles.

Copyright © The Japanese Pharmacological Society1-B-P-034

一般演題（ポスター）

Effect of μ-opioid receptor selective antagonist β-funaltrexamine on
spontaneous behavior in mice

〇北中 順惠1、北中 純一2、富田 和男3、五十嵐 健人3、田中 康一2、西山 信好2、佐藤 友昭31

兵庫医科大・医、2兵庫医科大・薬・薬理、3鹿児島大・院医歯・応用薬理In this presentation, we hypothesized that μ-opioid receptor antagonists could inhibit the natural reward aswell as drug addiction such as opioids and stimulants. To evaluate natural reward, horizontal running wheels
were applied to mice administered with a μ-opioid receptor antagonist β-funaltrexamine (β-FNA). In
addition to the rotation of running wheels, horizontal locomotion, amounts of food intake and drinking, andaccess of food container were measured simultaneously by using a multi-configuration apparatus (0700/1900
light on/off). In naïve mice, horizontal locomotion increased day by day for three testing days. The increment
was parallel to the increase in the rotation of running wheels but not to the number of accesses to foodcontainers. The increase in the rotation of running wheels was sensitized, and the amount of food intake wasunchanged in naïve mice. In mice group which was administered with a single injection of 5 mg/kg β-FNA, thelocomotion and thus the rotation of the running wheels decreased especially in the first testing day. In contrast,
no significant behavioral change was observed in mice under a continuous s.c. administration with osmotic
mini-pumps (3.5 mg /kg of β-FNA for three days (71.5 h)). These results indicate that a transient increase in
the levels of β-FNA reduced motivative behaviors in mice.

Copyright © The Japanese Pharmacological Society1-B-P-035

一般演題（ポスター）

Pretreatment with CHIR-99021, a GSK-3 inhibitor, partially attenuates
methamphetamine-induced stereotyped behavior in mice

〇北中 純一1、北中 順惠2、富田 和男3、五十嵐 健人3、田中 康一1、佐藤 友昭3、西山 信好1
1

兵庫医科大・薬・薬理、2兵庫医科大・医・薬理、3鹿児島大・院医歯・応用薬理In mammals, glycogen synthase kinase-3 (GSK-3) is formed as the two isoforms termed GSK-3α and GSK-3β.
GSK-3β is present in a high concentration in the abundance of tissues in the central nervous system, regulatinga crucial role in neuronal signaling pathways. The research for involvement of GSK-3β signaling in drug abuseliability has been progressed based on the studies investigating molecular and cellular mechanism of action, but
few reports have been made on animal research so far. In this presentation, we demonstrate that pretreatment ofmice with CHIR-99021 (5, 10, 15 mg/kg, s.c.), a GSK-3 inhibitor, attenuated methamphetamine (METH)induced stereotyped behavior (10 mg/kg of METH) in a dose-dependent fashion. Maximal inhibitory effect (ca.
50%) was observed at 10 and 15 mg/kg of CHIR-99021. CHIR-99021 dose-dependently attenuated theexpression frequency of METH-induced stereotyped biting whereas CHIR-99021 increased the expression
frequency of persistent locomotion. These observations are different results from those of other GSK-3
inhibitors such as SB216763 and AR-A014418, suggesting a possibility of CHIR-99021-specific effect of METH
actionCopyright © The Japanese Pharmacological Society1-B-P-036

一般演題（ポスター）

Protective Effect of Bcl-2-associated athanogene (BAG) 3 in Mouse
Neuroblastoma N1E115 Cells〇東尾 里英子、猪俣 結衣、高橋 晋太郎、玉田 さち、夏堀 陽子、三部 篤
岩手医科大・薬・薬剤治療

Bcl-2-associated athanogene (BAG) 3 is known as a regulator of cell death as well as autophagic protein
turnover in the heart. However, little information is present in functional role of BAG3 in neurons. We analyzed
the functional role of BAG3 in N1E115 cells derived from mouse neuroblastoma. While overexpression of BclxL, a member of anti-apoptotic factor, can prevent cell death against 30 nM staurosporine, no obvious protective
effect was seen by overexpression of BAG3 in N1E115 cells. In contrast, combined overexpression of BAG3 with
Bcl-xL can enhanced the effect by overexpression of Bcl-xL in N1E115 cells. Slight protective effect of BAG3
against ABT263, Bcl-2 inhibitor, was also observed in N1E115 cells. To address underlying mechanisms ofprotective effect of BAG3, overexpression as well as knockdown of BAG3 was performed. Although knockdown
of BAG3 in N1E115 cells using si-RNA specifically targeted to BAG3 enhanced mitochondrial Bak1 protein
levels, no alteration in Bak1 gene expression, and gene expression and protein levels of Bax, Bcl-2 was observed.
Concomitant with increased Bak1, the proportion of TUNEL-positive N1E115 cells was increased, andoverexpression of BAG3 led to suppression of the Bak1 level. Bafilomycin A1, an autophagy inhibitor, caninhibit the modification of Bak1 protein level by BAG3. These results suggest that BAG3 may be critical toprotein level in Bak1 via modification of autophagy activity, and that autophagy regulation by BAG3 may play animportant role in the apoptosis of neuronal cells.

Copyright © The Japanese Pharmacological Society1-B-P-037

一般演題（ポスター）

The effect of a miR-96-5p inhibitor delivery to brain using microbubbles andultrasound technology on neuroprotection and autophagy activation〇木下 千智1、青山 晃治1、鈴木 亮2、松村 暢子1、小俣 大樹2、丸山 一雄2、中木 敏夫31

帝京大・医・薬理、2帝京大・薬・薬物送達、3帝京大・医療技術・柔道整復

Glutathione (GSH) is an important antioxidant that plays a critical role in neuroprotection. GSH depletion inneuron induces oxidative stress promoting neuronal damage, which is regarded as a hallmark of the early stage
in some neurodegenerative diseases, such as Alzheimerʻs disease and Parkinsonʻs disease. The neuronal GSH
levels are mainly regulated by excitatory amino acid carrier 1 (EAAC1) and its inhibitory protein, glutamate
transporter-associated protein 3-18 (GTRAP3-18). In this study, we found that GTRAP3-18 levels wereincreased by the up-regulation of the microRNA miR-96-5p, which has been reported to decrease EAAC1 levels
in our previous study. We also discovered that neuro-oncological ventral antigen 1 (NOVA1) is an intermediate
protein for GTRAP3-18 expression via miR-96-5p. Moreover, we show that the intra arterial administration of amiR-96-5p-inhibiting nucleic acid to living mice by a drug delivery system using microbubbles and ultrasounddecreased the levels of GTRAP3-18 via NOVA1, while increased the levels of both EAAC1 and GSH in themouse brain. Moreover, we found that the administration of a miR-96-5p inhibitor increased autophagy
activation in the mouse hippocampus. These findings suggest that the delivery of a miR-96-5p inhibitor to thebrain would efficiently increase the neuroprotective activity by increasing GSH levels via EAAC1, GTRAP3-18
and NOVA1.

Copyright © The Japanese Pharmacological Society1-B-P-038

一般演題（ポスター）

3‘,4‘,7-Trihydroxyflavone inhibits NO production in LPS-activated MG6 microglial
cells by suppressing the JNK–STAT1 pathway〇赤石 樹泰、山本 昇平、阿部 和穂
武蔵野大・薬We investigated the effects of the natural flavonoid 3ʻ,4ʻ,7-trihydroxyflavone on lipopolysaccharide (LPS)induced neuroinflammatory responses in MG6 microglial cells. 3ʻ,4ʻ,7-Trihydroxyflavone inhibited LPS-induced
nitric oxide (NO) production and the upregulation of inducible NO synthase (iNOS) in MG6 cells. 3ʻ,4ʻ,7Trihydroxyflavone also suppressed LPS-induced phosphorylation of signal transducer and activator oftranscription 1 (STAT1), which is crucial for iNOS expression. LPS stimulation induced rapid phosphorylationof c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), and extracellular signalregulated kinase (ERK) in MG6 cells. 3ʻ,4ʻ,7-Trihydroxyflavone significantly inhibited the LPS-induced
phosphorylation of JNK, but not that of ERK and p38 MAPK. The inhibitory effect of 3ʻ,4ʻ,7-trihydroxyflavone
on NO production was mimicked by pharmacological inhibition of the JNK signaling pathway with SP600125.
Furthermore, SP600125 significantly inhibited LPS-induced phosphorylation of STAT1 in MG6 cells. These
results suggest that 3ʻ,4ʻ,7-trihydroxyflavone exerts anti-neuroinflammatory effects via inhibition of the JNKSTAT1 pathway in microglia.

Copyright © The Japanese Pharmacological Society1-B-P-039

一般演題（ポスター）

Brain Transport of Non-Esterified Docosahexaenoic Acid Across the Blood–Brain
Barrier is Decreased in Middle-Aged and Aged Mice

〇岩尾 卓朗、高田 芙友子、片岡 泰文、横谷 みき、有留 尚孝、安永 美保、道具 伸也
福岡大・薬・応用薬剤学教室

Nutrients are actively taken up by the brain through various transporters at the blood‒brain barrier (BBB).
Orally supplied DHA must be transported from the circulating blood to the brain across the BBB throughtransport carriers including major facilitator superfamily domain-containing protein 2a (MFSD2A) for esterified
DHA and fatty acid-binding protein 5 (FABP5) for non-esterified DHA. Although BBB integrity is known to bealtered in aging, the impact of aging on DHA transport across the BBB is not well understood. Here, we used 2-,8-, 12-, and 24-month-old male C57BL/6 mice to evaluate the brain uptake of [14C]DHA, as non-esterified
form, using an in situ transcardiac perfusion technique. Primary culture of rat brain endothelial cells (RBECs)
were used to evaluate the effect of siRNA-mediated MFSD2A knockdown on cellular uptake of [14C]DHA. We
found that brain uptake of [14C]DHA was inhibited by an excess amount of unlabeled DHA in 2-month-old
mice. Transfection of MFSD2A siRNA into RBECs decreased the protein expression levels of MFSD2A andcellular uptake of [14C]DHA. 12- and 24-month-old mice showed significant decrease in brain uptake of [14C]
DHA and MFSD2A protein expression in the brain microvasculature compared with 2-month-old mice,nevertheless FABP5 protein expression was up-regulated with age. Our findings suggested that MFSD2A isinvolved in non-esterified DHA transport at the BBB. The middle-aged and aged brain has decreased DHA
transport across the BBB due to age-related down-regulation of MFSD2A rather than FABP5.

Copyright © The Japanese Pharmacological Society1-B-P-040

一般演題（ポスター）

Neurotoxicity assessment of pyrethroids using multielectrode array recordingsof human iPSC-derived neurons〇安彦 行人、山田 茂、諫田 泰成
国立衛研・薬理部

Neurotoxicity of environmental chemicals, such as pesticides, is a worldwide concern in human welfare. It ischallenging to identify human neurotoxicity by animal tests, due to differences in species, costs, and labor.
Human induced pluripotent stem cells (hiPSC)-based in vitro tests might be a valuable method to evaluate
chemical neurotoxicity, but the standard procedure, such as cell lines and endpoints, has not been fully
understood.
In this study, we examined the neurotoxicity of pyrethroid insecticides using the multielectrode array (MEA)
recordings of hiPSC-derived neurons. We exposed the hiPSC-derived neurons XCL-1 to pyrethroid insecticides
and performed MEA recordings using MED64-Presto.
Exposure with pyrethroids to hiPSC-derived neurons reduced neural network function, such as spikes per
network burst and network burst duration in a dose-dependent manner. RT-PCR analysis revealed theexpression of the pyrethroid-sensitive voltage-gated sodium channels (VGSCs) in hiPSC-derived neurons. The
half-maximal inhibitory concentrations (IC50s) of the MEA parameters were lower than the 30% effective
dosages (ED30s) of motor activity repression obtained from the animal experiments.
Taken together, MEA recordings in network activity of hiPSC-derived neurons could be an effective tool toscreen compounds with neurotoxicity.

Copyright © The Japanese Pharmacological Society1-B-P-041

一般演題（ポスター）

Low concentration of methotrexate causes toxicity against neuron during
synapse formation〇山村 真伊、花村 健次、川辺 浩志
群馬大・院医・薬理High-dose methotrexate (MTX) application is widely used for the treatment of acute lymphoblastic leukemia inchildren. MTX can cross blood-brain barrier to reach neurons when applied with a high dose. Indeed, high-dose
MTX application could cause neurological side effects including cognitive impairment. In our previous reports,
we established a high-throughput imaging system for the detection of the effects of various neurotoxic
compounds using primary cultured neurons. Using this system, we studied the neurotoxic effects of various
concentrations of MTX in the present study. For this purpose, primary cultured cerebral cortical cells wereprepared from embryonic days 17 Wister rat. Cerebral cortical cells were cultured for 21 days on 96-well
microplates. At 4 days in vitro (4 DIV), MTX was applied to the cultured cells in each well at the concentrationof 0.01, 0.03, 0.1, 0.3, 1, 10, or 100 µM, respectively. At 21 DIV, cerebral cortical cells were fixed withparaformaldehyde and stained with DAPI and with neuronal and synaptic markers. To our surprise, lowconcentrations of MTX reduce the number of cultured cerebral cortical neurons in the well. Our finding
indicates that MTX could be toxic at a concentration lower than what we expected.

Copyright © The Japanese Pharmacological Society1-B-P-042

一般演題（ポスター）

The effect of opioid promoting social behavior is negatively regulated by theactivity of dorsomedial part of periaqueductal grey

〇大波 壮一郎1,2、山川 英訓1,2、大石 直也2、小川 公一1,2
1

塩野義製薬・創薬疾患研究所、2京都大・医・メディカルイノベーションセンター・SKプロジェクト

The opioid system has been reported to play a crucial role in modulating social behavior in humans and animals.
However, brain regions which mediate the effect of opioids on promoting sociability and reason why the effect issuppressed depending on the dose are not determined yet. Opioid receptors are densely distributed in mPFC
and NAc, which could promote sociability. On the other hand, PAG, a key region for analgesia of opioids, isreported to negatively affect sociability. Thus we hypothesized that sociability would be promoted by controllingthese regions in a well-balanced manner. Firstly, we confirmed that morphine (Mor, s.c.) with a low dose
increased social interaction behavior in mice. At that dose, the number of c-Fos-positive cells was significantly
increased in mPFC and NAc. At a higher dose, dorsomedial (dm), but not dorsolateral/lateral, PAG was
activated in addition to mPFC and NAc. At last, we found that the effect on social behavior induced by Mor (lowdose, s.c.) was antagonized by topical administration of high dose Mor to dmPAG. These results suggest that thedmPAG works as a gate of social behavior promoted by Mor-induced activation of mPFC and NAc. A new
opioid drug with an appropriate action that does not activate dmPAG may be a new therapeutic strategy for the
deficit of sociability in mental disorders.

Copyright © The Japanese Pharmacological Society1-B-P-043

一般演題（ポスター）

Pyrylium based derivatization imaging mass spectrometer revealed the
localization of L-DOPA〇平 修1、鹿野 仁美1、池田 明夏里2、寺内 勉2、横山 順2
1

福島大学・農学群・食農学類、2大陽日酸（株）・SI事業部

Simultaneous imaging of l-dihydroxyphenylalanine (l-DOPA), dopamine (DA) and norepinephrine (NE) in thecatecholamine metabolic pathway is particularly useful because l-DOPA is a neurophysiologically important
metabolic intermediate.
In this study, we found that 2,4,6-trimethylpyrillium tetrafluoroborate (TMPy) can selectively and efficiently
react with target catecholamine molecules. Specifically, simultaneous visualization of DA and NE as metabolites
of l-DOPA with high steric hinderance was achieved by derivatized-imaging mass spectrometry (IMS).
Interestingly, l-DOPA showed strong localization in the brainstem, in contrast to the pattern of DA and NE,
which co-localized with tyrosine hydroxylase (TH).
In addition, to identify whether the detected molecules were endogenous or exogenous l-DOPA, mice were
injected with l-DOPA deuterated in three positions (D3-l-DOPA), which was identifiable by a mass shift of 3Da.
TMPy-labeled l-DOPA, DA and NE were detected at302.1, 258.1 and 274.1, while their D3 versions weredetected at 305.0, 261.1 and 277.1 in mouse brain, respectively. l-DOPA and D3-l-DOPA were localized in the
BS. DA and NE, and D3-DA and D3-NE, all of which are metabolites of L-DOPA and D3-l-DOPA, werelocalized in the striatum (STR) and locus coeruleus (LC). These findings suggest a mechanism in the brainstemthat allows l-DOPA to accumulate without being metabolized to monoamines downstream of the metabolic
pathway.

Copyright © The Japanese Pharmacological Society1-B-P-044

一般演題（ポスター）

High-content analysis using drebrin immunocytochemical images of cultured rat
hippocampal neurons〇間瀬 省吾1,2、光岡 俊成3、小金澤 紀子2、山崎 博幸2,4、加藤 祐一1、筒井 泉雄1、川辺 浩志2、白尾 智明2、関野 祐子
11

東京大・院農学生命科学、2群馬大・院医、3北海道科学大・薬、4群馬医療福祉大・社会福祉

Drebrin is a major F-actin binding protein in dendritic spines that is critically involved in their morphologicalplasticity. Subcellular localization of drebrin is dependent on NMDAR activity. Drebrin's change in dendritic
spines is an indicator of toxic effect of compounds to the brain and can be used for prediction of brain
dysfunction prior to the neuronal cell death. In the present study, we have developed new drebrin clusters
analysis method applied the confocal high-content screening method. Hippocampal neurons prepared fromembryonic rats (SKY neuron, AlzMed, Inc., Tokyo) were incubated in 96-well microplates. After 21 days, thecultured neurons were treated with 10, 100 µM glutamate. After the treatments, they were fixed and processed
for immunocytochemistry to visualize drebrin, MAP2 and cell nucleus. After automated image acquisitions,
neuron number, dendrite length, drebrin clusters were quantified with original algorithm. The high-throughput
immunocytochemical assay demonstrated that glutamate treatment decreased drebrin cluster density. The ratio
of decrease in the density was concentration dependent and found to be dependent on the number ofsurrounding neurons. Our method is sensitive enough to detect the interaction with NMDAR and is useful for
drug screening studies for synaptic dysfunction.

Copyright © The Japanese Pharmacological Society1-B-P-045

一般演題（ポスター）

Single-cell analysis of electrical activities in human iPS cell-derived neuralnetworks using 236,880-electrode CMOS-MEA〇松田 直毅、韓 笑波、鈴木 郁郎
東北工業大

In vitro microelectrode array (MEA) assessment using human induced pluripotent stem cell (iPSC)-derived
neurons holds promise as a method of seizure and toxicity evaluation. However it is difficult to detect the
response of drugs with different mechanisms of action with a single parameter, and the analysis method has
become an issue. One effective way to solve this problem is to obtain more detailed information on neuralnetwork activity. Therefore, in this study, we cultured human iPS cell-derived cortical neurons on a 236,880electrode CMOS-MEA and obtained precise single-neuron electrical activity.
As a result of acquiring spontaneous activity after 6 weeks of culture, an average of 296 ± 47 neurons (n = 9
wells) were identified, and network bursting was observed. There, 4-AP 10-30 µM, PTX 1-10 µM, AP5 25 µM,
and CNQX 30 µM were administered, and in addition to conventional network burst analysis, we analyzed theburst activity of single-neuron and the synaptic connections between neurons that form networks. As a result of
burst analysis, parameters with statistically significant changes were different between network burst and single
burst, and the number of concentrations with significant differences also increased with single burst. In
addition, in synaptic connections analysis, 4-AP showed no significant change, whereas PTX showed enhanced
synaptic connections. CMOS-MEA, which can accurately acquire the electrical activity of single neurons, canincrease the number of parameters that can be used to evaluate the effects of drugs, so it is effective as a method
for predicting toxicity and the mechanism of action of compounds.

Copyright © The Japanese Pharmacological Society1-B-P-330

一般演題（ポスター）

Neuroprotective Effect of an Inhibitor of Hypoxia-inducible Factor-Prolyl
Hydroxylase in a Cell Culture Model of Parkinson‘s disease〇藤牧 綾香、大内 一輝、村上 貴規、滝沢 進之佑、栗田 尚佳、保住 功、位田 雅俊
岐阜薬科大・薬・薬物治療学研究室

Oxidative stress is associated with the progression of neurodegenerative diseases such as Parkinsonʻs disease(PD). In the present study, to examine whether a hypoxia-inducible factor (HIF)-prolyl hydroxylase inhibitor
has neuroprotective effect against α-synuclein (syn)-induced neurotoxicity in a cellular model, we used FG-4592, also known as roxadustat. To investigate the effect of FG-4592 against, we evaluated the α-syn protein
level. mRNA levels of oxidative stress response genes whose transcription was regulated by HIF-1α wereanalyzed. Immunofluorescence staining with redox-sensitive dyes was also performed to examine the α-syninduced oxidative stress. Previously, we succeeded in generating a new α-syn stably expressing cell line. In thiscell line, we found that oxidative stress induced by α-syn caused cell death. FG-4592 exhibited significant
neuroprotective effects against α-syn-related neurotoxicity. However, FG-4592 did not affect α-syn protein
levels. FG-4592 induced heme oxygenase-1 (HO-1) expression levels in a concentration-dependent manner,suggesting that FG-4592 reduced oxidative stresses

the induction of HO-1. Thus, FG-4592 prevents α-syn-induced neurotoxicity through the reduction of oxidative stress by induction of HO-1.

Copyright © The Japanese Pharmacological Society1-B-P-046

一般演題（ポスター）

PAI-1 induction is a critical event for the onset of lipopolysaccharide-induced
acute kidney injury.

〇田中 恒輝1、尾花 理徳1,2、坂井 響1、禿 宏保1、山本 彩葉1、田中 翔大1、岡田 欣晃1、藤尾 慈1,2
大阪大・院薬・臨床薬効、2大阪大・先導的学際研究機構生命医科学融合フロンティア研究部

1【Background】
A systemic inflammatory response caused by sepsis leads to widespread organ dysfunction, including acutekidney injury. However, the pathogenetic mechanisms of acute kidney injury（AKI） in sepsis remain fully
elucidated. Recently, it has been reported that plasminogen activator inhibitor-1 (PAI-1) which is induced byinterleukin (IL)-6, plays a central role in thrombogenesis in septic patients. The aim of this study is to elucidate
the involvement of PAI-1 in septic AKI using lipopolysaccharide (LPS)-induce AKI model.
【Methods/Results】
C57BL/6J mice were intraperitoneally treated with LPS. LPS administration elevated IL-6 expression in sera.
Quantitative PCR demonstrated that the mRNA expression of

was increased in kidneys 6 hours after LPS

treatment. Administration of IL-6 to LPS model mice further increased PAI-1 expression compared to LPS
alone, accompanied by renal impairment. Finally, to investigate whether PAI-1 is involved in kidney injury,
TM5441, a PAI-1 inhibitor, was used. Administration of TM5441 suppressed urinary albumin/creatinine ratio, akidney injury marker, andmRNA expression, a tubular injury marker.

【Conclusion】
PAI-1 induction, which is potentiated by IL-6, contributes to the pathogenesis of LPS-induced AKI.

Copyright © The Japanese Pharmacological Society1-B-P-047

一般演題（ポスター）

Activation of FSGS-associated N-terminus-mutant TRPC6 channels bymechanical and receptor stimulations shows abnormal filtration barrier functionof mouse podocytes〇市川 純1、中川 緑2、井上 隆司2
1

佐野日本大学短期大学・総合キャリア教育学科・栄養士フィールド、2福岡大・医・生理学教室

Neurohormonal and mechanical responses exquisitely regulate various renal glomerular functions. Previously,
we found that mechanical stimulation reduces the activity of receptor -activated canonical transient receptor
potential 6 (TRPC6) channel in podocytes to enhance the glomerular barrier function. To explore its
pathological implications, we carried out the intracellular Ca2+ imaging, whole-cell patch clamp and albuminpermeation assay using immortalized mouse podocytes stably expressing wild-type (wt) TRPC6 or its Nterminus mutants associated with focal segmental glomerulosclerosis (FSGS) (P111Q, M131T and N142S).
Application of a membrane-expanding agent 2,4,6-trinitrophenol (TNP) immediately suppressed wt Ca2+
responses evoked by angiotensin II (Ang II). However, these responses were reversed in P111Q and entirely
absent in M131T and N142S. Simultaneous stimulation with Ang II and TNP in wt reduced FITC-labelled
albumin leak, while this was weakened in FSGS mutants. Pretreatment with a TRPC6-specific inhibitor
SAR7334 enhanced the leaks, the extent being greater in the mutants than wt. These results strongly suggestthat FSGS-associated N-terminus TRPC6 mutations may impair the filtration barrier function through thealtered efficiency of mechanical stress in suppressing receptor-activated TRPC6 channel activities.

Copyright © The Japanese Pharmacological Society1-B-P-048

一般演題（ポスター）

Effect of hyperphosphatemia on impairment of vascular function in adenineinduced renal injury in rats

〇小渕 修平、上田 紗夢、秋山 直樹、西畑 佑哉、上田 晴康
兵庫医科大・薬・薬理学

We reported that vascular function in adenine-induced renal injury rats was impaired mediated by increase inplasma indoxyl sulfate concentration. It is known that hyperphosphatemia leads to calcification andatherosclerosis. In the present study, we examined effect of hyperphosphatemia on vascular dysfunction in
adenine-induced renal injury in anesthetized rats. Renal injury was induced by feeding 0.75% adenine diet for 4
weeks. Lanthanum carbonate was treated with gavage after 2 weeks induced adenine. Treatment withlanthanum carbonate significantly decreased in plasma phosphorus concentration in adenine rats. N-nitro-Larginine (L-NA) potentiated the ACh-induced depressor response in normal-diet. However, L-NA failed topotentiate the response in adenine rats or lanthanum-treated rats. Sodium nitroprusside (SNP)-induced
depressor response in adenine rats was significantly smaller than that in normal rats. Lanthanum carbonate
tended to recover impairment of SNP-induced depressor responses in adenine rats. These findings suggest that
hyperphosphatemia is partially related to impairment of smooth muscle function rather than that of endothelial
function in adenine rats.

Copyright © The Japanese Pharmacological Society1-B-P-049

一般演題（ポスター）

Pharmacological inhibition of protein arginine methyltransferase 5 suppresses
TGF-β-induced fibrotic responses in cultured kidney fibroblasts〇茂木 飛佑馬1、刀坂 泰史1,2,3、砂川 陽一1,2,3、浜辺 俊秀1,2,3、小見山 麻紀2、長谷川 浩二1,2,3、森本 達也1,2,3
1

静県大・薬・分子病態、2京都医セ・展開医療、3静県総病院・臨床研究部

Although renal fibrosis is observed in chronic kidney disease, there is still no effective treatment
for it. Our previous study has shown that protein arginine methyltransferase 5 (PRMT5) is essential fortransforming growth factor-β (TGF-β)-induced transcription of fibrotic genes in cardiac fibroblasts. In thisstudy, we aimed to investigate the function of PRMT5 in renal fibrosis.
NRK-49f kidney fibroblast cells were stimulated with TGF-β for 48 hours. PRMT5 expression levels
were examined by qPCR and Western blotting (WB). Next, NRK-49f cells were transfected with siRNA of
PRMT5 followed by TGF-β stimulation and mRNA levels of myofibroblast marker α-smooth muscle actin (αSMA) and fibrotic genes (Col3a1, CTGF) were investigated by qPCR. Finally, NRK-49f cells were treated with
EPZ015666, a selective inhibitor of PRMT5 for 2 hours and then stimulated with TGF-β. The expression levelsof α-SMA were examined by qPCR and WB.
qPCR and WB revealed that the expression levels of PRMT5 were upregulated by TGF-β stimulation.
The expressions of α-SMA and the fibrosis-related genes were upregulated by TGF-β stimulation, whereas
knockdown of PRMT5 suppressed the upregulation of these genes. TGF-β-induced increase in α-SMA was
suppressed by treatment with EPZ015666.
Knockdown and pharmacological inhibition of PRMT5 suppressed TGF-β-induced myofibroblast
differentiation in kidney fibroblasts. These results suggest that PRMT5 is an essential molecule for TGF-βinduced fibrotic response in kidney fibroblasts.

Copyright © The Japanese Pharmacological Society1-B-P-050

一般演題（ポスター）

Protective effects of losartan on bladder dysfunction in spontaneously
hypertensive rats

〇清水 翔吾1、長尾 佳樹2、倉林 睦3、清水 孝洋1、東 洋一郎1、Zou Suo1、齊藤 源顕1
1

高知大・医・薬理、2高知大・医・小児思春期、3高知大・医・病理

Our previous report showed aging caused bladder dysfunction in spontaneously hypertensive rats
(SHRs). In this study, we investigated the protective effects of an antihypertensive drug, angiotensin II type 1
receptor blocker losartan on bladder dysfunction in aged SHRs.
Thirty-six-week-old male SHRs were orally treated with losartan (0, 3 or 10 mg/kg)
once daily for 18 weeks. Vehicle-treated Wistar Kyoto rats (WKYs) were used as normotensive controls.
Vehicle-treated SHRs had significantly higher detrusor thickness, bladder arterial wall thickness
compared to vehicle-treated WKYs. Moreover, SHRs showed significantly higher single voided volume (SVV),
post-voiding residual urine volume (RV), bladder capacity (BC), and intercontraction interval (ICI) and lower
voiding efficiency compared to vehicle-treated WKYs. A low dose of losartan decreased RV, BC and ICI but notmean blood pressure in SHRs. A high dose of losartan ameliorated changes in mean blood pressure, detrusor
thickness, bladder arterial wall thickness, SVV, RV, BC and ICI in SHRs.
Treatment with losartan ameliorated bladder dysfunction in aged SHRs.

Copyright © The Japanese Pharmacological Society1-B-P-051

一般演題（ポスター）

Inhibition of histone demethylation augments NAD synthesis through the PreissHandler pathway in cultured human proximal tubular epithelial cell line

〇ハサン アリフ1,2、丸茂 丈史2、小原 真美1、佐藤 幸子1、近藤 ゆき子1、村瀬 真⼀2、平 英一1
岩手医科大・医・薬理学講座情報伝達医学分野、2国際医療福祉大・医・薬理学

1

Boosting nicotinamide adenine dinucleotide (NAD) production shows beneficial effects against diabetic kidneydisease (DKD). However, the molecules that possess the therapeutic potential for increasing intracellular NADpool are not available. On the other hand, epigenetic modifiers are gradually gaining much interest in treating
diseases such as DKD. KDM1A demethylates Lys 4 and Lys 9 of histone H3. We hypothesize that a selective
KDM1A inhibitor, ORY-1001 may augment intracellular NAD synthesis in the DKD model. We induced
hyperglycemic milieu in human proximal tubular epithelial HK2 cell-line using high glucose (4.5 g/L) in thepresence or absence of ORY-1001 and compared the effect with their low glucose (1.0 g/L) containing
counterparts. As anticipated, high glucose significantly reduced cellular NAD, while increasing NADH contents.
The condition was restored by ORY-1001. To explore the underlying mechanisms, we thoroughly analyzed themRNA expression profiles of the genes that are involved in NAD synthesis and metabolism. Consistent with the
NAD production, NAPRT1 expression of the Preiss-Handler pathway was reduced by the high glucose, andtreatment with ORY-1001 restored the expression. Moreover, a similar expression profile was found for
PPARGC1a expression, a downstream effector of intracellular-NAD. Based on these findings, we presume that
the KDM1A-inhibitor ORY-1001 has potential therapeutic benefits against DKD through augmenting NAD
synthesis through upregulation of NAPRT1 of the Preiss-Handler pathway of NAD synthesis.

Copyright © The Japanese Pharmacological Society1-B-P-052

一般演題（ポスター）

Development of a SARS-CoV-2 infection model using human iPSC-derived
intestinal epithelium〇山田 茂1、野田 隆政2,3,4,5、岡部 かおり2、柳田 翔太1、西田 基宏6,7、諫田 泰成11

国立医薬品食品衛研、2国立精神・神経研、3国立精神・神経研、4国立精神・神経研、5東京慈恵会医科大、6九州大、7

生理学研

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has rapidly widespread and led to global health crises. COVID-19 causes well-known respiratory failure andgastrointestinal symptoms, such as diarrhea, nausea, and vomiting. Thus, human gastrointestinal cell model isurgently needed for COVID-19 research, nevertheless, primary human cells can be difficult to obtain. In thepresent study, we used small intestinal epithelial like cells (SIECs) from human induced pluripotent stem cells(iPSCs) for SARS-CoV-2 infection and drug testing. We observed that iPSC-SIECs, such as absorptive cells andpaneth cells, were infected with SARS-CoV-2. SARS-CoV-2 infection decreased transepithelial electricalresistance (TEER), an indicator of epithelial integrity. In addition, SARS-CoV-2 infection increased expression
levels of proinflammatory genes, which are elevated in patients with COVID-19. We further observed that
remdesivir treatment suppressed SARS-CoV-2 infection to SIECs. These data suggest that human iPSC-derived
SIECs provide a usefulmodel for elucidation of COVID-19 pathology.

Copyright © The Japanese Pharmacological Society1-B-P-053

一般演題（ポスター）

Allyl isothiocyanate-induced acute inflammation in the gastric mucosa leads tothe impaired gastric motility in rodents: involvement of substance P andcalcitonin gene-related peptide, but not mast cells〇田嶋 公人、大重 茉里奈、藤井 瑶子、堀江 俊治
城西国際大・薬We investigated the mechanism of acute inflammation induced by allyl isothiocyanate (AITC), a TRPA1
activator, through measurement of vascular permeability and gastric mucosal blood flow (GMBF) in rat
stomachs to provide the rodent model of impaired gastric motility. Male SD rats were used after 18 h-fasting.
chambers in anesthetized rats. Vascular permeability (extravasated

Stomachs were mounted on the

Evans blue) and GMBF (a leaser doppler) were measured in response to mucosal application of AITC. AITC
obviously increased vascular permeability and GMBF. Gastric mucosal swelling was observed after application of
AITC without hemorrhagic lesions. The both vascular permeability and GMBF in response to AITC weresignificantly attenuated by the pretreatment of a substance P receptor antagonist aprepitant, whereas GMBF
was alone significantly decreased by the pretreatment of a TRPA1 blocker A-967079 and a calcitonin generelated peptide (CGRP) receptor antagonist BIBN 4096. However, vascular permeability and GMBF in
response to AITC hardly affected by the pretreatment of a mast cell stabilizer cromoglycate. These results
suggest that AITC-induced acute inflammation with no mucosal damage is dependent on substance P and
CGRP released from TRPA1-expressing nerves in rat stomachs, but not mast cells. Those inflammation could
lead to the impaired motility in rodents.

Copyright © The Japanese Pharmacological Society1-B-P-054

一般演題（ポスター）

A dextran sodium sulfate-induced chronic colitis murine model of intestinal
fibrosis〇佐々木 礼一郎1、池田 一生2、天ヶ瀬 紀久子1,2
立命館大・院薬・病態薬理学研究室、2立命館大・薬・病態薬理学研究室

1

Chronic inflammatory disease such as Ulcerative Colitis (UC) and Crohnʻs Disease (CD) causes excessive
fibrosis and strictures in the intestine. An intestinal fibrosis model using dextran sodium sulfate (DSS)-induced
UC in mice was investigated to determine new treatment methods for UC. DSS containing drinking water was
given to C57BL/6N mice for 7 days, followed by 14 days of water (repeated three times). Animal body weight,
blood in the feces, fecal consistency, and disease activity index (DAI) were evaluated. To evaluate inflammation,
myeloperoxidase (MPO) activity was determined and hematoxylin and eosin (H&E) staining was performed.
Masson's trichrome, Picrosirius red staining, immunohistochemistry (IHC) and western blotting were used toevaluate the degree of fibrosis. The DSS group had higher DAI scores, a reduction in goblet cells, infiltration oflymphocytes, and higher MPO activity than the control group. Increased collagen fiber deposition in the colonand increased type I collagen in the lamina propria and submucosal tissue was evident in the DSS group, withmyofibroblasts in the lamina propria. Heat shock protein-47 and tissue inhibitor of metalloproteinases-1 weredetected. Repeated DSS intake exacerbates colitis with lymphocyte infiltration and fibrosis, with an increase intype I collagen in the lamina propria and submucosal tissue.

Copyright © The Japanese Pharmacological Society1-B-P-055

一般演題（ポスター）

Thromboxane A2 receptor signaling in macrophages attenuates acetaminopheninduced liver injury〇田邉 美奈1、伊藤 義也1、長田 真由子1、山下 敦1、古江 明子1、細野 加奈子1、畑中 公1、馬嶋 正隆2、天野 英樹11

北里大・院医療・分子薬理学、2神奈川工科大・健康医療

Over dosage of acetaminophen (APAP) administration causes severe acute liver failure.
Accumulating evidence suggests that macrophages contribute to APAP-induced liver injury; however,underlying mechanisms of involvement of macrophages remain unknown. We recently reported thatthromboxane A2 (TXA2) improves chemical-induced liver injury by accumulating macrophages. Here, we
examined the role of TXA2 in macrophages during APAP-induced liver injury.
APAP (300 mg/kg, ip) was administered to macrophage-specific thromboxane prostanoid
receptor (TP) deficient mice (mTPKO) and control mice (Cont). Compared with Cont, mTPKO exhibitedsevere liver injury as indicated by increased levels of ALT and necrotic area and decreased expression of PCNA,a marker of hepatocyte proliferation at 48 h post-APAP treatment. There was no statistical difference in hepatic
GSH levels between the two genotypes. TP and TXA2 synthase were expressed in CD68-positive cells in theliver. Immunofluorescence revealed CD68-positive cells accumulated extensively into the necrotic regions oflivers from Cont as compared with mTPKO. The expression of mRNA encoding pro-inflammatory mediators
including TNF-α, IL-1β and IL-6 in mTPKO were higher than in Cont, whereas HGF levels in mTPKO werelower than in Cont.
TP receptor signaling in macrophages attenuated APAP-induced liver injury by reducing
inflammatory cytokines and promoted liver repair by increasing macrophages in the necrotic regions and HGF
production.

Copyright © The Japanese Pharmacological Society1-B-P-056

一般演題（ポスター）

Orphan G protein-coupled receptor GPR35 contributes to the pathogenesis ofdextran sulfate sodium-induced colitis in mice.

〇岸 采花1、橘 佑輔1、村瀬 由依1、徳山 珖雅1、斉藤 美知子2、安田 浩之1、松本 健二郎1、加藤 伸一1
京都薬科大・病態薬科・薬物治療、2京都薬科大・バイオサイエンス研究セ

1Orphan G protein-coupled receptor GPR35, which is activated by lysophosphatidic acid and kynurenic acid, ishighly expressed in gastrointestinal tracts. Clinical findings indicate that this receptor has been implicated in theonset of inflammatory bowel disease (IBD), but its role in physiological and pathological processes in the colon
remains undefined. The present study investigated the role of GPR35 in the pathogenesis of exmerimentallyinduced colitis in mice.GPR35-deficient (GPR35KO) mice were generated by CRISPR-Cas9-mediated genome
editing on C57BL/6 background. Experimental colitis was induced in GPR35KO and wild-type (WT) miceby the treatment with dextran sulfate sodium (DSS) for 7 days. Lodoxamide, which has a GPR35
activating effect, was administered i.p. once daily for 7 days. DSS treatment produced body weight loss
with diarrhea and blood feces, and severe colitis characterized by shortening colon length and histological injury
7 days later. The severity of colitis with systemic symptoms was significantly augmented in GPR35KO micecompared with WT mice. In contrast, daily administration of lodoxamide significantly reduced the severity of
DSS-induced colitis in WT mice. However, the protective effect of lodoxamide was not observed in GPR35KO
mice. These findings suggest that GPR35 plays an anti-inflammatory role in DSS-induced colitis. Thus, GPR35
may be a promising target for treatment and prevention of inflammatory bowel disease.

Copyright © The Japanese Pharmacological Society1-B-P-057

一般演題（ポスター）
2+

Protective role of Ca -permeable TRPV6 in dextran sulfate sodium-induced
colitis in mice

〇森 風帆1、高山 麻由1、斉藤 美知子2、安田 浩之1、松本 健二郎1、加藤 伸一1
1

京都薬科大・病態薬科・薬物治療、2京都薬大・バイオサイエンス研究セTransient receptor potential vanilloid 6 (TRPV6), which is a highly Ca2+-selective ion channel, is expressed in
gastrointestinal epithelium and implicated in maintaining Ca2+ homeostasis via transcellular Ca2+ transport.
However, the local pathophysiological roles of TRPV6 in the gastrointestinal tract remains undefined. In thecurrent study, we investigated the role of TRPV6 in the pathogenesis of experimentally-induced colitis in mice.
TRPV6-deficient (TRPV6KO) mice were generated by CRISPR-Cas9-mediated genome editing on C57BL/6
background. Experimental colitis was induced in TRPV6KO and wild-type (WT) mice by the treatment withdextran sulfate sodium (DSS) for 7 days. Intestinal permeability was evaluated by FITC-dextran methods. DSS
treatment produced body weight loss with diarrhea and blood feces, and severe colitis characterized byshortening colon length and histological injury 7 days later. The severity of colitis with systemic symptoms was
significantly augmented in TRPV6KO mice compared with WT mice. Intestinal permeability was increased in
TRPV6KO compared with WT mice. The expression of E-cadherin and occludin in the colon was reduced in
TRPV6KO mice compared with WT mice. These findings suggest that TRPV6 plays a protective role in thepathogenesis of DSS-induced colitis via maintaining colonic barrier functions. Thus, TRPV6 may be a novel
target for the treatment of gastrointestinal diseases.

Copyright © The Japanese Pharmacological Society1-B-P-058

一般演題（ポスター）

Functional expression of calcium-sensitive receptors in activated hepatic stellate
cells〇近藤 るびい、川田 成紀、鈴木 良明、山村 寿男
名古屋市立大・院薬・細胞分子薬効解析学

Hepatic stellate cells (HSC) are liver-specific fibroblasts that play a critical role in the development of hepatic
fibrosis. During liver injury, these cells transdifferentiate into the activated phenotype, resulting in enhanced cellproliferation and extracellular matrix production. The functions of activated HSCs require an increase incytosolic Ca2+ concentration ([Ca2+]cyt). However, the regulatory mechanisms underlying Ca2+ signaling in
activated HSCs remain largely unknown. In the present study, the pathophysiological roles of calcium sensing
receptors (CaSRs) were examined in human hepatic stellate cells LX-2. Expression analyses revealed that CaSR
proteins were expressed in α-smooth muscle actin-positive LX-2 cells. Extracellular Ca2+ restoration (from 0 to
2.2 mM) increased [Ca2+]cyt in LX-2 cells. The extracellular Ca2+-induced increase in [Ca2+]cyt was reduced by the
CaSR antagonists, 10µM NPS2143 and Calhex 231.TGF-β1 caused the upregulation of myofibroblast markers,
α-SMA and Col1α1, in LX-2 cells. This upregulation was markedly reduced by NS2143. The treatment with
NS2143 and Calhex 231 significantly attenuated the proliferation of LX-2 cells. These results indicate that
CaSRs are functionally expressed in hepatic stellate cells and contribute to extracellular matrix production andcell proliferation.

Copyright © The Japanese Pharmacological Society1-B-P-059

一般演題（ポスター）

The role of Id4 in salivary gland and its involvement in the pathology of IgG4related disease〇木村 宗惟1,2、林 慶和1,3、矢野 恵奈1、佐伯 彩華1、安河内 篤2、森山 雅文2、中村 誠司2、自見 英治郎1、安河内（川久保） 友世1
1

九州大・院歯・OBT研究セ、2九州大・院歯・口腔顎顔面病態学・顎顔面腫瘍制御学、3福岡歯科大・歯・生体構造学・機能構造

Id (inhibitor of DNA binding/differentiation), a group of dominant negative transcriptional regulators for basic
helix-loop-helix transcription factors, consists of Id1-Id4. Previous studies showed Id proteins are involved incell differentiation and proliferation, and those deficiency leads various pathological conditions. However, thephysiological functions of Id4 have not been clarified. We thus investigated the role of Id4 in the salivary gland.
In this study, we first analyzed the impact of Id4 on salivary glands using Id4 deficient (submandibular glands (SMG) weight and saliva secretion inKO) mice. The

KO mice were significantly decreased comparedto those in wild-type mice. Histological analysis revealed that increased expressions of various differentiation
markers and significant mucus accumulation were observed in the SMG of

KO mice.

Subsequently, we investigated the possibility that Id4 is involved in human pathology, such as Sjögren
syndromeand IgG4-RD in which saliva secretion often decreases. As a result, the expression level of Id4 wassignificantly decreased in the SMG tissue of IgG4-RD, and miRNA-mRNA integrated analysis using humansamples revealed that Id4 might be downregulated by hsa-miR-486-5p in IgG4-RD salivary glands.
Taken together, we suggest that Id4 is essential for the homeostasis of salivary glands, and miR-486-5p, as wellas Id4, might be associated with the pathophysiology of IgG4-RD.

Copyright © The Japanese Pharmacological Society1-B-P-060

一般演題（ポスター）

Pharmacological analysis of salivary secretion mechanisms using rats withdifferent strains of Aquaporin 5 levels.

〇根津 顕弘、Akter MST Tahmina、谷村 明彦
北海道医療大・歯Aquaporin 5 (AQP5) plays an important role for the transcellular fluid secretion in salivary gland cells. In thisstudy, we compared acetylcholine (ACh)-induced salivary secretion and blood flow (BF) dynamics insubmandibular gland (SMG) in rat strain with low levels of AQP5 protein (AQP5/low), Sprague-Dawley (SD)and Wistar/ST rats.The whole saliva secretion with high-dose of ACh (720~1440 nmol/min) in AQP5/low andWistar/ST were~70% of that in SD. The level of AQP5 protein in Wistar/ST was same as that in AQP5/low,
and was much lower than that in SD, suggesting that the AQP5 level determine the maximum rate of salivary
secretions.Interestingly, the salivary secretions with low-dose of ACh (60~120 nmol/min) in Wistar/ST was twotimes higher than that of AQP5/low, and was comparable to that in SD. The ED50 values for ACh-induced
salivary secretion in AQP5/low, Wistar/ST, and SD were 309, 102, and 134 nmol/min, respectively. These
results suggest that ACh sensitivity in salivary secretion does not correlate with AQP5 levels. Monitoring of BF
in SMG demonstrated that low-dose of ACh induced oscillatory changes in BF in all strains. The BF oscillations
in Wistar/ST wereobservedmostlyabove the resting level, whereas that in AQP5/low were observed below theresting level. We also found that angiotensin II-mediated vasoconstriction reduced BF during ACh stimulations,
resulting in a decrease insalivary secretionin AQP5/low.These results suggest that the regulation of salivary
secretionwith physiological stimuli can be determined by BF rather than by the level of AQP5.

Copyright © The Japanese Pharmacological Society1-B-P-061

一般演題（ポスター）

In vivo monitoring of increase in norepinephrine and serotonin in the interstitial
fluids in rat submandibular glands by perfusion with imipramine〇白勢 康介1、姜 卓義1、渡邊 真理子1、松田 光正1、伊藤 健二1、鈴木 武志1、小林 広幸2、吉川 正信2
東海大・医・麻酔科学、2東海大・医・臨床薬理学

1

Previous studies demonstrated contents of monoamines in homogenate obtained from salivary glands.
Monoamines in homogenate, however, includes that stored in the cells, as well as that released in the interstitial
fluids. So far, direct monitoring of autonomic nervous activity within salivary glands has been difficult, and therelationship between neurotransmitter release in the salivary gland and salivary secretion has not been analyzed.
Microdialysis was applied to salivary glands and successfully monitored multiple neurotransmitterssimultaneously. The present study examined the effects of imipramine, a tricyclic antidepressant, on contents of
monoamines in interstitial fluids within submandibular glands of rats. The results revealed the following: (1)that microdialysis allowed detection of norepinephrine and serotonin, but not epinephrine or dopamine; (2) that
their concentrations in the dialysate were highly variable and unstable over the first 120 min after probe
implantation, but reached a nearly stable level thereafter; and (3) that perfusion with imipramine significantly
and dose-dependently increased norepinephrine and serotonin concentrations in the dialysate. These results
indicate that the present microdialysis technique offers a powerful tool for detecting changes in sympathetic
activity within the salivary glands.

Copyright © The Japanese Pharmacological Society1-B-P-062

一般演題（ポスター）

Arginase 1 might regulate the exocrine secretion via metabolism〇大野 雄太1、長瀬 春奈1、佐藤 慶太郎2、設楽 彰子1、中本 哲自3、柏俣 正典11

朝日大・歯・歯科薬理、2明海大・歯・薬理、3朝日大・歯・インプラント

Sjogrenʻs syndrome induces salivary and lacrimal hyposecretion, which leads to reduced quality of life. Althoughmany studies have been conducted from the perspective of inflammation, the development of a causal treatment
has not been achieved yet. We previously investigated the cause from the perspective of non-inflammation, andidentified arginase 1 as a novel non-inflammatory regulator of exocrine function. However, the mechanism ofarginase 1 regulating the function remains unknown, so we aimed to elucidate the mechanism.
We first confirmed the expression of arginase 1 in both salivary and lacrimal glands from BALB/c mice. Arginase
1 inhibitor, CB-1158, reduced the pilocarpine-induced saliva and tear secretion. The metabolome

analysis of lacrimal glands revealed the altered concentration of amino acids related to energy metabolism.
Furthermore, in an

perfusion system in which submandibular glands removed from the CB-1158-treatedmice were cannulated with arteries, the saliva flow rate immediately after stimulation was unchanged comparedto the control group but then decreased. We did not observe the saliva decrease inexperiments withperfusion of CB-1158 itself. These results indicated that arginase 1 might regulate paracellular fluid secretion
energy metabolism.

Copyright © The Japanese Pharmacological Society1-B-P-063

一般演題（ポスター）

Examination of making a cylindrical defect model of rabbit meniscus〇岸浪 昌礼、鈴木 陽子、佐々木 麻衣、西堀 頼史、山崎 則之㈱新薬リサーチセンター・非臨床研究部【Background and Objective】
We have created a rabbit osteoarthritis model by partial resection of the lateral meniscus of the left hind limb
knee joint. This time, we created a cylindrical defect model of the meniscus by biopsy trepan using a Japanesewhite rabbit, and compared the condition of the meniscus and the condyle joint 28 days after the operation.
【Method】
A hole was made in the center of the medial meniscus of the right knee joint of 4 Japanese white rabbits
weighing 2.9 to 3.4 kg using biopsy trepan (φ1.0 and 2.0 mm). The tibial condyle and meniscus were removed
and visually observed.
【Result】
In the φ1 mm cylindrical defect model, the meniscus remained punctured.
The φ2 mm cylindrical defect model also had holes in the meniscus.
We compared the damage to the cartilage of the femoral condyle and tibial condyle with the partial meniscal
resection model.
【Conclusion】
This model was also considered to be effective as a meniscus repair test.

Copyright © The Japanese Pharmacological Society1-B-P-064

一般演題（ポスター）

Lactoferrin improves the reduction of bone formation in Dexamethasone-induced
osteopenia mice

〇青木 亮憲1、古川 恵1、東方 優大2、伊藤 芳久1、大野 恵3、出雲 信夫2,41

横浜薬科大・薬・薬学教育セ、2横浜薬科大・薬・薬物治療学研、3（株）NRLファーマ、4横浜薬科大・薬・総合健康メディ

カル研究セ【Objective.】
Steroids are used in clinical practice for a variety of conditions, including immunosuppression. However,steroid-induced osteoporosis due to decreased bone formation (De Nijs., 2008) is a problem (Suzuki et al.,2014). Lactoferrin (LF) is a protein found in breast milk and other sources and has been reported to promoteosteoblast differentiation (Icriverzi et al., 2020). In this study, we investigated the therapeutic effect of LF on
dexamethasone (DEX)-induced bone loss.
【method】
8-week-old ddy male mice were treated with DEX (2 mg/kg) for 8 weeks. 4 weeks after DEX administration, LF
was administered by forced oral administration at a dose of 100 or 300 mg/kg for 6 days per week. 4 weeks after
LF administration, femur and tibia were removed and bone strength measurements, CT measurements andmRNA The gene expression levels of bone metabolism markers were examined using RT-PCR.
【Results and Conclusions】
Bone strength after 8 weeks of DEX treatment was significantly lower than that of the untreated group, and LF
administration ameliorated the decrease in bone strength. Similarly, CT results showed that LF administration
significantly improved the DEX-induced decrease in bone mineral content and bone mineral density. RT-PCR
results showed that the gene expression levels of osteogenic markers were significantly decreased by DEX
administration and significantly improved by LF administration. These results suggest that LF improves theinhibition of bone loss by DEX through the involvement of the osteogenic system.

Copyright © The Japanese Pharmacological Society1-B-P-065

一般演題（ポスター）

Cdc42〇長瀬 春奈1、大野 雄太1、佐藤 慶太郎2、柏俣 正典1、設楽 彰子1
朝日大・歯・薬理、2明海大・歯・薬理1Epithelial cells of exocrine glands responsible for saliva and tear secretion bear cell polarity. Cdc42, essential forthe polarity of epithelial cells, is required for the formation and maintenance of luminal structures which isimportant site for the secretion. However, it is still unclear whether Cdc42 plays the identical role in different
epithelial tissues. In this study, we generated exocrine epithelial cell-specificconditional knockout(KO) mice and analyzed the difference of Cdc42 roles between salivary glands and lacrimal glands.
Morphological analysis showed that luminal structures changed to thick, short, bulging structures in the both
KO glands. Since these glands weight decreased, we analyzed inflammation with HE staining andapoptotic cell death with TUNEL staining. Both glands showed no inflammation nevertheless TUNEL (+) cells
increased, suggesting disrupted cell polarity. In contrast, pilocarpine-stimulated saliva secretion decreased whiletear secretion increased. Moreover, protein expression of AQP5, essential for exocrine fluid, also decreased in
the salivary glands but conversely increased in lacrimal glands.
These findings suggest that Cdc42 in salivary glands and lacrimal glands plays an identical role in epithelial cell
polarity formation but an opposite role in the secretory mechanism.

Copyright © The Japanese Pharmacological Society1-B-P-066

一般演題（ポスター）

mPGES-1 promotes granulation tissue angiogenesis through regulatory T-cellaccumulation〇兵頭 徹也1,2、天野 英樹1、伊藤 義也1、細野 加奈子1、畑中 公1、江島 耕二3、林 泉4、植松 智5、審良 静男6、武田 啓
2、馬嶋 正隆71

北里大・医・薬理学、2北里大・医・形美学、3北里大・医・免疫学、4日本薬科大・医療薬学科、5大阪市立大・院医・医・ゲ

ノム免疫学、6大阪大・免・フロンティア研究センター、7神工大・健医学・臨工科病態治療研究室

Microsomal prostaglandin E synthase-1 (mPGES-1) is an enzyme responsible for the final step of prostaglandin
E2 (PGE2) synthesis. PGE2 involves in wound-induced angiogenesis. Regulatory T cells (Tregs) regulate not
only immune tolerance but also tissue repair and angiogenesis. Herein, we examined whether the mPGES-1/PGE2 axis contributes to wound-induced angiogenesis and granulation tissue formation through Treg
accumulation. Polyurethane sponge disks were implanted into the dorsal subcutaneous tissues of the male
mPGES-1-deficient (mPGES-1-/-) and C57BL/6 wild-type (WT) mice. Compared with WT mice, angiogenesis
was suppressed in mPGES-1-/- mice, which was associated with attenuated forkhead box P3 (Foxp3) expression
and Foxp3+ Treg accumulation. The numbers of double-positive cells for Foxp3/TGFβ and Foxp3/VEGF werelower in mPGES-1-/- mice than in WT mice. Deleting Tregs with neutralizing antibodies (Abs) against CD25
or folate receptor 4 (FR4) inhibited the Foxp3+ Treg angiogenesis and accumulation in WT mice but not in
mPGES-1-/- mice. The topical application of PGE2 into the implanted sponge enhanced Treg angiogenesis andaccumulation expressing TGFβ and VEGF in WT and mPGES-1-/- mice. These results suggest that mPGES-1-derived PGE2 promotes wound-induced angiogenesis, at least in part, by producing TGFβ and VEGF in
accumulated Tregs. mPGES-1 induction would control angiogenesis with Treg recruitment in skin wounds.

Copyright © The Japanese Pharmacological Society1-B-P-067

一般演題（ポスター）

P2X4 receptor signal enhances mast cell activation by Mas-relate G proteincoupled receptor b2 in a phosphatidylinositol-3-kinase-dependent manner〇吉田 一貴、大林 晃右、伊藤 政明、松岡 功
高崎健康福祉大・薬

Mas-relate G protein-coupled receptor X2 (MRGPRX2) and its mouse ortholog Mrgprb2 are specifically
expressed in mast cells and involved in pseudoallergic reactions. We previously reported that extracellular ATP
augmented Mrgprb2-mediated mast cell activation via P2X4 receptors. In this study, we investigated themechanism underlying the synergistic effects of co-stimulation of Mrgprb2 and P2X4 receptor on degranulationin mouse peritoneal mast cells (PMCs). Stimulation of Mrgprb2 with compound 48/80 induced degranulationaccompanied by an increase in intracellular Ca2+ concentration ([Ca2+]i). ATP also induced a rapid increase in[Ca2+]i. Co-stimulation with ATP and compound 48/80 promoted the sustained increase in [Ca2+]i. Thissynergistic Ca2+ response was absent in PMCs prepared from P2X4 receptor deficient mice. In addition, bothincreased degranulation and synergistic Ca2+ response to co-stimulation with ATP and compound 48/80 were
inhibited by the PI3K inhibitors wortmannin and AS605240. These results suggested that the P2X4 receptor
signal promotes Mrgprb2-induced degranulation and Ca2+ response in a PI3K-dependent mechanism.

Copyright © The Japanese Pharmacological Society1-B-P-068

一般演題（ポスター）

CDK8/19 inhibitors induce M2-like macrophage polarization.

〇水野 夏実、志賀 咲紀、柳川 芳毅
北海道医療大・薬

Macrophages polarize into anti-inflammatory macrophages (M2 macrophages) by interleukin (IL)-4, and theseM2 macrophages express arginase-1. In addition, it has been reported that several cyclin-dependent kinase(CDK) 8/19 inhibitors promotes anti-inflammatory responses, and a number of CDK8/19 inhibitors have beendeveloped. However, the effects of CDK8/19 inhibitors on arginase-1 expression in macrophages have not beenclarified. In this study, we investigated the effects of CDK8/19 inhibitors on arginase-1 expression in murine
macrophage cell line RAW264.7. The cells were pre-treated with BRD6989 or Senexin A, a CDK8/19 selective
inhibitor, and then stimulated with IL-4. BRD6989 and Senexin A increased the IL-4-induced arginase-1
expression. Furthermore, we founded that p38 MAPK inhibitors suppressed the BRD6989-increased arginase-1
expression. On the other hands, BRD6989 and Senexin A increased the surface expression of CD206, a M2
macrophage marker, in RAW264.7 cells. In conclusion, we demonstrated for the first time that CDK8/19
inhibitors increased arginase-1 expression in macrophages via p38 MAPK activation. These findings suggestthat CDK8/19 inhibition might induce anti-inflammatory M2-like macrophages.

Copyright © The Japanese Pharmacological Society1-B-P-069

一般演題（ポスター）

Analysis of mechanism underlying the synergistic inflammatory cytokine
production via P2X4 and EP3 receptors in mast cells

大林 昂右、吉田 一貴、朝比奈 愛理、内田 真耶子、伊藤 政明、〇松岡 功
高崎健康福祉大・薬

Mast cells (MCs) produce a variety of chemokines and cytokines to induce allergic inflammation. Previously, we
have shown that co-stimulation with ATP and prostaglandin (PG) E2 synergistically increases the secretion ofvarious inflammatory cytokines via P2X4 and EP3 receptors, respectively. In the present study, we examined themechanism underlying the synergistic cytokine production by co-stimulation of P2X4 and EP3 receptors inmouse bone marrow-derived MCs (BMMCs). Stimulation of BMMCs with PGE2 elicited rapid phosphorylationof ERK1/2, p38 MAP kinase, and Akt. In contrast, ATP alone had only weak effects on phosphorylation of thesesignaling molecules, and little affected PGE2-induced responses. Although ATP and PGE2 hardly induced NFkB p65 phosphorylation, co-stimulation with them elicited an increase in NF-kB p65 phosphorylation. Thiseffect was absent in BMMCs obtained from P2X4 receptor deficient mice. The cytokine production and NF-kB
p65 phosphorylation induced by co-stimulation with ATP and PGE2 were suppressed by inhibitors of NF-kB
and phosphatidylinositol-3-kinase (PI3K). These results suggest that co-stimulation of P2X4 and EP3 receptors
enhances MC cytokine production by activating NF-kB signaling pathway in a PI3K-dependent manner

Copyright © The Japanese Pharmacological Society1-B-P-070

一般演題（ポスター）

Expression and distribution of immune checkpoint molecules LAG-3 in
amyotrophic lateral sclerosis〇大島 基希1、青木 拓門1、青野 らん1、森﨑 祐太1、山中 宏二2、三澤 日出巳11

慶應義塾大・薬・薬理学講座、2名古屋大・環境医学研究所 病態神経科学分野

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by the selective
death of motor neurons. As a mechanism of motor neurodegeneration, neuroinflammation by immune cells suchas glial cells and other peripheral immune cells infiltrating in the central nervous system has been suggested.
However, it is still unclear how the immune cell activation is regulated. We focused on immune checkpoint
molecules (e.g. PD-1 and LAG-3) and analyzed their expression in ALS model mice (SOD1G93A mice). After the
disease onset, when compared to WT mice, SOD1G93A mice showed an increased expression of membrane andsoluble forms of LAG-3 in the spinal cord, but no change was detected in LAG-3 expression in peripheral tissues
such as spleen and lymph nodes. We performed immunohistochemical analyses in the spinal cord of SOD1G93A
mice and found that LAG-3 was expressed in microglia. Using the mouse microglial cell line BV2, we analyzed
changes in LAG-3 expression upon differentiation into inflammatory (M1)/anti-inflammatory (M2) state. We
found that the LAG-3 mRNA expression levels were upregulated in M1 microglia, but not in M2 microglia.
Elucidation of LAG-3 function in microglia will lead to a better understanding of the involvement of theimmune checkpoint molecule in the pathophysiology of ALS.

Copyright © The Japanese Pharmacological Society1-B-P-071

一般演題（ポスター）

Acute stress exacerbates itch-related scratching in atopic dermatitis mice:possible involvement of allopregnanolone in the brain〇藤井 正徳、今林 怜矢、岸 美羽、山根 優、田中 里奈
京都薬科大Itch is the most bothersome symptom of atopic dermatitis and is often exacerbated by stress. Allopregnanolone(ALLO), one of the neurosteroids in the brain, is shown to increase rapidly following acute stress, serving as ahomeostatic response to stress. We have previously demonstrated that an intracisternal injection of ALLO
markedly increases itch-related scratching behavior in atopic dermatitis model mice. The present study
examines whether acute stress exacerbates itching in those mice by comparing scratching behavior under threedifferent conditions: 1) conventional conditions (5 mice in the housing cage), 2) confinement stress (underisolated conditions in a small invisible chamber), and 3) confinement stress + forced swim stress (FSS) (underisolated conditions in the same chamber immediately following FSS). Scratching was significantly increased
under confinement stress conditions compared to conventional conditions. FSS further exacerbated scratching
behavior. Our results show that acute stress exacerbates itch-related scratching in atopic dermatitis mice. This
exacerbation of scratching may involve ALLO transiently increased in the brain as a response to stress.

Copyright © The Japanese Pharmacological Society1-B-P-072

一般演題（ポスター）

Involvement of neuropeptide; Gastrin-releasing peptide (GRP) and Galanin (GAL)
in allergic rhinitis model mice.

〇木村 徹1、横井 秀格2、松本 祐磨2、川田 往嗣2、齋藤 康一郎2、櫻井 裕之1
1

杏林大・医・薬理、2杏林大・医・耳鼻Allergic rhinitis (AR) is caused by an allergic reaction at nasal epithelia against allergens such as pollen andhouse dust. The antihistamine drugs or leukotriene receptor antagonist are used for the AR therapy, however
there are some patients, who do not respond to these drugs. Hence, it needs to develop newer drugs. In thisstudy, we focused on neuropeptides known to be involved in an inflammatory process: Gastrin-releasing peptide(GRP) and Galanin (GAL).
AR model mice were constructed as follows. The ovalbumin together with the adjuvant, aluminum hydroxide
was injected into mice intraperitoneally three times every other week as primary sensitization. Then ovalbumin
was administrated intraperitoneally 14 consecutive days as secondary sensitization. Allergic reactions were
evaluated by the number of rubbing sneezing episodes.
GRP and its receptor, GEPR, were expressed in nasal epithelial cells and the mast cells, and their expression wasincreased after AR induction. A GRPR antagonist suppressed AR symptoms. GAL and its receptor, GALR2
were expressed in nasal epithelial cells and B cells. A GALR2 antagonist suppressed the level of serum IgE,
numbers of B cells. It seems that the signaling of GRP and GAL may be involved in the pathophysiology of AR
and their inhibition would become the new therapeutic target for AR.

Copyright © The Japanese Pharmacological Society1-B-P-073

一般演題（ポスター）

Effects of extracellular nucleotides on macrophage activation induced bystretch-mediated mechanical stimulation〇伊藤 政明、長谷川 敦也、大熊 範和、吉田 一貴、松岡 功
高崎健康福祉大・薬

Although the immune cells experience mechanical forces and pressures throughout their life cycle, little is
known about how such mechanical processes regulate the immune cell function. We previously reported thatcyclical stretch (CS) stimulation of murine macrophage RAW264.7 (RAW) cells evoked release of nucleotides
including ATP and also triggered elevation of mRNAs for various pro-inflammatory factors. In this study, weinvestigated the role of extracellular nucleotides on RAW cells activation induced by CS stimulation.
In RAW cells, CS stimulation evoked a marked release of ATP and also elevated mRNA and protein levels formonocyte chemoattractant protein-1 (MCP-1). Direct stimulation of RAW cells with extracellular nucleotides
also triggered MCP-1 mRNA expression with a rank order efficacy: UTP, UDP >> ATP. RAW cells expressed
functional P2 receptors including P2Y6 receptor for uridine nucleotide. In the presence of P2Y6 receptor
antagonists, CS-induced MCP-1 expression was suppressed. In addition, P2Y6 receptor gene knock-down withsiRNA reduced CS-induced MCP-1 mRNA elevation without affecting ATP release.
These results suggest that P2Y6 receptor activation via autocrine stimulation by released extracellular
nucleotides may be involved in CS-induced MCP-1 expression in macrophages.

Copyright © The Japanese Pharmacological Society1-B-P-333

一般演題(ポスター)

Changes in the expression of interferon-induced transmembrane protein-3(IFITM3) in the brains of Alzheimer‘s disease model mice

〇LIU YUE1、溝口 博之1,2、祖父江 顕3、山中 宏二3、山田 清文1
名古屋大・院医・医療薬学、2名古屋大・環境医学研究所・MIRAIC-未来の医学研究センター、3名古屋大・環境医学研

1究所・病態神経科学

Interferon-induced transmembrane protein-3 (IFITM3) belongs to the IFITM family, which comprises five andseven subtypes in humans and mice, respectively. IFITM proteins participate in various biological processes
such as the immune response, including suppression of viral infection. While the immune response is associatedwith the pathology and development of Alzheimerʻs disease (AD), it is unknown whether this response isbeneficial or harmful. Recently, IFITM3 was found to be a γ-secretase modulatory protein, a type of protein
associated with the generation of amyloid b (Aβ). However, further research is needed to determine whether
IFITM3 is associated with abnormal behaviors, including cognitive impairment in AD, and whether it may be a
new molecular target for AD therapy. Since our final goal is to clarify the role of IFITM3 in an animal model ofAD, in this study we used

mice, which overproduce Aβ-42 without overexpressing amyloid precursor

protein, to examine changes in IFITM3 expression. The cortex, dentate gyrus (DG), and CA3 regions of 4- and8-month-old

mice exhibited Aβ accumulation and also increased IFITM3 expression. Expressions ofastrocytes and microglia were age-dependently increased around Aβ plaques. Notably, IFITM3 expression
colocalized with astrocytes, which were situated near Aβ in all brain regions of 8-month-old

mice.

These results suggest that IFITM3 is increased in astrocytes and is accompanied by Aβ accumulation.

Copyright © The Japanese Pharmacological Society1-B-P-074

一般演題（ポスター）

Cigarette smoke extract derived from heated tobacco products promotes cancer
stem cell properties of lung cancer cell lines.

〇平田 尚也1、堀之内 孝広2、諫田 泰成1
1

国衛研・薬理、2北海道大・院医・細胞薬理

Cigarette smoking is a risk factor for carcinogenesis and the development of several types of cancer, including
lung cancer. Cigarette smoke is considered to contain over 5,000 chemicals, including carcinogens. Heated
tobacco products (HTPs) have been recently reported to reduce levels of toxic chemicals, such as nicotine andtobacco-specific N-nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), compared to burned
cigarettes. We have previously reported that nicotine and NNK induce the proliferation of cancer stem cells(CSCs), which contribute to tumorigenicity, drug resistance and recurrence. To evaluated whether HTPs are
involved in cancer development, we examined the effects of cigarette smoke extract (CSE) derived from HTPs
on lung CSCs. We found that CSE induced the proliferation of lung CSCs and increased the expression levels ofstem cell markers. Since CSCs exhibit epithelial-mesenchymal transition (EMT), we analyzed the expression of
EMT markers. CSE induced expression of EMT markers, such as Twist and Snail. These results suggest that
HTPs can promote CSC properties in vitro and may result in cancer development.

Copyright © The Japanese Pharmacological Society1-B-P-075

一般演題（ポスター）

Doxorubicin did not affect the lysosomal acidification〇佐藤 岳哉1,3、戸田 法子1,2、斎藤 将樹3、山内 正憲2、阿部 高明1
1

東北大・院医・病態液性制御学分野、2東北大・院医・麻酔科学・周術期医学、3東北大・院医・分子薬理学分野

Doxorubicin (Dox) has been used as an effective antitumor agent against various types of cancers. However, its
application is limited owing to severe cardiomyopathy. Many researchers have attempted to clarify themechanism of DOX-induced cardiomyopathy; however, its underlying mechanisms remain unclear. We showedthat Dox impaired autophagic flux. In this study, we focused on the effect of Dox on lysosomes, a prerequisiteorganelle for autophagy that degrades autophagic cargoes. We speculated that Dox might inhibit autophagy byinhibiting the lysosomal function. To evaluate the effect of Dox on lysosomal acidification, we examined theeffect of Dox on lysosomal acidification by staining cells with the acidophilic fluorescent dye acridine orange todetermine the pH of lysosomes. Acridine orange emits red fluorescence under acidic conditions, whereas itemits green fluorescence under neutral pH conditions. The green and red fluorescence intensities of acridine
orange were measured by flow cytometry. Bafilomycin, an H+-pump inhibitor of lysosomes, significantly
inhibited lysosomal acidification. In contrast, lysosomal acidification increased considerably after Dox treatment.
We further explored the effect of Dox on the expression of transcription factor EB (TFEB), which is known toregulate lysosomal function. Unexpectedly, Dox did not affect the expression of TFEB. These results indicatedthat Dox does not affect lysosomal acidification or function.

Copyright © The Japanese Pharmacological Society1-B-P-076

一般演題（ポスター）

Phospholipase C like protein PRIP1 PH-domain-containing liposomes enhance
apoptotic cell death in cisplatin resistant breast cancer cells〇浅野 智志1、吾郷 由希夫1、兼松 隆1,2
広島大・院医系科学・細胞分子薬理、2九州大・院歯・口腔細胞工学

1Cisplatin is one of the most frequently used chemotherapeutic agents for the treatment of several humanmalignancies, and induces caspase-9-mediated apoptosis. Here, we examined whether phospholipase C-related
catalytically inactive protein (PRIP) enhances cisplatin-induced apoptosis of cisplatin resistant breast cancer
cells. PRIP depletion increased expression of X-linked inhibitor of apoptosis protein (XIAP) by inhibiting
protein degradation, which is downstream of PI3K/AKT pathway and inhibits apoptotic signaling by blocking
caspase-9 activation. Conversely, XIAP was decreased by expression of PRIP1 or pleckstrin homology domain of
PRIP1 (PRIP1-PH domain) that blocked PI(4,5)P2 metabolism. The expression levels of cleaved caspase-9 anddownstream cleaved caspase-7 and poly-ADP ribose polymerase were greater in PRIP1 or PRIP1-PH domainexpressing MCF-7 cells treated with cisplatin than in control cells. In an orthotopic transplantation model,
combined administration of PRIP1-PH domain-containing liposomes and cisplatin reduced the size of MCF-7
tumors compared with cisplatin alone. Our findings demonstrate that PRIP promotes XIAP degradation by
inhibiting PI(3,4,5)P3/AKT signaling and enhances cisplatin-induced apoptotic cell death. Therefore, we
propose that PRIP1-PH-liposomes are a novel agent to avoid cisplatin resistance.

Copyright © The Japanese Pharmacological Society1-B-P-077

一般演題（ポスター）

Differentiation inducing factor-1 suppressed the epithelial-mesenchymaltransition via degradation of Yes-associated protein〇高橋 富美、有岡 将基、岸上 赳大、石兼 真
産業医大・医・薬理Differentiation-inducing factors (DIFs), produced by, show anti-tumor activity via

several different signaling molecules, including glycogen synthase kinase-3 (GSK-3) and signal transducer andactivator of transcription 3 (STAT3). However, the exact mechanism of action of DIFs is still poorly understood.
Therefore, to better understand the action of DIF, we performed DNA microarray analysis and found that theactivity of the Hippo signaling pathway may be modified. Therefore, we analyzed mRNA expressions of Hippo
signaling pathway target genes. Although the mRNA expression of the target genes including CTGF and
Cyr61were elevated by DIF-1, surprisingly the protein expression themselves were reduced. In an attempt toelucidate the mechanism, we found that DIF promoted proteolysis of Yes-associated protein (YAP), atranscriptional co-activator of the Hippo signaling pathway, in the human cervical cell line HeLa and the humancolon cancer cell line HCT-116. YAP has also been reported to promote epithelial-to-mesenchymal transition(EMT). Indeed, cell migration, cell invasion and expressions of EMT-related proteins (fibronectin, vimentin andN-cadherin) were reduced by DIF-1. These results suggest that DIF-1 suppresses EMT via degradation of YAP
in human cancer cell lines.

Copyright © The Japanese Pharmacological Society1-B-P-078

一般演題（ポスター）

Probenecid has diverse effects on 3D-cultured prostate cancer cells.

〇宇和田 淳介、中澤 瞳、益岡 尚由
金沢医科大・医・薬理学

Probenecid, an uricosuric drug, has diverse pharmacological targets, including multidrug resistance-associatedprotein (MRP). Since inhibition of MRPs contributes to suppression of efflux of anti-cancer drugs, probenecid
has been investigated for cancer therapy as a chemosensitizer. In this study, we examined the effects ofprobenecid on 3D-cultured prostate cancer cells. In the 3D-cultured spheroids of 22Rv1 cells that were lesssensitive to cisplatin and doxorubicin than the 2D-cultured monolayer cells, probenecid treatment (100 and 300μM) increased sensitivity to those anti-cancer drugs. On the other hand, a higher concentration (500 μM) ofprobenecid showed no chemosensitizing effect, which are consistent with increase in ABCG2, a drug-efflux
transporter, at the dose. Furthermore, we found that probenecid has various anti-cancer effects other thanalteration of chemosensitivity in 3D culture. Probenecid itself inhibited the viability of 22Rv1 spheroids andsuppressed spheroid compaction rather than growth inhibition in the spheroids of another prostate cancer cellline PC-3. In addition, probenecid inhibited colony formation of 22Rv1 and PC-3 cells in soft agar anddecreased the protein levels of focal adhesion kinase (FAK), which is important for anchorage-independent
growth. In this presentation, we will also discuss about the latest findings including the target of probenecid inthese anti-cancer effects.

Copyright © The Japanese Pharmacological Society1-B-P-079

一般演題（ポスター）

Search for chromone derivatives that show high tumor-specificity against humanoral squamous cell carcinoma, and evaluation of their adverse effects on normalcells〇坂上 宏1、田沼 靖一1、天野 滋1、魚田 慎1、植沢 芳広2、黒崎 宏太2、高尾 浩一3、杉田 義昭3
明海大、2明治薬科大・医療分子解析学、3城西大・薬1

Many anticancer drugs have been reported to cause serious side effects such as oral mucositis,
neurotoxicity, and extravascular leakage. We have reported that among 291 chromone derivatives, 7-methoxy-3[(1 )-2-phenylethenyl]-4 -1-benzopyran-4-one (compound A) showed the highest tumor-specificity against
human oral squamous cell carcinoma (OSCC) cell lines, exceeding that of doxorubicin and 5-FU. In this study,
newly synthesized 65 chromone derivatives were investigated for their TS and side effects.

The 50%cytotoxic concentration (CC50) for four OSCC (Ca9-22, HSC-2, HSC-3, HSC-4), normal oral cells (gingival
fibroblast, periodontal ligament fibroblast, pulp cell), oral epithelial cells (HOK, HGEP) and differentiated
PC12 neuronal cells was determined from the dose-response curve. TS was calculated as the ratio of the mean
CC50 for normal cells to that for OSCC. Apoptosis was assayed by cell cycle analysis.
Newly synthesized indolychromones，indole-aurone hybrids，capsaicin derivatives，6,7-styrylchromones，3benzylidenechromanones showed much lower TS value than compound A. Compound A should much lower
keratinocyte toxicity than doxorubicin and 5-FU. 20 h treatment of Ca9-22 with compound A induced plateaulevel of cytotoxicity and accumulation of subG1 and G2/M population.

study suggests the inhibition ofcompound A against the estrogen related receptor-alpha signaling pathway, that is identified as an adverse
marker for breast cancer progression and poor prognosis.

Copyright © The Japanese Pharmacological Society1-B-P-080

一般演題（ポスター）

Canagliflozin, a sodium-glucose transporter 2 (SGLT2) inhibitor, suppresses thegrowth of glioblastoma through the AMPK-mTOR signaling〇江田 岳誉1、棗田 学2、大石 誠2、藤井 幸彦2、武井 延之3
新潟大・病院薬剤部、2新潟大・脳研・脳神経外科学、3新潟大・脳研・腫瘍病態学

1

Glucose is a major metabolic source required for cancer cell survival and growth. The up-take of glucose by its
transporters is enhanced in cancer cells. Cellular glucose levels are sensed by 5′-AMP-activated protein kinase(AMPK). Upon glucose insufficiency, AMPK is activated and suppresses anabolic processes such as mammaliantarget of rapamycin (mTOR) system. mTOR is a key molecule for cellular growth and dysregulated activation isobserved in cancer cells.
Therefore, we focused on the sodium-glucose transporter 2 (SGLT2) in glioblastoma survival and growth,because its expression was observed in these cells. We examined the effects of pharmacological inhibition of
SGLT2 by canagliflozin. Canagliflozin reduced the growth of glioblastoma cell lines of human origin in a dosedependent manner. Canagliflozin enhanced the phosphorylation of AMPK and suppressed S6 protein andp70S6 kinase phosphorylation. Canagliflozin inhibited the protein synthesis evaluated by the SUnSET assay.
Canagliflozin inhibited the growth of glioblastoma also in xenograft model.
Canagliflozin activates AMPK and inhibits mTOR pathway thus inhibits the glioblastoma growth both in vitro
and in vivo.

Copyright © The Japanese Pharmacological Society1-B-P-081

一般演題（ポスター）

LAT1 plays a critical role in cell proliferation via CDK1 and CDK2 in cabazitaxelresistant prostate cancer cells〇梨井 隼菱1、坂本 信一1、斎藤 心平1,2、新井 隆之1、溝上 敦3、安西 尚彦2、金井 好克4、市川 智彦1
千葉大・院医・泌尿器科学、2千葉大・院医・薬理学、3金沢大・院医・泌尿器科学、4大阪大・院医・生体システム薬理学

1

L-type amino acid transporter 1 (LAT1) is known to be highly expressed in various cancer types.
We explored the role of LAT1 in cabazitaxel-resistant prostate cancer cells using phosphoproteome analysis.
We used PC-3, and a cabazitaxel-resistant strain generated based on PC-3 (PC-3TxR/CxR). JPH203, a specific inhibitor of LAT1, was used to inhibit LAT1 function. Phosphoproteome analysis
was used to quantitatively investigate the proteins and sites of phosphorylation that are altered by JPH203
administration.
Compared to PC-3, LAT1 expression was significantly upregulated in PC-3-TxR/CxR. JPH203
significantly inhibited the migration and invasion of PC-3-TxR/CxR cell. Phosphoproteome analysis showedthat JPH203 treatment reduced the activity of CDK1 and CDK2 as kinases more than previously known mTOR
in PC-3-TxR/CxR cell. The decrease of phosphorylation in Cdc6 and Rb, substrates of CDK1 and CDK2,
respectively, by treatment with JPH203 was confirmed by Western blotting.
Current date may indicate that LAT1 has a crucial role to progression of cabazitaxel-resistant
prostate cancer via CDK1 and CDK2.

Copyright © The Japanese Pharmacological Society1-B-P-082

一般演題（ポスター）

Potential of lysophospholipids in the prevention and treatment of Alzheimer's
disease〇上芝 洸貴1、上田 勝也2、馬 闖2、泉谷 惇2、塚原 完3、松田 佳和4、齋藤 直人5、羽二生 久夫1,2,5
信州大・院総合理工、2信州大・院医理工、3長崎大・院医歯薬、4日本薬科大・薬、5信州大・バイオメディカル研究所

1

Alzheimer's disease (AD) is a disorder in which cognitive dysfunction appears due to neuronal damage caused
by abnormal aggregation of amyloid-beta protein (Aβ) in the brain. The number of patients is increasing year
by year, not only in Japan but also around the world. However, all drugs currently approved in Japan for thetreatment of AD are symptomatic treatments and cannot fundamentally cure the condition. Therefore,
establishing methods of prevention and therapeutic of AD is an important issue. The authors have shown thatporcine liver decomposition product (PLDP) exhibit a variety of biological activities in living organisms. In thisstudy, we evaluated the effect of PEL, lipids extracted from PLDP, on the aggregation process of Aβ, a key
protein in AD. PEL was extracted from PLDP by the Bligh & Dyer method. The thioflavin T assay was used as
the evaluation method to monitor changes over time in fluorescence caused by Aβ aggregation. The results
revealed that PEL dissociates as Aβ aggregates. In addition, the thioflavin T assay was also performed on
various lysophospholipids, which are abundant in PEL. The results confirmed the phenomenon of inhibition ofAβ aggregation in certain molecular species. These results suggest that PEL could be a first step in thedevelopment of AD drug discovery.

Copyright © The Japanese Pharmacological Society1-B-P-083

一般演題（ポスター）

Effects of 3,3‘-diindolylmethane, a phytochemical in Brassica vegetables, on theactivation of cardiac fibroblasts.

〇斉藤 麻希
医療創生大・薬・医療薬学

Phytochemicals are bioactive substances produced by plants so as to protect them from infection by fungi,
bacteria, and viruses. Nowadays, many scientists have focused on and reported their beneficial effects against
the activation of inflammatory cells and/or various types of cancer cells. In the present study, I investigated theeffects of 3,3ʻ-diindolylmethane (DIM), an indole derivative contained in Brassica vegetables, on the activationof cardiac fibroblasts, which is deeply involved in cardiac fibrosis and furthermore it would cause heart failure.
Human cardiac fibroblasts (hCF) were purchased and grew on collagen type I-coated culture dishes in themedium for hCF. The hCF were stimulated with 0.03~3 micro M of angiotensin II for 24 hrs present or absent
of various concentrations of DIM. After the stimulation periods, the total RNA of the cells was conventionally
extracted and then used for reverse transcription followed by real-time PCR.
The stimulation with angiotensin II tended to increase the expression of connective tissue growth factor(CTGF), one of the indicators for fibroblast activation, and concomitant application with DIM seemed tosuppress these increases. The expression levels of other molecules which seemed to be involved in the activationof the fibroblast, we investigated, had a similar tendency as CTGF. These results suggest that DIM has thepotential to be a candidate compound for the alleviation of the progression of cardiac fibrosis.

Copyright © The Japanese Pharmacological Society1-B-P-084

一般演題（ポスター）

Discovery of hypnotic effect of two Japanese traditional herbal medicines on
Drosophila insomnia models by using a newly developed automated sleep andrhythm analysis system (AutoCircaS)〇井上 栄二1,2、鈴木 孝洋2,3、清水 康晴1、岩城 良和4、川崎 陽久2、石田 直理雄2
1

救心製薬東京研、2国際科学振興財団 時間生物学研究所、3シグレイ、4タイセー【Purpose】 Sleep inwas defined in 2000 by using an equipment named Drosophila Activity(fly) to analyze theirMonitor (DAM) system. However, this DAM system has too narrow space forsocial behavior. To overcome such demerits of DAM system, we developed a novel automated sleep and rhythm
analysis system (AutoCircaS) which can monitor and record any behaviors like social mating, sleep, andcircadian rhythm in flies in free space using the time-lapse (one frame per 10 sec) imaging. This study aimed toinvestigate the hypnotic effect of Japanese traditional herbal medicines on fly insomnia models by using the
AutoCircaS.
【Method】 The caﬀeine-induced insomnia wild-type ﬂies and the short-sleep mutant [used to assess the hypnotic effects of Japanese traditional herbal medicines()] flies were

. Flies were placed

individually into 24-well microplates with drug-containing medium and acclimated for 3 days. The sleep
duration was measured for 3 days by using the AutoCircaS.
【Results and Discussion】 Shortening of the sleeping duration was recorded in the caﬀeine-induced wild-type
and the-mutant flies using AutoCircaS. In this new system, we found Japanese traditional herbal medicines,

KKG, NK and a Kampo hypnotic prescription Sansoninto, significantly improved the shortening of the sleep
duration in the caffeine-induced insomnia flies. Moreover, KKG also significantly improved the decrement of
the sleep duration in the-mutant insomnia flies. The data provide new insights into the use of KKG and NK

for human insomnia.

Copyright © The Japanese Pharmacological Society1-B-P-085

一般演題（ポスター）

Elucidation of improving effects of ninjinyoeito, a traditional Japanese herbal
medicine, on Aβ25-35-induced neurite damage〇窪田 香織、永松 拓海、岡村 尚幸、坂本 真由、石田 恵理奈、渡辺 拓也、桂林 秀太郎、岩崎 克典
福岡大・薬Ninjinyoeito (NYT), a Japanese herbal medicine, is widely used to treat patients with insomnia, anemia,
amnesia, and neurosis. Recently, NYT was reported to be clinically effective in Alzheimer's disease (AD). To
address the mechanism underlying effect of NYT on AD, we examined the effects of NYT in β-amyloid (Aβ)25‒-exposed primary cocultured astrocytes and neurons. The effects of NYT on neurotoxicity induced by Aβ25‒35

35

were assessed by immunocytochemical assays and Sholl analysis of microtubule-associated protein 2 (MAP2)positive and tau-positive neurites. Aβ25‒35 treatment attenuated the arborization of axons and dendrites of single
autaptic hippocampal neurons in a concentration-dependent manner. NYT treatment ameliorated the Aβ25‒35induced impairment of tau-positive axon outgrowth. However, NYT did not ameliorate the Aβ25‒35-induced
suppression of MAP2-positive dendrite arborization. RT-PCR analysis demonstrated that NYT increases theexpression of nerve growth factor (Ngf) but not brain derived neurotrophic factor (Bdnf) in Aβ25‒35-exposed
primary cocultured astrocytes and neurons. Our results indicate that NYT protects against Aβ25‒35-induced
neuronal injury through induction of nerve growth factor expression. These findings provide a mechanistic basis
for treating AD with NYT.

Copyright © The Japanese Pharmacological Society1-B-P-086

一般演題（ポスター）

Action of Rikkunshi-To on the rivastigmine-induced nausea in mice

〇山本 浩一1、佐藤 雄己2
1

森ノ宮医療大・医療技術・診療放射線、2福山大・薬・臨床薬効解析

Rivastigmine, which is a cholinesterase inhibitor used for the treatment of mild to moderate Alzheimer's disease,is known to be associated with significant gastrointestinal (GI) adverse reactions including nausea, vomiting,anorexia. If severe GI adverse reactions are observed during the course of treatment, the patient should be
instructed to stop treatment. Rikkunshi-To (RKT), a traditional herbal Japanese medicine, has been prescribedfor patients with various GI symptoms, and we have reported that RKT on therapy-induced nausea in mice. We
also reported that pica, kaolin ingestion behavior, could be used to evaluate nausea in mice; thus, in this study,
we investigated the effects of rivastigmine on pica in mice and the effects of RKT on inhibition of rivastigmineinduced nausea.
Male mice were consecutively administered rivastigmine (i.p.) for 5 days, and their kaolin intakes were
measured. Additionally, we examined the effects of a serotonin 5-HT3 (granisetron: i.p.), dopamine D2(domperidone: i.p.) and muscarinic (butylscopolamine: i.p.) receptor antagonists rivastigmine-induced pica.
Furthermore, we investigated whether RKT has the therapeutic effects on the rivastigmine-induced pica.
We found mice showed pica during the course of donepezil administration. Among the tested anti-emetic drugs,
none of the anti-emetic drugs led to any change in the rivastigmine-induced pica. On the other hand, mice fed
the diet supplemented with RKT significantly inhibited rivastigmine-induced pica. These findings suggest that
RKT is useful to prevent rivastigmine-induced nausea.

Copyright © The Japanese Pharmacological Society1-B-P-087

一般演題（ポスター）

Effect of lactic acid bacteria-fermented milk whey on melanin production〇五十嵐 信智1、西中 ゆい1、篠﨑 優衣1、吉田 涼太郎1、田端 慶斗1、今 理紗子1、酒井 寛泰1、畑中 美咲2、細江 智夫
11

星薬科大、2アサヒグループ食品

Melanin is generated in the melanosomes of melanocytes and is an important factor determining the skin color.
Melanogenesis is catalyzed by tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1), and dopachrome
tautomerase (DCT). Therefore, these enzymes are suitable targets for the development of cosmetics aimed atskin whitening. In this study, we investigated the effect of milk whey on melanin production. Whey obtained
from five strains of lactic acid bacteria (L. delbrueckii bulgaricus, L. helveticus, L. acidphillus, L. casei, and L.
helveticus CM4 strains) was used. The melanin contents in B16 melanoma cells treated with α-MSH wassignificantly increased by about 2-fold compared to control cells. In contrast, melanin production by α-MSH
was significantly suppressed in whey obtained from any lactic acid bacteria. These wheys inhibited α-MSHinduced increased expression of TYR, TYRP1, and DCT. When the whey obtained from L. helveticus was
examined in detail, it was found that the fraction with a molecular weight of 3 kDa or more had a strong melanin
production inhibitory effect. These results demonstrated that whey decreases the expression of TYR, TYRP1,
and DCT and suppresses melanin production. It was also suggested that substances with a molecular weight of 3
kDa or more may be involved in this action of whey.

Copyright © The Japanese Pharmacological Society2-B-P-088

一般演題（ポスター）

Alteration of stress granule formation and clearance by S-Nitrosylation
modification of G3BP1〇伊藤 和、藤河 香奈、上原 孝
岡山大・院医歯薬・薬効解析学

Nitric oxide (NO) is a signaling molecule that exerts a variety of regulatory functions in physiological states andstress responses. NO has been proposed to modulate protein function through -nitrosylation of cysteine thiol
residues in proteins. Appropriate amounts of NO result in neuroprotective effects via increasing moderate nitrosylated proteins and the cGMP pathway. On the other hand, excess amounts of NO promote
neurodegenerative signaling pathways by increasing aberrant -nitrosylated proteins.
We found that Ras GTPase-activating protein-binding protein 1 (G3BP1), which forms stress granules (SGs)
implicated in neurodegenerative diseases, was a target of -nitrosylation. Treatment with NO stimulated thismodification in a concentration-dependent manner. It has been known that SGs are rapidly formed by oxidativeand heat shock stresses. In addition, SGs formation induced by proteasome inhibitor (MG132) was graduallydegraded, whereas NO delayed this degradation. These findings suggest that -nitrosylated G3BP1 not only
promotes SGs formation but also delays SGs clearance.

Copyright © The Japanese Pharmacological Society2-B-P-089

一般演題（ポスター）

Ghrelin restores the dopamine responses in the PFC to external stimuli via
dopamine D1 receptor signaling and attenuates cognitive deficit in MECP2 KO
mice, a model mouse for Rett syndrome〇河原 幸江1、大西 克典1、高橋 知之2,3、岸川 由紀1,4、弓削 康太朗2,3、河原 博5、山下 裕史郎2,3、松石 豊次郎2,6、西 昭
徳1
1

久留米大・医・薬理、2久留米大・高次脳疾患研、3久留米大・医・小児科、4西九州大・リハビリテーション、5鶴見大・歯・歯科麻酔、6聖マリア病院・小児総合研究セ・レット症候群研究セGhrelin improves cognition in various animal models with cognitive deficits. This study investigated the effects
of ghrelin on cognitive deficits and modulation of D1 receptor signaling in the PFC, which plays a critical role incognitive performance, inknockout (KO) mice using

microdialysis.

In the modified novel object recognition (NOR) test, KO mice showed the impairment of cognition. Ghrelin
injection (8.6 microgram/mouse, s.c.) improved the cognition of objects and investigatory behaviors in KO
mice. In microdialysis studies, saline injection and novelty induced increases in DA in WT mice, and theincrease was not observed in KO mice. Thus, KO mice exhibit the low response of DA to external stimuli in the
PFC. The results of the infusions of D1 receptor ligands into the PFC indicate that D1 receptor signaling in WT
mice is involved in bidirectional regulatory mechanisms of DA release. In KO mice, the ability of D1 receptor
signaling to inhibit DA release would be upregulated under tonic and D1 receptor-stimulated conditions.
Ghrelin injection, but not saline injection, increased DA levels in the PFC, and the DA levels significantly
increased in response to the novelty after ghrelin injection in KO mice. The DA responses to the ghrelin
injection and to the novelty after ghrelin injection in KO mice were completely abolished by the infusion of
SCH23390.
Ghrelin restores DA responses to external stimuli by adjusting the altered function of D1 receptor signaling, andthe action of ghrelin may underlie the mechanism for ameliorating cognitive deficit in

Copyright © The Japanese Pharmacological Society

KO mice.

2-B-P-090

一般演題（ポスター）

Role of the microsomal prostaglandin E synthase-1 in imiquimod-induced
psoriasis-like skin inflammation.

〇日置 優花1,2、榎本 大樹1、三浦 早貴1、小野寺 優1、板橋 輝1、飯塚 佳子3,4、前花 祥太郎3,5、久保 誠3,5、北里 英朗6、
市川 尊文2,3、小島 史章1,2,3
北里大・医療衛生・薬理学、2北里大・院医療・生体制御生化学、3北里大・医療衛生学部附属・再生医療・細胞デザイ

1

ン研究施設、4北里大・院医療・食予防医科学、5北里大・院医療・環境微生物学、6北里柴三郎記念館

Psoriasis is a chronic inflammatory disease that is accompanied by abnormalities in the immune
system. It was recently reported that microsomal prostaglandin E synthase-1 (mPGES-1), a terminal enzyme for
PGE2 biosynthesis, highly expresses in the skin of psoriasis patients. However, the detailed role of mPGES-1 inpsoriasis remains unclear. In this study, we investigated the role of mPGES-1 in psoriasis-like skin inflammationinduced by imiquimod (IMQ), one of the well-established models of psoriasis.
Psoriasis-like skin inflammation was induced in mice lacking mPGES-1 (mPGES-1−/− mice) andwild-type (WT) mice by administrating IMQ under specific pathogen free condition. The expressions of
mPGES-1 mRNA and protein in the skin were determined by real-time PCR and western blotting, respectively.
The skin inflammation was evaluated based on scores with macroscopic symptoms and histological features. In
addition, the expression levels of intereukin-17A (IL-17A) in inflamed skin was determined by real-time PCR.
The expression of mPGES-1 was highly induced on both mRNA and protein levels in the skin of WT
mice after IMQ administration. Interestingly, the mPGES-1−/− mice exhibited more severe symptoms ofpsoriasis-like skin inflammation compared to those of WT mice during administration of IMQ. Histologicalanalysis further showed significant increase of epidermal thickness in mPGES-1−/− mice. The skin expression of
IL-17A, a prominent target for the treatment of psoriasis, was highly up-regulated in mPGES-1−/− mice in
response to IMQ administration.
mPGES-1/PGE2 system plays a protective role in psoriasis, partly by reducing the expression of IL-17A.

Copyright © The Japanese Pharmacological Society2-B-P-091

一般演題（ポスター）

Thromboxane A2 receptor signaling inhibits angiogenesis andlymphangiogenesis in the endometriotic lesions in mice

〇古江 明子1,2、伊藤 義也1、本田 雅子2、服部 響子2、関口 和企2、山下 敦1、長田 真由子1、田邊 美奈1、細野 加奈子
1、畑中 公1、馬嶋 正隆3、加藤 一喜2、天野 英樹11

北里大・院医療・分子薬理、2北里大・医・産婦人科、3神奈川工科大学・健康医療科学部・病態治療

Angiogenesis and lymphangiogenesis contribute to the development of endometriosis. We recently reported thatthromboxane A2 (TXA2) receptor signaling involves in angiogenesis in critical limb ischemia andlymphangiogenesis in inflamed diaphragm. In the present study, using wild-type mice (WT) and thromboxane
prostanoid receptor (TP) knockout mice (TPKO), we examined whether TP signaling plays a role in the growthof endometriosis by angiogenic responses. Ectopic endometriosis model was created by transplantation ofendometrial tissue fragments from donor mice (WT or TPKO) into the peritoneal wall of host mice (WT or
TPKO). The implant sizes and density of blood and lymphatic vessels in the TPKO implants from host TPKO
(TPKO→TPKO) were increased as compared with the WT→WT. The mRNA levels of markers for blood(CD31) and lymphatic vessels (LYVE-1) and of growth factors for angiogenesis (VEGF-A) andlymphangiogenesis (VEGF-C/D) in the TPKO→TPKO were higher than those in the WT→WT.
Immunostaining showed that TP was expressed in F4/80-positive macrophages, but not in blood and lymphatic
vessels in endometriosis lesions. The levels of M2 macrophage-related genes were higher in the TPKO→TPKO
than in the WT→WT, while no statistically significant difference in M1 macrophage-related genes was observed
These results suggest that TP signaling inhibits the growth of endometriosis by reducing angiogenesis andlymphangiogenesis.

Copyright © The Japanese Pharmacological Society2-B-P-092

一般演題（ポスター）

Anorexigenic effects of central administered xenin was possibly induced via
central nesfatin-1 cells in rats

〇齊藤 将太、橋本 弘史、濱口 紀江、裵 祥存、齋藤 心平、霊園 良恵、平山 友里、安西 尚彦
千葉大・院医・薬理学教室

Xenin, which was identified in human gastric mucosa, is a 25-amino acid peptide. Xenin is widely expressed inperipheral and central tissues. Xenin has various physiological functions, such as stimulating intestinal motility.
Central and peripherally administered xenin decreased food intake in rodents. Nesfatin-1/NUCB2 (nesfatin-1),which is an anorexic neuropeptide consisting of 82 amino acids, is widely expressed in peripheral and central
tissues. This study examined the effect of intraventricular (icv) administration of xenin on central nesfatin-1
cells in rats. Fos immunohistochemistry was used to investigate the effect of icv administration of xenin on
nesfain-1-like immunoreactive (LI) cells in rat brain. Fos-LI cells were observed in the supraoptic nucleus,
paraventricular nucleus, arcuate nucleus, and nucleus of the solitary tract after icv administration of xenin.
Double immunohistochemistry for Fos and nesfatin-1 showed that nesfatin-1-LI cells expressing Fos weresignificantly increased compared with a control group in these nuclei after icv administration of xenin.
Anorexigenic effects of xenin were attenuated by nesfain-1 antisense pretreatment. These results suggested thatthe anorexigenic effect of xenin is partially mediated by nesfatin-1 cells in rats.

Copyright © The Japanese Pharmacological Society2-B-P-093

一般演題（ポスター）

BQ788, a selective ETB receptor antagonist alleviates inflammatory reactions
after traumatic brain injury in mice

〇道永 昌太郎1、水口 博之2、小川 泰弘1、菱沼 滋1、小山 豊3
明治薬大・薬・薬効、2大阪大谷大・薬・薬理、3神戸薬大・薬・薬理1Traumatic brain injury (TBI) is a fetal damage to the brain resulted from an external force to head by accidents
and falls. One of the TBI-induced severe pathogenesis is an inflammatory damage. We previously suggested that
BQ788, an endothelin ETB receptor antagonist alleviated blood-brain barrier disruption and brain edema in TBI
mice. In this study, we investigated the effects of BQ788 on inflammatory reactions in TBI mice. As a model of
TBI, a fluid percussion injury (FPI) was performed by a hydraulic impact on the mouse dura mater. BQ788 (15nmol/day) was repeatedly administrated into lateral cerebroventricle from 2 to 5 days after FPI. As a maker ofneutrophil, myeloperoxidase (MPO) and Ly6G were examined by fluorescent immunostaining. Expressions ofinflammatory cytokines (tumor necrosis factor-α: TNF-α and interleukin-1β: IL-1β) and chemokines(monocyte chemoattractant protein−1: MCP−1 and mouse macrophage inflammatory protein-2: MIP-2) were
measured by Real-time PCR. After FPI, MPO- and Ly6G-posotive cells were increased in the mouse cerebrum.
Administration of BQ788 decreased these positive cells. Additionally, BQ788 decreased FPI-induced increases
in expressions of TNF-α, IL-1β, MCP-1 and MIP-2 in the mouse cerebrum. These results suggest that ETB
receptor antagonist alleviates TBI-induced inflammatory reactions.

Copyright © The Japanese Pharmacological Society2-B-P-094

一般演題（ポスター）

cGAMP-induced metabolic alterations in astrocytes and their impacts on tumor
immune responses in the CNS〇佐藤 洋美1、菊池 望恵1、後藤 杏子1、大川 柊弥1、松本 千佳2、田中 浩揮2、秋田 英万3、樋坂 章博1
千葉大・院薬、2千葉大・院薬・薬物学、3東北大・院薬・薬物送達学分野

1

It has been shown that cGAMP (Cyclic 2'3'-GMP-AMP) is delivered via gap junction from cancer cells to
adjacent astrocytes in metastatic brain tumors (BrM) and contributes to tumor progression. We aimed toreproduce the peritumoral astrocytic conditions by introducing cGAMP directly into astrocytes and to quantify
the metabolic changes around the glutamine (Gln)-glutamate (Glu) cycle, as major CNS metabolism. In
cGAMP-treated cells, the synthetic flux of Gln from 13C5,15N-Glu was reduced to about half that of control cells,
while Glu secretory flux was increased two-fold. Furthermore, when Glu was evaluated as a chemoattractant for
migration using HL60-derived neutrophils, which have been implicated in immunosuppression of BrM,
exposure to 10-50 μM Glu tended to increase the migration. Downstream of the STING, which accepts
cGAMP, a marked upregulation was observed for IL-6, CCL5, CCL2, CCL7, and CXCL1 by several tens of fold.
They are target genes of the NF-κB pathway that may be involved in interferon activation as well as IRF3, andwere completely suppressed by NF-κB blockage. Glu-induced oxidative stress has been reported to mediate
activation of NF-κB. These results suggest that induction of NF-κB by cGAMP may be involved in Glu
metabolic alterations, as well as in tumor microenvironment formation, including immune responses.

Copyright © The Japanese Pharmacological Society2-B-P-095

一般演題（ポスター）

Possible involvement of monomeric forms of visfatin around the cerebral bloodvessels in the pathogenesis of cerebral infarction with diabetes〇岩谷 結衣、山本 春菜、市川 美月、増田 紋、森山 慶之、林 秀樹、高木 教夫
東京薬科大・薬・応用生化学

Changes in visfatin, as an adipocytokine, after cerebral infarction with diabetes and its pathophysiologicalsignificance are not clear. We attempted to elucidate the relationship between changes in visfatin after cerebral
infarction with diabetes and pathogenesis. Protein levels of visfatin in brain tissue after middle cerebral artery
occlusion/reperfusion (MCAO/R) in db/db mice did not change compared with those of the non-diabetic andcontrol groups, but they were significantly increased in the cerebrovascular fraction. This increase would bederived from astrocytes as a monomer, which is involved in inflammation, around the cerebral blood vessels.
The increase in MMP-9 mRNA of the cerebrovascular fraction might be due to the increased monomeric formof visfatin. Interestingly, there were no changes in visfatin mRNA in the cerebrovascular fraction, while protein
levels of dimeric form of visfatin, which contributes to neuroprotection, were increased in serum after MCAO/R
with diabetes. Therefore, visfatin could be secreted into the blood from peripheral tissues as a dimer after
MCAO/R with diabetes, and a part of it changes into a monomer in the process of infiltration into the brain
parenchyma. Monomeric forms of visfatin may be involved in an inflammatory reaction and the pathogenesis of
cerebral infarction with diabetes.

Copyright © The Japanese Pharmacological Society2-B-P-096

一般演題（ポスター）

Upregulation of nicotinic acetylcholine receptors in pulmonary arterial
hypertension〇山村 彩1、中浜 光哉2、Alamgir Hossain1、北村 文也3、高橋 理恵1、山村 寿男2、佐藤 元彦1
愛知医科大・医、2名古屋市大・院薬・細胞分子薬効解析学、3愛知医科大・医・腎臓リウマチ膠原病内科

1

Smoking causes hypoxic vasospasm, thickening, and inflammation. Such responses are similar to thepathological mechanism of pulmonary arterial hypertension (PAH). PAH is a progressive and fatal disease thatis characterized by the irreversible remodeling of the pulmonary artery. Pulmonary vasospasm, thickening, andinflammation are triggered by a chronic increase in cytosolic Ca2+ concentration. Here, we focused on theexpression of nicotinic acetylcholine receptors (nAChRs), which are associated with cytosolic Ca2+ signaling, in
PAH and hypoxic stress. The expression of the α subunits of nAChRs in pulmonary arterial smooth muscle cells(PASMCs) from normal subjects and idiopathic pulmonary arterial hypertension (IPAH) patients was analyzed
by RT-PCR. Normal-PASMCs expressed nAChRα5 and α9 subunits. On the other hand, IPAH-PASMCs
expressed nAChRα1, α5, and α7 subunits. As a result of Western blotting, the expression of nAChRα1andα7 proteins was upregulated in IPAH-PASMCs. In addition, the expression of nAChRα1 subunits was also
increased in PASMCs from monocrotaline-induced pulmonary hypertensive rats. Furthermore, hypoxic
exposure (1% O2) increased nAChRα7 expression in normal-PASMCs. In conclusion, the expression ofnAChRα1 and α7 subunits is upregulated in chronic respiratory diseases including PAH and hypoxic stress.

Copyright © The Japanese Pharmacological Society2-B-P-097

一般演題（ポスター）+

2+

Involvement of mitochondrial Na /Ca exchanger in hypoxia-induced pulmonary
arterial hypertension〇喜多 紗斗美1、田頭 秀章2、披田 真里1、谷 和佳奈1、根本 隆行2、喜多 知2、岩本 隆宏2
徳島文理大・薬・薬理、2福岡大・医・薬理1

Pulmonary arterial hypertension (PAH) is a severe and progressive disease that leads to right heart failure. The
pathogenesis of PAH is generally characterized by vasoconstriction, upregulated proliferation, migration, andpulmonary vascular remodeling in lung tissue. Recent studies using genetic analyses and experimental models
have suggested that the hypercontraction of pulmonary arteries induced by Ca2+ signaling abnormality may be
involved in the pathogenesis of PAH. The Na+/Ca2+ exchanger type-1 (NCX1) is a bidirectional transporter thatis controlled by membrane potential and transmembrane gradients of Na+ and Ca2+. We recently showed that the
upregulation of vascular smooth muscle NCX1 contributes to the development of hypoxia-induced PAH, using
NCX1 knockout mice and specific NCX1 inhibitor SEA0400. In the present study, we investigated thepathological role of mitochondrial Na+/Ca2+ exchanger (NCLX). NCLX knockout mice exhibited significant
reduction in right ventricular systolic pressure compared with wild-type mice. Furthermore, specific NCLX
inhibitor CGP-37157 significantly suppressed hypoxia-induced PAH in wild-type mice. These findings indicatethat NCLX contributes to the development of hypoxia-induced PAH, suggesting that NCLX inhibition might be
a novel approach for the treatment of PAH.

Copyright © The Japanese Pharmacological Society2-B-P-098

一般演題（ポスター）

Involvement of xenobiotic efflux transporter MRP5/ABCC5 in neurite outgrowth〇石本 尚大、八木 寛史、増尾 友佑、加藤 将夫
金沢大・薬学系

Multidrug resistance associated protein 5 (Mrp5/Abcc5) is highly expressed in mouse primary cultured neurons(PCN) compared to other Abcc family members. However, the physiological role of Mrp5 in neurons is still
unknown. In the present study, we examined the involvement of Mrp5 in neurite outgrowth because its
physiological substrates cGMP and cAMP are important modulators of the neurite outgrowth.
Immunocytochemical analysis showed that Mrp5 was expressed in the PCN and neuronal cell line Neuro2a.
Efflux activity of Mrp5 in Neuro2a was then examined using CMFDA, which is metabolized to a fluorescent
substrate for Mrps after diffusion into the cells. Knockdown of Mrp5 by transfection with siRNA for Mrp5
(siMrp5) showed a higher fluorescence intensity during incubation with CMFDA compared with control siRNA.
In addition, exposure to an Mrp5 inhibitor zaprinast increased the fluorescent intensity. These results show thefunctional expression of Mrp5 in Neuro2a. Furthermore, both siMrp5 and zaprinast significantly increased
number of cells having longer neurites. The knockdown with siMrp5 tended to increase protein expression ofcGMP-dependent protein kinase 1 (PKG1), while a PKG inhibitor significantly suppressed the neurite
outgrowth induced by Mrp5 knockdown with a concomitant decrease in expression of PKG1, suggesting that
inhibition of MRP5 would promote neurite outgrowth at least partially through activation of PKG1. Further
studies are needed to elucidate the possible involvement of clinically used Mrp5 inhibitor drugs in the neurite
outgrowth.

Copyright © The Japanese Pharmacological Society2-B-P-099

一般演題（ポスター）

Intragingival application of Porphyromonas gingivalis-derived
lipopolysaccharide induces an increase in plasma TNF-α levels in anaesthetisedrats〇青野 悠里、斉藤 幸治、三枝 禎
日本大・松戸歯・薬理Periodontal disease is a chronic inflammation of the gingiva resulting in the destruction of periodontal tissue.
(), a gram-negative bacterium, appears to play a role in the development of

periodontal disease. Lipopolysaccharide (LPS), a component of the gram-negative bacterial cell wall, plays arole in periodontal tissue destruction. We have shown that intragingival application of-derived LPS (-LPS)

in rats increases gingival TNF-α without affecting IL-6 (Taguchi et al., Int. J. Oral Sci., 2015). Since
periodontal infection is known to be a potential threat to general health, we examined the influence of
intragingival application of-LPS on plasma TNF-α and IL-6 levels in urethane-anaesthetized rats. For

comparison, effects of LPS derived from(-LPS) were also studied. Male SD rats were used.

The external jugular vein was cannulated and connected via Teflon tubing to a blood sampling system. Either-LPS (1 µg),

-LPS (6 µg) or vehicle was injected into gingiva of upper incisors via a microsyringe, with 500µl of blood sampled every 1 hr. Plasma TNF-α and IL-6 levels were determined by enzyme-linked assays.
Unlike IL-6, basal plasma TNF-α was lower than the detection limit (5 pg/ml).
levels but increased TNF-α. Neither IL-6 nor TNF-α were altered byincreased-LPS failed to affect IL-6

-LPS. These results suggest that

-LPS in gingiva may have systemic effects through induction of plasma TNF-α.

Copyright © The Japanese Pharmacological Society2-B-P-100

一般演題（ポスター）

ATP release from astrocyte induced by mechanical stimulation and its
visualization〇加藤 奥穂、井上 貴文
早稲田大・先進研・生医 井上研

ATP, a gliotransmitter released from astrocytes, works in glia-glia and glia-neuron communications. Still, thefunctional significance of astrocyte-derived ATP remains to be elucidated, since there has been no efficient andselective way to inhibit the ATP release. While two types of ATP release from astrocytes are known: vesicularand channel-mediated ATP release, the mechanisms of the channel-mediated ATP release are still unknown. To
investigate those, we established a method that can directly detect single ATP release events from single
astrocytes: ATP release was evoked by pressing cultured astrocytes with a glass pipette (mechanicalstimulation), and released extracellular ATP was detected using GRAB-ATP1.0, a protein ATP sensor together
with the changes in intracellular Ca2+ with a Ca2+ indicator, Fura-2. Extracellular ATP and intracellular Ca2+ wereobserved to increase in synchrony. Through pharmacological screening with inhibitors, maxi-anion channels,
connexin hemichannels, and volume-regulated anion channels revealed as candidates involved in astrocytic ATP
release. Further analysis will provide detailed mechanisms and functions of ATP release from astrocytes.

Copyright © The Japanese Pharmacological Society2-B-P-101

一般演題（ポスター）

Bitter taste receptor14 regulates expression of histamine receptors, E-Cadherin
and N-Cadherin〇小笠原 正人1、福田 尚代2、石河 太知3、徳弘 圭造2、加茂 政晴4、山田 浩之5、石崎 明4岩手医科大・歯・薬理、2関西医科大・生命科学・ゲノム編集、3岩手医科大・歯・微生物、4岩手医科大・歯・生化学、5岩

1

手医科大・歯・口腔外科

Background
Bitter taste receptor 14 belongs (TAS2R14) to GPCR family, stimulated by epigallocatechin gallate (EGCG).
We reported the EGCG administration induced downregulation of histamine receptors in oral epithelial cancer
cell lines. It is reported that usage of histamine receptor antagonists inhibited cancer cell metastasis throughsuppressing epithelial to mesenchymal transition (EMT). The current study investigates the function of
TAS2R14 for histamine receptors and EMT.
MethodWe performed knock out of TAS2R14 in HSC4 oral epithelial cancer cell line by genome editing method. The
cultured cells extractions were prepared for the western blot analysis. Histamine H1 and H2 receptor antibodies
were used for detection of protein expression. For the biochemical analysis of EMT, E-cadherin and N-cadherin
antibodies were used.
Results
Genome editing of TAS2R14 in three HSC4 cells resulted in integration of pretermination codon in the gene.
TAS2R14 protein in edited HSC4 cells was not detected by western blot analysis. E-cadherin was upregulated
and N-cadherin was down regulated. Furthermore, histamine H1 and H2 receptors were down regulated.
Conclusions and discussion
TAS2R14 is involved in regulation of EMT marker proteins and histamine H1 and H2 receptors expression.
EGCG administration to HSC4 cells means that there might be other molecular targets of EGCG or EGCG
might function as inverse agonist considering the reports that bitter taste receptors work as constitutive active
receptors.

Copyright © The Japanese Pharmacological Society2-B-P-102

一般演題（ポスター）+

Involvement of Kir2.1 K channel in osteoblast differentiation mediated by ATPconductive hemichannels〇鬼頭 宏彰、劉 澤成、雑賀 紀明、遠藤 京子、梶栗 潤子、大矢 進
名古屋市立大・院医

Osteoblast growth and differentiation, which are controlled by Ca2+ signaling, are important for bone formationand homeostasis. It has been reported that the inward rectifier Kir2.1K+ channel is responsible for osteoblast
differentiation by maintaining the resting membrane potential to regulate intracellular Ca2+ concentration ([Ca2+]i). ATP released from hemichannels, such as pannexins and connexins located within the cell membrane, also

contributes to osteoblast differentiation. The purpose of this study is to elucidate the functional relationshipbetween Kir2.1 and ATP signaling in osteoblast differentiation of the osteoblastic cell line MC3T3-E1,
established from mouse calvaria. We found that the alkaline phosphatase ALP mRNA expression in MC3T3-E1
cells and the endochondral ossification of metatarsal bones were suppressed by the treatment with ML133, aKir2.1 inhibitor or CBX, a hemichannel blocker. The pharmacological blockade of hemichannels or Kir2.1 also
decreased the amount of released ATP from MC3T3-E1 cells during osteoblast differentiation. ATP stimulationinduced a transient Ca2+ influx through ATP-sensitive Ca2+ permeable channels, and this ATP-stimulated [Ca2+]i
rises were attenuated by the treatment with ML133 and 5-BDBD, a P2X4 inhibitor. These results suggest thatboth Kir2.1 and hemichannels may be involved in ATP signaling during osteoblast differentiation. The
functional coupling between Kir2.1 and P2X4 channels plays an essential role in maintaining the bonehomeostasis via modulating osteoblast differentiation.

Copyright © The Japanese Pharmacological Society2-B-P-103

一般演題（ポスター）

Involvement of FAT/CD36 expression in mouse parotid ducts on salivary
secretion〇佐藤 慶太郎1、大野 雄太2、長瀬 春奈2、柏俣 正典2、安達 一典1
明海大・歯・薬理、2朝日大・歯・薬理1

Xerostomia is commonly observed in middle-aged and elderly patients. Since the adipose tissue infiltration isfrequently observed in the parotid gland (PG) of alder animals. We hypothesized that the alteration in
expression of fatty acid translocase (FAT/CD36), which facilitates fatty acids transportation, induces xerostomia
through the hyposecretion in the PG. We firstly examined theexpression in the three major salivary

glands of male BALB/c mice. The PG expressed significantly substantial

among them. In addition, theimmunohistochemical analysis revealed that the CD36 protein localized in duct cells, but not acinar cells, in the
PG of BALB/c mice. Then, the effect of CD36 inhibitor, sulfosuccinimidyl oleate, treatment on the salivary
secretion in 48-weeks BALB/c mice was assessed. The inhibitor reduced pilocarpine (Pilo)-induced salivation.
Moreover, the involvement of the PG CD36 in the salivary secretion of male 48-weeks senescence-accelerated
mouse (SAM) was investigated. Compared with SAM resistant 1 (SAMR1), the Pilo-induced salivation in agematched SAM prone 1 (SAMP1) was decreased. In addition, the protein expression of PG CD36 in SAMP1 wassignificantly lower than that of SAMR1. These results suggest that the CD36 in ducts of PG plays an importantrole in the aging-induced hyposecretion.

Copyright © The Japanese Pharmacological Society2-B-P-104

一般演題（ポスター）

Humanin promotes exocytosis in PC12 cells〇小塚 彩里、新倉 貴子
上智大・理工PC12 cells, rat pheochromocytoma cells, are neuroendocrine cells and widely used for the study of exocytosis.
PC12 cells release catecholamine (CA) from vesicles in response to the neurotransmitter acetylcholine (ACh) by
the fusion of the vesicle membrane and the cell membrane. In the models of neurodegenerative disorders, suchas Alzheimerʻs disease (AD), alteration of exocytosis mechanisms has been reported. Humanin (HN) is a 24residue polypeptide and initially identified as a neuroprotective factor against AD-related insults. HN
suppresses neuronal death caused by amyloid beta, an AD-associated cytotoxic insult, in vitro and ameliorates
memory deficit of AD mouse models. We have recently found that HN increases the levels of neurotransmitters
including catecholamines in the hippocampal region of normal mice. This finding suggests that HN may havethe direct effect on promoting exocytosis in neurons. In this study, we examined the effect of S14G-HN (HNG),
a highly potent HN derivative, on exocytosis. In PC12 cells, HNG increased ACh-induced CA secretion by the
Bath application experiment. On the other hand, neuroprotection-defective HN derivatives, S7A-HN and C8AHN, did not affect CA secretion. Furthermore, no secretagogue activity of HNG was observed by inhibiting JAK,a signal molecule of gp130 receptor. These results suggest that the receptor-mediated JAK-STAT pathway is
involved in the HNʻs activity for promoting exocytosis.

Copyright © The Japanese Pharmacological Society2-B-P-105

一般演題（ポスター）

MicroRNA targeting Skp1 regulates inflammatory response through NF-κB
signaling pathway〇佐野 朋美1、李 栄智2、溝上 顕子3、西村 英紀2、兼松 隆1
九州大・院歯・口腔機能分子科学、2九州大・院歯・歯周、3九州大・院歯・OBTセ

1

Nuclear factor-kappa B (NF-κB) is one of the transcription factors that play a central role in the immune
response and is involved in many physiological phenomena such as acute and chronic inflammatory reactions,cell proliferation and apoptosis. It has been reported that microRNA (miRNA) binds mainly to 3'UTR of thetarget mRNA, suppresses its gene expression, and plays an important role in various diseases such as cancer.
Therefore, the purpose of this study was to identify miRNA that regulates the activation of NF-κB signaling.
We identified miR-582-5p as a significantly downregulated miRNA in inflamed mouse adipose tissue. We foundmiR-582-5p was suppressed in LPS-stimulated RAW264.7 cells, a macrophage-like cell line. Subsequently, Sphase kinase-associated protein 1 (SKP1), a component of the E3 ubiquitin ligase that regulates the NF-κB
pathway, was detected as a potential target for miR-582-5p predicted by Targetscan. The dual luciferase
reporter assay confirmed that miR-582-5p binds to the 3'UTR site of, and the transfection of miR-582-5p

mimic suppresses SKP1 expression in RAW264.7 cells. In addition, miR-582-5p suppressed the production of
the inflammatory cytokine TNF-α by inhibiting the degradation of IκBα, which subsequently suppressed thephosphorylation of p65 resulting in its nuclear translocation. Our findings demonstrated that exogenously
applied miR-582-5p can attenuate the activation of NF-κB signaling pathway by targeting
expression, which provides a prospective therapeutic strategy for treating inflammatory diseases.

Copyright © The Japanese Pharmacological Societygene2-B-P-106

一般演題（ポスター）

The role of primary cilia in zebrafish fin regeneration.

〇白水 崇1、山川 大史2、稲垣 昌樹1、西村 有平11

三重大・院医・統合薬理、2三重大・院医・分子生理学

Primary cilia are immobile structures that extend from the surface of various cells and have been suggested to be
involved in signal transduction from extracellular stimuli. Signal transduction through primary cilia has beenreported to work in tissue regeneration, but the detailed mechanism remains unclear. Zebrafish has a higher
regenerative capacity than mammals and is a suitable animal model for observing the tissue regeneration
process. In this study, we investigated the role of primary cilia in tissue regeneration using knockout zebrafish ofgenes involved in the regulation of primary cilia formation. Trichoplein is localized in the basal bodies ofprimary cilia and works as a suppressor of primary cilia formation through activation of Aurora A kinase.
KCTD17 is also involved in primary cilia formation through the degradation of trichoplein. In the zebrafish fin
injury-repairing model, the trichoplein knockout fish showed higher regenerative capacity than the wild type. In
this presentation, we demonstrate the role of trichoplein and KCTD17 in the process of zebrafish finregeneration.

Copyright © The Japanese Pharmacological Society2-B-P-107

一般演題（ポスター）

Evaluating drug efficacy by visualizing the spatiotemporal dynamics ofintracellular states using a novel Covariation Network analysis〇國重 莉奈1、野口 誉之2、米谷 信彦3、中津 大貴1、村田 昌之1,2,4、加納 ふみ1,4
1

東京工業大・科創研・細胞制御工学、2東京大・国際高等研・IRCN、3（株）ニコン、4東京工業大学・マルチモーダル細胞

解析協働研究拠点

To capture the spatiotemporal dynamics of biomolecules for examining the pharmacological action of drugs, wedeveloped a novel protein network analysis, Protein Localization and Modification-based Covariation Network(PLOM-CON) analysis method. This method quantifies the temporal changes in quantity, quality (posttranslational modifications such as phosphorylation), and (co)localization of proteins that are activated orinactivated in response to specific signals input from the immunofluorescence images, and visualizes them as a"covariation network" using the "strength of temporal correlation" of the feature quantities.
In this study, we performed PLOM-CON analysis method to obtain covariation networks for ~50 proteins ininsulin-stimulated rat hepatoma H4IIEC3 cells. The results showed that both Akt, a central molecule in insulin
signaling, and its phosphorylated form p-Akt (Ser473) are the hub of the network. Furthermore, we revealed
that the actin domain, a specific structure temporally created upon insulin stimulus, is the site of accumulationof various molecules for the initiation of glycogen synthesis via phosphorylated GSK3β. Thus, the PLOM-CON
analysis method can visualize how signals are widely propagated into cells, and can be used as a technology thatcan capture the main and side effects of drugs.

Copyright © The Japanese Pharmacological Society2-B-P-108

一般演題（ポスター）

Copyright © The Japanese Pharmacological Society2-B-P-109

一般演題（ポスター）

NMDA-induced activation of the CaMKII-RhoA-Rho-kinase pathway regulates
aversive learning〇船橋 靖広1,2、Ahammad Rijwan Uddin1,2、張 心健3、Emran Hossen1,2、Faruk Md. Omar1,2、王 緩緩4、呉 敏華5、許 伊凡
1,2、坪井 大輔1,2、西岡 朋生1,2、黒田 啓介4、天野 睦紀4、崎村 建司6、内野 茂夫7、山田 清文5、永井 拓3、貝淵 弘三1,2,4

藤田医科大・医科学研究センター・神経・腫瘍のシグナル解析プロジェクト研究部門、2藤田医科大・精神・神経病態解1明センター・細胞生物学部門、3藤田医科大・精神・神経病態解明センター・神経行動薬理学研究部門、4名古屋大・院
医・神経情報薬理学、5名古屋大・院医・医療薬学、6新潟大・脳研究所・モデル動物開発分野、7帝京大・理工・バイオサ
イエンス学科

Glutamate induces Ca2+ influx in neurons through NMDA receptors (NMDARs) and activates Ca2+-dependent
protein kinases, including CaMKII, which play critical roles in synaptic plasticity and learning. However, howthese kinases regulate synaptic plasticity and learning remains largely unknown. Here, we performed
phosphoproteomics and identified 160 proteins including ArhGEF2 whose phosphorylation were promoted by
NMDA. CaMKII phosphorylated ArhGEF2 and stimulated its RhoGEF activity. Aversive stimuli induced
CaMKII-mediated ArhGEF2 phosphorylation and Rho-kinase/ROCK activation in the nucleus
accumbens (NAc). Inhibition of Rho-kinase in the NAc attenuated aversive learning. We also screened Rhokinase substrates and identified 221 proteins including Shank3 which links actin filaments with NMDARs and
AMPA receptors via Dlgap3. The Rho-kinase-mediated phosphorylation of Shank3 increased its interaction with
Dlgap3. Manipulation of Shank3 in the NAc regulated dendritic spine formation and aversive learning in aphosphorylation-dependent manner. These results demonstrate that NMDA activates the CaMKII-ArhGEF2Rho-kinase pathway to induce Shank3 phosphorylation for aversive learning.

Copyright © The Japanese Pharmacological Society2-B-P-110

一般演題（ポスター）

Effects of S-allyl-L-cysteine on phosphorylation of insulin-like growth factor
type-I receptor tyrosine kinase in primary cultures of adult rat hepatocytes.

〇茂木 肇、荻原 政彦、木村 光利
城西大・薬We previously reported that -allyl-L-cysteine (SAC)-induced cell proliferation was involved in intracellular
IGF-I secretion via Janus kinase 2 (JAK2) / phospholipase C (PLC) pathway in primary cultures of adult rat
hepatocytes. In this study, we investigated the involvement of IGF-I receptor tyrosine kinase (RTK) in the SACinduced hepatocyte proliferation by using Western blot analysis. IGF-I RTK (p95 kDa) phosphorylation in
cultured hepatocytes peaked 20 min after SAC stimulation. SAC-induced IGF-I RTK phosphorylation was
suppressed not only by the IGF-I RTK inhibitor AG538 but also by the JAK2 inhibitor TG101209 and the PLC
inhibitor U-73122. Furthermore, the SAC-induced cell proliferation was significantly suppressed by PI3 kinase
inhibitor LY294002, MEK inhibitor PD98059, and mTOR inhibitor rapamycin. These results suggested that the
SAC-induced IGF-I RTK phosphorylation is mediated by JAK2/PLC phosphorylation and subsequently released
IGF-I phosphorylates IGF-I RTK. Then hepatocyte proliferation may be induced via IGF-I RTK / PI3 kinase /MEK / mTOR pathway.

Copyright © The Japanese Pharmacological Society2-B-P-111

一般演題（ポスター）

Cytoprotective effect of Rab proteins against PRAF protein-induced cytotoxicity〇渡部 正彦
帝京大・医療共通教育研究セ

The prenylated Rab acceptor 1 (PRA1) superfamily member PRAF3 plays crucial roles in membrane traffic as a
GDI displacement factor

protein interaction with a variety of Rab proteins, as well as in the modulation ofantioxidant glutathione through its interaction with the amino acid transporter EAAC1. It is known that theoverexpression of PRAF3 induces the toxicity of the host cell, however, the factors capable of cancelling thecytotoxicity remained unknown. Our findings demonstrate that Rab1a can protect from the cytotoxicity of
PRAF3-overexpressed human cells. Cytoprotective effects of Rab1a protein could further suggest that PRAF3
and Rab1a are closely related to each other physiologically and genetically.

Copyright © The Japanese Pharmacological Society2-B-P-112

一般演題（ポスター）

Roles of MFN2 and MFN2 associated protein in chemotaxis of neutrophil-like
differentiated HL-60 cells〇真崎 雄一1、東 恒仁1、小林 純子2、小野寺 康仁31

北海道大・院医・細胞薬理、2北海道大・院医・組織細胞、3北海道大・医・医理工

Neutrophils are important in innate immunity and in the initiation of an acute response to infection. Under
normal conditions, the mitochondrial membrane potential of neutrophils is low, and neutrophils energy depends
fundamentally on glycolysis. In contrast, neutrophils require energy supplied from mitochondrial oxidative
phosphorylation (OXPHOS) during infection. The inhibition of mitochondrial OXPHOS blocks the chemotaxisof neutrophils. Here, we examined the mitochondrial morphology of neutrophil-like differentiated HL-60 cells
after chemoattractant-formyl-Met-Leu-Phe ( MLP) stimulation. We found that mitochondrial morphology

changes to a tubular form after MLP stimulation. Mitochondrial OXPHOS activity and mitochondrial complex
II significantly increased after MLP stimulation. On the other hand, the silencing of mitochondrial fusion
protein() suppresses mitochondrial morphological changes.

silencing suppressedOXPHOS activation and chemotaxis after MLP stimulation. Furthermore, the silencing of MFN2 associatedprotein suppresses also mitochondrial morphological changes and chemotaxis upon MLP stimulation. These
results suggest that MFN2 and MFN2 associated protein are involved in chemotaxis of differentiated HL-60
cells.

Copyright © The Japanese Pharmacological Society2-B-P-113

一般演題（ポスター）

Cigarette smoke gas phase induces ferroptosis via PKC in tracheal epithelialcells〇東 恒仁1、眞井 洋輔2、真崎 雄一1
1

北海道大・院医・細胞薬理、2北海道大・院医・皮膚科

Cigarette smoking is one of the risk factors for respiratory diseases such as chronic obstructive pulmonary
disease and emphysema. Cigarette smoke can be divided into two phases; tar (particle) phase including nicotine
and gas phase. Although both phases of cigarette smoke have cytotoxic activity and affect respiratory system, themolecular mechanism for cytotoxicity has remained to be clarified. In this study, we have examined the effects ofcigarette smoke extracts on tracheal epithelial cells and lung cells. Both tar and gas phases induced cell death.
Ferrostatin-1 suppressed gas phase-induced cell death, whereas it had no effects on tar phase-induced cell
death. Several unsaturated carbonyl compounds such as acrolein (ACR) and methyl vinyl ketone (MVK) aremajor cytotoxic compounds in the gas phase. Ferrostatin-1 also suppressed ACR- and MVK-induced cell deathin tracheal epithelial cells. These results indicate that ACR and MVK in gas phase are critical factors forferroptosis induction by cigarette smoke in the respiratory system. Since we have previously reported that ACRand MVK-induced cell death is PKC-dependent in aorta smooth muscle cells, we have examined PKC inhibitors.
The results suggest that novel and/or atypical PKCs involve in cigarette smoke-induced ferroptosis induction intracheal epithelial cells.

Copyright © The Japanese Pharmacological Society2-B-P-114

一般演題（ポスター）

Involvement of glycerophosphodiesterase 7 in the intracellular production of
cyclic phosphatidic acid〇北風 圭介1、Ali Hanif2、木本 来希1,3、竹之内 康広1、石丸 浩靖1、山下 純4、上田 夏生5、田中 保2、岡本 安雄1、坪
井 一人1
川崎医大・医、2徳島大・院生物資源、3奈良医大・医、4帝京大・薬・生物化学、5香川大・医・生化学

1

Cyclic phosphatidic acid (cPA) is a lipid mediator present in tissues and plasma, which regulates physiologicaland pathological processes via the suppression of peroxisome proliferator-activated receptor γ (PPARγ).
Glycerophosphodiesterase 7 (GDE7) is an endoplasmic reticulum-localized lysophospholipase D-type enzyme,
and mouse GDE7 was reported to catalyze the production of cPA in cell-free systems. However, it remains
unknown whether this reaction occurs in living cells. In this study, we found that overexpression of human
GDE7 in COS-7 cells increased not only cPA-producing activity in the membrane fraction but also cPA levels in
living cells. Furthermore, we demonstrated that active site of human GDE7 is directed toward the luminal sideof the endoplasmic reticulum membrane. Mutagenesis revealed that amino acid residues F227 and Y238 ofhuman GDE7 play an important role in cPA production. Finally, GDE7-knockout decreased the cPA levels andderepressed mRNA expression of PPARγ target genes in human mammary MCF-7 cells, suggesting that cPA
acts as an intracellular lipid mediator. These findings lead to a better understanding of the biological role of
GDE7 and cPA in cells.

Copyright © The Japanese Pharmacological Society2-B-P-115

一般演題（ポスター）

Establishment of the novel method to culture primary neurons from aged rodent
brain〇笠井 悠哉1、野崎 千尋2、柴田 重信1
早稲田大・先進理工学部・電気・情報生命工学科、2早稲田大・国際理工学センター・Major in Bioscience

1

Recent study showed that cannabinoid CB1 receptor-mediated signaling may take a huge role in age-related
modification of brain function. CB1 deficient animals show significant cognitive deficit in old age, however theyshow better learning ability in young age. Even though the finding is surprising, the molecular mechanisms ofsuch difference, especially for better learning in young age remains unknown mainly due to lack of suitable invivo/ex-vivo models. Here we will purpose to use the primary cultured neurons as the ex-vivo model of aging,which can be enabled with gentleMACS technology. This method is said to enable the primary cell culture fromadult rodents but up to age of P60. In the present study we aimed to establish a method for primary culture ofbrain neurons using aged mice, which can be also used for knockout animals of CB1 receptors.
We first tried to prepare the primary culture of brain neurons from neonatal (P7), young (P50) and mature(P105) mice. After 3-days of culture we could find that all of them survived well and start to extend their axons.
Notably, P105 is twice an older age than previously reported suggesting that this method could be usable toevaluate the neuronal growth and cellular activity after the certain aging. Further, they could even survive fornext 10-days with continuous axon growth. Using this method, we will further conduct the primary cell culture
in knockout animals of CB1 as well as endocannabinoid producing enzyme DAGL-a to see the effect of
endocannabinoid deficiency to the neuronal growth and cellular activity.

Copyright © The Japanese Pharmacological Society2-B-P-116

一般演題（ポスター）

Differences in expression of fat metabolism-related genes in liver and adipose
tissue after ingestion of high-fat diet in obese and obesity-resistant mousestrains〇山口 瑞希1、神原 遥1、渕 美樹1、吉田 真尋1、金子 啓一郎1、藤田 融1、前田 利男2、田邉 由幸1
横浜薬科大・薬・薬理学研究室、2静岡県立大・薬・臨床薬剤学講座

1

We have isolated inbred mouse strains that stably express either obesity-prone (ddY-H) or obesity-resistant(ddY-L) phenotypes. The ddY-H mice spontaneously develop hyperglycemia and hepatic steatosis along with asignificant increase in body weight and fat mass even by fed with normal diet, whereas the ddY-L mice hardly
develop these metabolic syndrome-like phenotypes. To investigate metabolic differences between ddY-H andddY-L mice during high-fat diet (HFD) feeding, we analyzed the expression of transcripts related to glucoseand fat-metabolisms in the liver and adipose tissues. At 6 weeks of age with normal diet, no significant
difference was observed in the expression of CD36, a major fatty acid transporter, in the liver of both strains,
although the expression of PPARγ, a key transcription factor for fat synthesis, tended to be higher in the liverof ddY-H mice than that of ddY-L mice. Afterward, ingestion of HFD from 6 to 9 and 12 weeks of age
dramatically induced the expression of PPARγ and CD36 in the liver of ddY-H mice. Neither PPARγ nor
CD36 was induced in the liver of HFD-fed ddY-L mice, however, both transcripts were induced significantly in
epididymal fat tissue. These results suggest that the low efficiency of hepatic expression of PPARγ and CD36
may be involved in the obesity-resistant phenotype of ddY-L mice.

Copyright © The Japanese Pharmacological Society2-B-P-117

一般演題（ポスター）

Involvement of glucose on TGF-β1-induced epithelial-mesenchymal transition inepithelial keratinocytes〇武石 幸容1、長岡 良礼1、髙橋 千代1,2、武田 佳奈1,2、岡村 和彦3、大徳 浩照4、八田 光世11

福岡歯科大・歯・細胞分子生物学講座 分子機能制御学分野、2福岡歯科大・矯正歯科学分野、3福岡歯科大・歯・病

態構造学分野、4筑波大・生存ダイナミクス研究センター

Epithelial-to-mesenchymal transition (EMT) is a unique program in which epithelial cells become more likemesenchymal cells. EMT is involved in several processes, including development, wound healing, fibrosis, andcancer progression, but the molecular mechanism remains to be elucidated.
We analyzed the involvement of glucose on transforming growth factor (TGF)-β1-induced EMT in humankeratinocyte HaCaT cells. Using fluorescent staining, TGF-β1-treated HaCaT showed the formation of actin
stress fiber, regardless of glucose concentration. EMT-related markers were partially suppressed in TGF-β1treated cells under low-glucose conditions. Furthermore, both intracellular glucose and lactate, a glucose
metabolite, were decreased under low-glucose conditions. We focused on the relationship between autophagy
and the partial suppression of EMT under low-glucose conditions because we found in a previous study that LC-3 and GABARAPL1, autophagy markers, may be associated with TGF-β1-induced EMT. The expressionof those markers were regulated in a glucose- and TGF-β1-dependent manner.
Therefore, it is likely that TGF-β1-induced EMT is regulated by glucose-induced autophagy in epithelial
keratinocytes.

Copyright © The Japanese Pharmacological Society2-B-P-118

一般演題（ポスター）

Investigation of dephosphorylation of synaptic phosphoproteins engaged in
mammalian sleep-wake regulation〇曹 思鈺1、戸根 大輔1、山田 陸裕2、隅山 健太3、上田 泰己1,2
東京大・院医、2理研・生命システム研究センター・合成生物学チーム、3理研・生命システム研究センター・高速ゲノム

1

変異マウス作製研究チーム

Sleep-wake cycle is an organism-level phenomenon that is precisely controlled by multi-layered systems such as
circuits, cellular and molecular levels in a brain. Recent studies have provided a deeper understanding of themolecular basis of mammalian sleep-wake regulation and some studies suggested dynamic changes in neuronalprotein phosphorylation under the control of the sleep-wake cycle. However, the core molecular mechanism of
the dynamic changes in phospho-proteins and whether it could drive the transition between sleep and wake is
still unclear. In this study, we identified a gene known to be involved in the dephosphorylation process in various
signaling pathways in mammals as a novel sleep-regulating factor. Exogenous expression of the active form of
GeneX in excitatory postsynapses resulted in a significant increase in sleep duration. Besides, knockout of one ofgeneX regulators which works as the scaffold protein of the proteinX in excitatory postsynapses resulted in asignificant decrease in sleep duration. These results suggest that the sleep-wake cycle is modulated bydephosphorylation processes involving proteinX in the excitatory postsynapses.

Copyright © The Japanese Pharmacological Society2-B-P-119

一般演題（ポスター）

Neuronal nitric oxide synthase is regulated by supersulfides〇土屋 幸弘、荒木 笙馬、渡邊 泰男
昭和薬科大・薬・薬理

Neuronal nitric oxide synthase (nNOS) is a Ca2+/calmodulin (CaM)-dependent enzyme thatcatalyzes the metabolism of L-arginine (Arg) to nitric oxide (NO) in the presence of NADPH and O2. nNOS isonly active as a dimer, however, a reduction of cofactor tetrahydrobiopterin (BH4) levels drastically destabilizes
the dimer and thereby uncoupling from NADPH and production of superoxide rather than NO occur. Protein
polysulfidation at specific Cys residues plays pivotal roles in the protein function. In this study, we investigatedthe molecular mechanism of supersulfides-induced inactivation of nNOS.
Incubation of recombinant nNOS with Na2S4 in the presence of Arg and BH4 resulted
in a dose-dependent inhibition of NO production which was cancelled by reducing agent, dithiothreitol.
Likewise, NADPH oxidation was inhibited with Na2S2 in the presence of BH4 either with or without Arg but was
not inhibited when both Arg and BH4 were omitted. This indicates that inhibition targets NADPH oxidation
with dimer formation of nNOS and without sign of uncoupling. The dimer/ monomer ratio of nNOS was notaffected by Na2S4. Treatment of Na2S4 resulted in decreased ADP- but not CaM-agarose binding ability tonNOS. We successfully did generate the Na2S4-insensitive nNOS mutant in that its specific Cys reside within the
NADPH-binding domain were mutated with serine. Thus, NADPH oxidation activity of dimerized nNOS isregulated by supersulfides via its regulatory cysteine residue modification.

Copyright © The Japanese Pharmacological Society2-B-P-120

一般演題（ポスター）

Search for tyrosine kinases overexpressed in the hypertrophied right ventricularwall of monochromotaline-induced pulmonary hypertensive rats

〇田邉 由幸、金子 啓一郎、吉田 真尋、藤田 融
横浜薬科大・薬Pulmonary arterial hypertension (PAH) results in right ventricular failure due to markedly elevated pulmonary
arterial pressure and compensatory right ventricular hypertrophy. Although the involvement of tyrosine kinase(PTK) in the smooth muscle pathogenicities of PAH has been suggested, attempts to treat with a “multi-target”
PTK inhibitor (PTKI) have not made sufficient progress due to harmful effects such as cardiotoxicity. In thisstudy, we searched for PTK, whose expression is selectively increased in the hypertrophied right ventricular wallof PAH rats. RT-PCR targeting PTK domain consensus sequence was carried out for right ventricular wall (RV)
and left ventricular wall with septum (LV+S) from monocrotaline-induced (MCT-) PAH rats and control rats.
Southern hybridization was performed using various PTK probes derived from MCT-PAH rat RV. Selective
increased expression of at least three types of PTK in MCT-PAH rat RV was observed, including PDGF-Rβ,
which we previously reported its overexpression in pulmonary arteries of MCT-PAH rats. There was nodifference in the expression of c-abl, which is thought to be involved in PTKI cardiotoxicity, in the LV+S and
RV in both PAH and control rats. Our results suggest that a more specific inhibition of PTK isoform would besuitable for safely treating PAH using PTKI.

Copyright © The Japanese Pharmacological Society2-B-P-121

一般演題（ポスター）

The production of hydrogen sulfide in glioblastoma cell lines〇澁谷 典広、佐藤 彩湖、河津 咲穂、伊藤 凌大、木村 英雄
山口東京理科大・薬Hydrogen sulfide (H2S) functions as a signaling molecule and a cytoprotectant. The pathway to produce H2S
includes cystathionine β-synthase (CBS), cystathionine γ-lyase (CSE), and 3-mercaptopyruvate
sulfurtransferase (3MST). 3MST produces H2S from 3-mercaptopyruvate that is provided from l-cysteine andα-ketoglutarate (α-KG) by cysteine aminotransferase (CAT) and from d-cysteine by d-amino acid oxidase(DAO). Recently, a new concept emerges in H2S biology, showing tumor cells upregulate the production of H2S
and utilize it to promote tumor growth and chemotherapy resistance. In this study, we investigated the
production of H2S in cell lines derived from glioblastoma, the most common- and aggressive-type of brain
tumors among adults. Lysates of glioblastoma cells produced H2S from l-cysteine and α-KG. In the absence of
α-KG, the production of H2S was significantly decreased, suggesting that H2S production is dependent on theactivity of CAT. The production of H2S from l-cysteine and α-KG was correlated with the amount of boundsulfane sulfur, a storage form of H2S. Unlike the l-cysteine pathway, d-cysteine pathway does not operate in the
glioblastoma cells. Further investigation will be needed to clarify whether regulation of the l-cysteine pathwaychanges proliferation and anti-cancer drug resistance of the glioblastoma cells.

Copyright © The Japanese Pharmacological Society2-B-P-122

一般演題（ポスター）

Behavioral analysis of mice overexpressing PP5 specifically in the mice CA3
region of hippocampus.

〇宇野 恭介、河原井 康介、金城 俊彦、倉本 展行
摂南大・薬[Purpose] We have explored the possibility that protein phosphatase 5 (PP5), together with PP1 and PP2A,
promotes desensitization of GABAB receptors and attenuates receptor action by dephosphorylating the 892nd
serine residue of mouse GABAB receptors. In this study, we investigated the possibility that dephosphorylationof GABAB receptors in the hippocampal CA3 region affects the behavior of mice.
[Methods]The PP5 gene was cloned in a pAAV-CMV-GFP vector to create an adeno-associated viral vector(AAV). Mice in which the PP5 gene was overexpressed in the hippocampal CA3 region (PP5 mice) using thecreated AAV or mice in which AAV containing only the GFP gene was introduced as a control group (GFP
mice) were created, and the effect on mouse behavior was analyzed.
[Results] In the Open field test, PP5 mice tended to stay in the center area longer than GFP mice, and asignificant difference was observed especially for last 10 minutes of the session. In an elevated cross-maze test,
PP5 mice tended to stay longer in open zones than GFP mice. In the Y-maze test, PP5 mice showed adecreasing trend in alternation behavior compared to GFP mice. In the novel object search test, a significant
increase in the time of interest in the novel object was observed in GFP mice, but in PP5 mice there was no
significant difference in the time of interest in the two objects.
[Conclusion] The behavioral experiments show that overexpression of PP5 in the CA3 region may affect mice toreduce anxiety-like behavior and cognitive function.

Copyright © The Japanese Pharmacological Society2-B-P-123

一般演題（ポスター）

A novel endothelin A receptor antagonist eliminates the analgesic tolerance andrecovers the attenuated analgesic effects for long-term use of opioids〇大島 佳織1,2、野中 美希2、黒田 唯2,3、宮野 加奈子2、高柳 広1、上園 保仁2
東京大・院医・病因・病理学、2東京慈恵会医科大・医・疼痛制御、3順天堂大・医・麻酔科学・ペインクリニック

1

Endothelin-1 (ET-1) is known as a vasoconstrictor. Several studies have reported that ET-1 induced pain signals
through the ETA receptor (ETAR). Although ETAR antagonists are reported to enhance analgesic effects of
opioid agonists and relieve its tolerance, the mechanisms remain unclear. We previously revealed the novel andhighly selective ETAR antagonist (provided by Eisai Co., Ltd.) enhanced morphine-induced opioid receptor(OR) activities with HEK293 cells stably expressing either ETAR (ETAR cells), μOR (μOR cells), or both
ETAR and μOR (ETAR/μOR cells). Further, we found that μOR and ETAR form a dimer as a possible targetof this antagonist, and ET-1 internalized the ETAR/μOR heterodimer into the cytosol, resulting in decreased
numbers of μOR on the cell membrane, followed by reduced responses by morphine. There are opioid agonists
prone to internalize μOR or not; morphine is hardly to internalize, whreas fentanyl preferentially internalizes
μOR among opioid agonists. Such difference in μOR internalization is believed to involve in the analgesic
tolerance. In the present study with cells expressing ETAR/μOR, we evaluated the effects of the ETAR
antagonist on the enhancement of μOR activities by several opioid agonists in addition to morphine, andcompared the selectivity of the ETAR antagonist as an analgesic adjuvant.

Copyright © The Japanese Pharmacological Society2-B-P-124

一般演題（ポスター）

Role of arachidonic acid-containing phospholipids in neuropathic pain

〇山本 将大1、清水 孝雄1,2、進藤 英雄1,3
1(国研)国立国際医療研究センター研究所・脂質生命科学研究部、2公益財団法人微生物化学研究会 微生物化学研究所、3東京大・院医・脂質医科学連携講座

Glycerophospholipids (PLs) play pivotal roles in our body as main structural components of biologicalmembranes and precursors of lipid mediators. Recently, we revealed that peripheral nerve injury (PNI)
increased arachidonic acid-containing PLs (ARA-PLs) in the dorsal root ganglion (DRG), spinal microglia, andspinal astrocytes. Therefore, we focused on LPCAT3 (also called LPLAT12), a biosynthetic enzyme of ARAPLs, and established cell type-specific(

CreERT2), and satellite glia/astrocyte (Cre-knockout mice: DRG neuron (), macrophage/microglia-Cre). We found that PNI-induced mechanical allodynia wereattenuated in DRG neuron-specific and satellite glia/astrocyte-specific-knockout mice. These results

suggest that alteration of quality of lipid in these cells may be a cause of neuropathic pain. Now, we are
analyzing the mechanisms by which ARA-PLs are involved in neuropathic pain.

Copyright © The Japanese Pharmacological Society2-B-P-125

一般演題（ポスター）

Synergistic effect of topical application of TRP channel antagonists to gingiva on
orthodontic force-induced pain in rats

〇湯川 未郷1、佐藤 慶太郎2、須田 直人1、安達 一典2
明海大・歯・形態機能成育学講座 歯科矯正学分野、2明海大・歯・病態診断治療学講座 薬理学分野

1

The electrical stimulation to gingiva induced the jaw-opening reflex (JOR). We have previously reported thatthe threshold for inducing jaw-opening reflex (JOR-TH) is significantly reduced by application of orthodontic
force to teeth, and the intraperitoneal administration of TRPV1 antagonist recovers the JOR-TH. However,serious adverse side effects (e.g., hyperthermia) are reported by general administration of TRPV1 antagonists,and effective and safety application has to be established. In this study, TRPV1 and or TRPA1 antagonists weretopically applied to gingiva, and the orthodontic force-induced JOR excitation was investigated with otherfeatures (e.g., trigeminal excitation, inflammatory cytokine alteration, rectal and gingival temperatures, andtemperature sensitivity of the plantar and gingiva). Topical application of TRP antagonists immediately (D0)or one-day (D1)after orthodontic force application significantly increased JOR-TH the next day in dosedependent and synergistic manner. On the other hand, there were no alterations in rectal and gingival
temperature. In D0, the analgesic effect was associated with a significant reduction of CINC2 (cytokine-induced
neutrophil chemoattractant-2) in the periodontium. In addition, chemical application-induced increase intemperature sensitivity was only observed in plantar. Taken together, topical application of TRP antagonists
cocktail may reduce clinical orthodontic pain via CINC-2 reduction without adverse side effects.

Copyright © The Japanese Pharmacological Society2-B-P-126

一般演題（ポスター）

PAC1 receptor antagonist PA-81004 provides an excellent preventive effectagainst oxaliplatin-induced cold allodynia〇足原 佑弥1、斎藤 弘樹2、宮田 篤郎2、栗原 崇2、髙﨑 一朗1
富山大・院医薬理工・生体情報薬理、2鹿児島大・院医歯・生体情報薬理1

Copyright © The Japanese Pharmacological Society2-B-P-127

一般演題（ポスター）

Effects of meloxicam and morphine on nociceptive behaviour in rats withbilateral intraplantar injections of carrageenan into hind paws

〇三枝 禎、川島 央暉、青野 悠里
日本大・松戸歯・薬理Intraplantar injection of carrageenan into the hind paw of rodents is used to induce experimental inflammatory
symptoms, including nociceptive behaviour. The unilateral injection of carrageenan into a hind paw to provoke
inflammation was reported to enhance nociceptive sensitivity in the contralateral paw in rats. In order to provide
balanced stimulation to the both sides, carrageenan should be injected bilaterally into the hind paws. In thepresent study we analysed the effects of systemic administration of meloxicam, a non-steroidal antiinflammatory drug, or morphine on paw withdrawal and threshold forces in response to tactile stimuli in rats
with bilateral intraplantar injections of carrageenan into hind paws. For comparison, effects of these analgesic
drugs were also analysed in rats with a bilateral sciatic nerve ligation to produce neuropathic pain. In bothcarrageenan-treated and sciatic nerve-ligated rats, the paw withdrawal responses and threshold forces in
response to tactile stimuli were increased and decreased, respectively. These nociceptive behaviours ofcarrageenan-treated rats were each inhibited by meloxicam and morphine. Morphine, but not meloxicam,
inhibited the behavioural changes in sciatic nerve-ligated rats. The inhibitory effect of morphine in carrageenantreated rats was much larger than that in nerve-ligated rats. These results suggest that rats with bilateral
intraplantar injection of carrageenan into hind paws can be considered an animal model of inflammatory pain.

Copyright © The Japanese Pharmacological Society2-B-P-128

一般演題（ポスター）

Pain-induced potentiation at synapses between pain-activated neurons in theparabrachial nucleus and central amygdala of FosTRAP mice

〇内山 瑳和子1,2、奥田 崇雄2、高橋 由香里2、津田 誠1、加藤 総夫2
九州大・院薬・薬理学分野、2東京慈恵会医科大・医・神経科学研究部

1

The lateral parabrachial nucleus (LPB) collects nociceptive information from the spinal cord and trigeminal
sensory system and sends it to the central nucleus of the amygdala (CeA). This LPB-CeA pathway undergoes
extensive synaptic plasticity in various rodent models of persistent pain and plays an essential role in“nociplastic” expression of widespread sensitization (Miyazawa et al, 2018; Sugimoto et al, 2021). However, as
this pathway also transmits signals other than nociception, it remains elusive whether the plastic changes at
LPB-CeA synapses are specific to pain-associated neuron pairs. We selectively activated synaptic transmission
between pain-associated pairs of LPB and CeA neurons using the FosTRAP technique, in whichchannelrhodopsin-2 (ChR2) and td-Tomato were expressed in LPB neurons and CeA neurons, respectively,
following persistent inflammatory pain to activate c-Fos expression in these regions. In brain slices from the
FosTRAP mice, we recorded light-evoked excitatory postsynaptic currents (leEPSCs) from “FosTRAPed” CeA
neurons and found 1) leEPSC amplitude was larger specifically in these pairs and 2) a second-shot stimulation
given 4-8 weeks after the first one resulted in larger leEPSC amplitude in these pairs, which was, unexpectedly,
observed in animals also with non-inflammatory brief stimulation. These results suggest that the LPB-CeA
synaptic potentiation occurs specifically between pain-associated pairs, which would be primed for plasticchanges in response to future aversive events of various types.

Copyright © The Japanese Pharmacological Society2-B-P-129

一般演題（ポスター）

Role of spinal cord-projecting cortical neurons in Aβ fiber-derived neuropathic
allodynia in mice

〇藤森 一樹、津田 誠
九州大・院薬・薬理学分野

Mechanical allodynia, pain caused by innocuous mechanical stimulation, is a characteristic symptom ofneuropathic pain that develops after peripheral nerve injury (PNI). In this study, we used Thy1-ChR2 mice, atransgenic mouse line that expresses channelrhodopsin-2 in touch-sensing myelinated primary afferent fibers
but not in nociceptors. Light illumination to the plantar skin of Thy1-ChR2 mice with PNI produced pain-likewithdrawal behavior and increased c-FOS expression in superficial spinal dorsal horn (SDH) neurons. This
light-induced pain-like behavior disappeared by silencing Aβ fibers, but was not suppressed by morphine
administration. Furthermore, using pathway-selective gene expression by adeno-associated viral vectors, wedemonstrated that chemogenetic silencing of primary sensory cortex (S1) neurons projecting to the SDH
attenuated pain-like behavior and reduced the number of c-FOS-expressing SDH neurons evoked byphotostimulation of Aβ fibers. These findings indicate that spinally projecting S1 neurons contribute to Aβ
fiber-derived neuropathic allodynia.

Copyright © The Japanese Pharmacological Society2-B-P-332

一般演題（ポスター）

Augmented mechanical response of spinal dorsal horn neurons in a rat model offibromyalgia.

〇歌 大介1、坪島 功幸2、西条 寿2、水村 和枝3、田口 徹41

富山大・院医薬・応用薬理、2富山大・院医薬・システム情動、3日本大・歯・生理学、4新潟医療福祉大・リハビリテーショ

ン学部

Purpose: Fibromyalgia (FM) is characterized by chronic widespread pain with mechanical allodynia andhyperalgesia. However, the neural mechanisms of nociception/pain are poorly understood. The aim of this study
was to examine the responsiveness of superficial dorsal horn (SDH) neurons using a rat model of FM.
Methods: Reserpine, a depleter of biogenic amines in the nervous system, was subcutaneously injected to makethe mode. Extracellular recording of the SDH neurons

was performed at the spinal segments L4/L5

under urethane anesthesia. Mechanical stimulation with a series of calibrated von Frey filaments was applied for
10 seconds to an identified receptive field of the SDH neurons.
Results: The SDH neurons showed mechanical stimulus intensity-dependent increases in the discharge rate
both in the control and the reserpine-injected group. However, the magnitude of the mechanical response wassignificantly greater in the reserpine-injected group. Some SDH neurons in the reserpinized rats exhibitedbackground discharges of low frequencies, although those in the control rats did not.
Conclusion: These results suggest that facilitated mechanical sensitivity of the SDH neurons is involved inmechanical allodynia and hyperalgesia in a rat model of reserpine-induced pain. Similar spinal mechanisms may
underlie in FM patients.

Copyright © The Japanese Pharmacological Society2-B-P-130

一般演題（ポスター）

Effects of calcium channel blockers on stiffness of the aortic and femoral arterial
segments in anesthetized rabbits: comparison of the actions of nifedipine andcilnidipine〇佐藤 啓1,2、鈴木 保菜実1、佐久間 清1,2、千葉 達夫1,3、相本 恵美1、永澤 悦伸1、高原 章1
1

東邦大・薬・薬物治療、2東邦大医セ佐倉病院・薬剤部、3東邦大医セ大橋病院・薬剤部

Ca2+ channel blocker is one of the vasodilators with excellent efficacy in preserving organ blood flow, which may
be partly affected by functional stiffness of the conduit arteries. In this study, we assessed effects of L-type Ca2+
channel blocker nifedipine and L/N-type Ca2+ channel blocker cilnidipine on stiffness of the aortic and femoral
arterial segments (aortic β and femoral β, respectively) in anesthetized rabbits. The methodology to obtainaortic β and femoral β was essentially same as that for cardio-ankle vascular index with the VaSera devise forhuman. Antihypertensive dose of nifedipine (300 µg/kg, i.v.) increased the aortic β but hardly affected thefemoral β. However, nifedipine decreased the femoral β in the presence of an α-adrenoceptor blocker
doxazosin (1 mg/kg, i.v.). Antihypertensive dose of cilnidipine (30 µg/kg, i.v.) increased the aortic β but
decreased the femoral β. These results suggest that the increment of the aortic β reflecting the stiffening of
the aortic segment and the decrement of the femoral β reflecting the softening of the femoral arterial segment
are common effects of Ca2+ channel blockers on the conduit arteries. In addition, the stiffness of femoral arterial
segment is considered to be modified by vasoconstriction associated with sympathetic reflex.

Copyright © The Japanese Pharmacological Society2-B-P-131

一般演題（ポスター）

Possible pathophysiological importance of C-terminal tyrosine phosphorylationof CaV1.2 in vessel remodeling〇冨田 拓郎1、川岸 裕幸1、中田 勉2、山田 充彦1
1

信州大、2信州大・基盤研究支援センター・機器分析部門

Voltage-dependent CaV1.2 L-type calcium channels play a critical role in the regulation blood pressure. Besides,
CaV1.2 is implicated to play a pivotal role in chronic vessel remodeling such as atherosclerosis because several
clinical trials indicate that CaV1.2 inhibitors delay the progression of atherosclerosis. However, how CaV1.2
participates in atherogenesis still remains obscure. Vascular smooth muscle cells (VSMC) are critically involvedin atherogenesis. Medial VSMCs migrate and proliferate into the intima and form a part of plaque uponendothelial injury. Recently, we have identified that two tyrosine residues in the C-terminus (Tyr1709 andTyr1758) of VSMC CaV1.2 are phosphorylated by Src-family kinases in response to PDGF.

studiesdemonstrated that PDGF enhances CaV1.2 channel activity through this phosphorylation, thereby induceing
VSMC migration. Thus, CaV1.2 may contribute to vessel remodeling in atherosclerosis. In order to verify thishypothesis, we established a knock-in mice line harboring a mutation at one of the tyrosine residues (Y1709F byusing the CRISPR/Cas9 system. Although these mice exhibited no obvious developmental defects, we are now
analyzing the effect of carotid artery ligation, a model of intimal thickening in these mice.

Copyright © The Japanese Pharmacological Society2-B-P-132

一般演題（ポスター）+

Differential effects of Na channel blockers on the conduction velocity and theeffective refractory period in the guinea pig left atrium and pulmonary vein
myocardium〇濵口 正悟、土屋 真優、冨山 陽、行方 衣由紀、田中 光
東邦大・薬・薬物

We compared the effects of Na+ channel blockers on the electrophysiological properties in the isolated
pulmonary vein-left atrium connected preparation from guinea pig. Four concentric electrodes were attached tothe pulmonary vein and left atrium to measure the intra-pulmonary vein and intra-atrial conduction velocity andthe effective refractory period. Pilsicainide (10 µM), a Na+ channel blocker, decreased the conduction velocity
and prolonged the effective refractory period in both regions. GS-458967 (1 µM) and eleclazine (10 µM),
known as Na+ channel blockers at open and inactivated states, prolonged the effective refractory period in thepulmonary vein but not in the left atrium without affecting the conduction velocity in both regions. The
wavelength, calculated by the conduction velocity × the effective refractory period, in the pulmonary vein was
prolonged by GS-458967 and eleclazine but shortened by pilsicainide, while these drugs did not prolong that in
the left atrium. These results suggest that certain types of Na+ channel blockers, such as GS-458967 andeleclazine, appear to be effective therapeutic agents for atrial fibrillation in which the reentrant excitation in thepulmonary vein is important.

Copyright © The Japanese Pharmacological Society2-B-P-133

一般演題（ポスター）

Roles of vascular smooth muscle NCX1/2 in the development of pulmonary
arterial hypertension〇根本 隆行1、小松 知広1,2、喜多 知1、田頭 秀章1、上原 吉就2、喜多 紗斗美3、岩本 隆宏11

福岡大・医・薬理学、2福岡大・スポーツ科学、3徳島文理大・薬・薬理学

Pulmonary arterial hypertension (PAH) is a severe and progressive disease that causes right heart failure.
Recent studies suggested that the hypercontraction and excessive proliferation of the pulmonary artery induced
by Ca2+ signaling abnormality may be involved in the pathogenesis of PAH, though their molecular mechanisms
remain unclear. The Na+/Ca2+ exchanger (NCX) is a bidirectional transporter that is controlled by membranepotential and transmembrane gradients of Na+ and Ca2+. Vascular smooth muscle (VSM) NCX plays animportant role in intracellular Ca2+ homeostasis and Ca2+ signaling. In this study, we investigated thepathophysiological roles of NCX1/2 in hypoxia-induced PAH, using VSM-specific NCX1/2 knockout mice and
NCX1/2 inhibitors. VSM-specific NCX1-knockout mice exhibited attenuation of hypoxia-induced PAH andright ventricular hypertrophy compared with wild-type mice. In addition, administration of NCX1 inhibitors
suppressed hypoxia-induced PAH and pulmonary vessel muscularization. These findings indicate that genetic
knockout and pharmacologic inhibition of NCX1 attenuate the development of hypoxia-induced PAH.
Furthermore, our next studies with VSM-specific NCX2-knockout mice will provide new evidence that NCX2
differentially contributes to the development of hypoxia-induced PAH.

Copyright © The Japanese Pharmacological Society2-B-P-134

一般演題（ポスター）

The alteration of contractile reactivity in isolated mesenteric arteries from Zucker
fatty diabetes mellitus rats

船田 拓志、〇大谷 紘資、兒玉 朋子、岡田 宗善、山脇 英之
北里大・獣医・獣医薬理Obesity and type 2 diabetes (T2D) are major risk factors for cardiovascular diseases. Zucker fatty diabetes
mellitus (ZFDM)-

fa/fa(Homo) rats develop obesity and T2D at a young age, while ZFDM-

fa/+(Hetero)rats are normal. We recently reported that blood pressure in Homo is normal until 35 weeks of age, while blood
adrenaline level and sympathetic nervous activity are lower than Hetero. In the present study, we tested the
hypothesis that contractile reactivity to adrenaline in peripheral blood vessels from Homo increases. After theisolated mesenteric arteries were cumulatively administrated with adrenaline (1 nM-30 μM) in the presence orabsence of propranolol (1 μM), isometric tension was measured by a Magnus method. Body weight and blood
glucose levels in Homo were significantly higher than Hetero, while heart rate was significantly lower. The
adrenaline-induced contraction in the isolated mesenteric artery from Homo was significantly higher than
Hetero. In Hetero, pretreatment with propranolol enhanced the adrenaline-induced contraction, while it had noeffect in Homo. These results suggest that the adrenaline-induced contraction in the isolated mesenteric arteryfrom Homo may be increased due to the altered expression of adrenaline receptors. In addition, the increased
adrenaline responsiveness of peripheral arteries in Homo would maintain the blood pressure normal despite thedecrease of sympathetic nerve activity.

Copyright © The Japanese Pharmacological Society2-B-P-135

一般演題（ポスター）

A study on a pathophysiological role of a macromolecular complex of the cardiac
KCNQ1 channel〇黒川 洵子1、服部 希海1、野間口 財1、杉本 早穂1、岩鶴 果奈1、児玉 昌美2、渡邉 泰秀1、清水 聡史1,3、永森 收志3、
坂本 多穗11

静岡県立大・薬・生体情報分子解析学、2順天堂大・医・薬理学教室、3東京慈恵会医科大・医・臨床検査医学

The IKs channels, which contribute to the repolarization phase of the cardiac action potential, are composed of
the alpha subunit KCNQ1 and beta subunit KCNE1. Mutations in these genes are associated with thedevelopment of lethal arrhythmias due to congenital QT prolongation syndrome and are influenced bysympathetic nerve stimulation and sex hormones. As a molecular mechanism, we have demonstrated the
involvement of a molecular complex of the KCNQ1 channel in the Ks regulation by intracellular Ca2+, cAMP, and
NO. Recently, membrane proteomics has shown an association between the KCNQ1 molecular complex and Ca2+

signaling, but the pathophysiological role of this association has not been elucidated. Therefore, we aimed totest whether IKs channels activated by pathological Ca2+ overload may compensate for arrhythmias using
genetically engineered (IKs-Tg) mice expressing cardiac human IKs channels. We employed a sepsis model for
pathological Ca2+ overload condition. We found that the sepsis score IKs-Tg mice was significantly lower than thatin wild-type mice, suggesting a protective role of the IKs channel on cardiac pathological modification by sepsis.

Copyright © The Japanese Pharmacological Society2-B-P-136

一般演題（ポスター）

SARAF and ALG-2 are degraded under the ER stress condition in heart failure〇横江 俊一、朝日 通雄
大阪医科薬科大・医・薬理SARAF is an ER membrane protein, that inactivates store operated calcium entry (SOCE). ALG-2 regulates
SARAF expression by modulating E3 ligases that are known to target SARAF. Although SARAF and ALG-2 areexpressed in the heart, their pathophysiological role remains unknown. Here, we found that SARAF protein
expression level was significantly decreased due to its polyubiquitination in cardiomyocytes excised from dilated
cardiomyopathy (DCM) mice compared to control mice. The ALG-2 expression level was also significantly
decreased in cardiomyocytes excised from DCM mice compared to control mice. We next observed that theexpression level of both SARAF and ALG-2 are significantly decreased in ER stress-exposed HEK293 cells.
Furthermore, we observed that knockdown ofdecreased SARAF expression level in HEK293 cells. These

data indicate that SARAF is degraded via ALG-2-mediated polyubiquitination under the ER stress condition in
failing hearts. Further work is necessary to determine whether SARAF and/or ALG-2 can regulate cardiac
function in DCM especially through in ER stress conditions.

Copyright © The Japanese Pharmacological Society2-B-P-137

一般演題（ポスター）

YAP promotes aerobic glycolysis by upregulating GLUT1 in cardiomyocytes in
response to acute pressure overload〇柏原 俊英1、中原 努1、佐渡島 純一2
北里大・薬・分子薬理、2Rutgers New Jersey Med. Sch., Cardiovasc. Res. Inst., Dept. of Cell Biol. and Mol. Med.

1

Yes-associated protein 1 (YAP), a major transcriptional cofactor in the Hippo signaling pathway, is known toregulate cell growth and homeostasis. We have shown previously that YAP is activated and mediates adaptive
cardiac hypertrophy in response to acute pressure overload (PO). However, how YAP induces adaptive
hypertrophy is unknown. Glycolysis is intimately involved in cell growth, including cardiac hypertrophy. Here
we examined whether YAP regulates glycolysis during acute PO to promote adaptive cardiac hypertrophy.
Evaluation of extracellular acidification rate using the Seahorse XF analyzer in freshly isolated adult ventricular
myocytes (AVMs) from wild-type mice revealed that glycolytic flux was increased by acute PO. The PO-induced
glycolysis was attenuated in isolated AVMs from cardiac-specific heterozygous YAP knockout mice. We foundthat YAP promoted glycolysis by upregulating glucose transporter 1 (GLUT1), which in turn caused
accumulation of glucose metabolites, including L-serine, L-aspartate, and malate, during acute PO. YAP
overexpression increased GLUT1 protein levels, glycolytic flux, and cardiomyocytes size in isolated AVMs.
These results suggest that YAP induces adaptive cardiac hypertrophy through activation of aerobic glycolysis in
response to acute PO.

Copyright © The Japanese Pharmacological Society2-B-P-138

一般演題（ポスター）

TNF-α induces human aortic valve interstitial cell calcification by inhibiting CD34
gene expression〇于 在強1、大徳 和之1、皆川 正仁1、今泉 忠淳2、元村 成3、古川 賢一4、瀬谷 和彦2
1

弘前大・院医・胸部心臓血管外科学、2弘前大・院医・脳血管病態学、3弘前大・院医・病態薬理学、4弘前大・院医・整形外科学

Aortic valve stenosis (AVS) occurs frequently in the elderly, which is exacerbated by valve ectopic calcification.
Recently, we found that mesenchymal undifferentiated cells were rich in human aortic valve interstitial cells(HAVICs), and hematopoietic stem cell marker CD34-negative cells were higher sensitive on various
calcification stimulations such as tumor necrosis factor (TNF)-α. These cells were also vascular endothelial
growth factor receptor 2-positive. We aimed to investigate whether TNF-α affects the gene expression of CD34
and calcification-related proteins in HAVICs, isolated from the calcified aortic valves of AVS patients. After 8hculture of HAVICs with TNF-α (30 ng/mL), the gene expressions of bone morphogenetic protein (BMP) 2
and distal-less homeobox 5 were accelerated. In addition to CD34, gene expressions of extracellular matrix
proteins tenascin X and matrix Gla protein were significantly decreased after 6 days culture of HAVICs with
TNF-α, resulting in accelerated calcification. Transforming growth factor-β1, known another calcificationinducing factor, did not change these gene expressions. These results suggest that TNF-α-BMP2 signaling
mainly contributes to valve ectopic calcification by inhibiting CD34 expression.

Copyright © The Japanese Pharmacological Society2-B-P-139

一般演題（ポスター）

Relevance of characterizing calcitonin gene-related peptide knockout mice in the
Parkinson‘s disease model〇米山 佳和1、孫 煕文2、荒 智大2、橋川 直也1,2、橋川 成美1,2
岡山理科大・理学研究科、2岡山理科大・理1

Depressive disorders occur in 40%‒50% of patients with Parkinsonʻs disease. Calcitonin gene-related peptide(CGRP) is a neuropeptide known as a pain transmitter. Our previous study suggested that
intracerebroventricular administration of CGRP improves depressive-like behaviors. Here, we presented
evidence that CGRP-deficient knockout (CGRP KO) mice exhibit Parkinsonʻs disease symptoms withdepression-like behaviors. We also investigated whether intranasal administration of CGRP affects theamelioration of Parkinsonʻs disease-like symptoms in CGRP KO mice. Saline or CGRP (0.1 mM) was
intranasally administered to 10-week-old CGRP KO mice for 2 weeks. We performed several behavioral
paradigm tests to evaluate the miceʻs motor and cognitive functions: open field, catalepsy, pole, rotarod, hindlimb, and adhesive removal. Motor dysfunction was observed in the CGRP KO mice in all behavioral tests
compared with the wild-type control C57BL6/J mice. Intranasal administration of CGRP ameliorated motor
function in the rotarod test and tended to improve it in the catalepsy test. This administration also increased thelevel of tyrosine hydroxylase in the substantia nigra in the CGRP KO mice. This studyʻs results suggest that
CGRP replacement is a novel therapeutic approach for Parkinsonʻs disease.

Copyright © The Japanese Pharmacological Society2-B-P-140

一般演題（ポスター）

The impacts on fetal brain by maternal hypoxic stress〇徳留 健太郎1、植木 正明1,2、中村 敦輝1、本間 拓二郎1、松永 慎司1、冨田 修平11

大阪公立大・院医・分子病態薬理、2西脇病院・麻酔科

Maternal hypoxic stress such as threatened abortion at early pregnancy is thought to be a risk factor for onset ofchildrenʻs neurodevelopmental disorder. In fact, we previously demonstrated that maternal stress by rearing at
hypoxic condition caused communication deficit and learning disability of rat pups. To clarify the onset
mechanism of neurodevelopmental disorder-like phenotype, we conducted immunohistochemical studies byusing rat fetal brain. Firstly, TUNEL stain revealed that maternal hypoxic stress didnʻt induce cell death. Then,
immunofluorescent double staining for analyzing proliferation and differentiation of neural stem cells (NSCs)
were conducted. NSCs proliferation ability analysis revealed that the rate of Ki67+SOX2+ cells were significantly
increased in fetal rat brain received maternal hypoxic stress. In addition, analysis of asymmetric division whichmeans initial step of neurogenesis, by using SOX2 and TBR2 (intermediate progenitor cell marker) double
staining, showed that maternal hypoxia induced the increase of the number of NSCs. Furthermore, qPCR
results showed that maternal hypoxia reduced astrogliogenesis-related genes and excitatory neurogenesisrelated gene expression. This study indicated that maternal hypoxia influenced the function of NSCs and theirdestination.

Copyright © The Japanese Pharmacological Society2-B-P-141

一般演題（ポスター）

Evaluation of Autosomal Dominant Nocturnal Frontal Lobe Epilepsy Symptoms
in S284L Mutant Transgenic Rats

〇パブラック 晶子1、佐伯 健輔1、笠井 重幸2、村上 惇2、小栗 均2、村澤 寛泰1、小林 洋之1、伊藤 貴博1、加藤 正巳1、平澤 康史1、長瀬 孝彦11

日本バイオリサーチセンター、2オリエンタルバイオサービス

Drug-induced and electroshock-induced animal models have been used in the development of antiepileptic
drugs, but these models are seizure models, not epilepsy models. For the development of antiepileptic drugs, it
is important to develop epilepsy models that mimic the mechanism of seizure generation in humans. Mutationsin the gene encoding the nicotinic acetyl receptor (nAChR) causes autosomal dominant nocturnal frontal lobe
epilepsy (ADNFLE) in humans. In the present study, we evaluated the epileptic phenotype of CHRNA4 S284L
mutant transgenic rats by measuring EEG and EMG, following the report of Zhu et al. We also evaluated theantiepileptic effects of carbamazepine and zonisamide. In addition, cognitive function, anxiety symptoms, andcircadian rhythm were evaluated to assess the animals' psychiatric symptoms. As a result, we confirmed
ADNFLE seizures with symptoms of "paroxysmal arousal," "paroxysmal dystonia," and "wandering" in TG rats.
We also confirmed interictal discharges and confirmed that zonisamide was more effective than carbamazepine
for these abnormal discharges. On the other hand, no cognitive dysfunction, anxiety symptoms, or circadianrhythm abnormalities were observed in TG rats. Although these TG rats did not show abnormal psychiatric
symptoms in this study, they exhibited symptoms similar to those of human epilepsy and were responsive todrugs, suggesting that they may assist in the development of antiepileptic drugs.

Copyright © The Japanese Pharmacological Society2-B-P-142

一般演題（ポスター）

Kaufman oculocerebrofacial syndrome gene, Ube3b, is crucial for the
maintenance of synapse numbers in the young adult brain〇勝部 早紀1、小金澤 紀子1,2、花村 健次1,2、Cuthill Katherine J.3、Ambrozkiewicz Mateusz C.3、川辺 浩志1,2
群馬大・医・薬理、2群馬大・院医・薬理、3ベルリン・シャリテ医科大学 細胞生物学神経科学研究所

1

A large number of patients are suffering from developmental disabilities, including Kaufman oculocerebrofacial
syndrome (KOS). KOS is a severe autosomal recessive disorder characterized by general developmental delay
with intellectual disability, hypocholesterolemia, and seizures.

, an E3 ligase gene, has been reported as

the causative gene for KOS. Our previous report demonstrated a loss of murine
dendritic spines at the age of three weeks, indicating that

increases the number ofis a negative regulator of synaptogenesis forthe excitatory synapses in the early developmental stage. In this study, using brain-specific conditional
knockout (bKO) mice, we investigated the further role ofin the young adult period. Mouse brain

sections were prepared from 9-10 weeks old mice and immunostained with antibodies to Bassoon and Homer1,
respective markers for presynapse and excitatory postsynapse. Images from hippocampal CA1 region wereacquired with 3D Stimulated Emission Depletion (3D-STED) microscopy to estimate the excitatory synapse
numbers. We found that the excitatory synapse density significantly decreased in
the control. Together with our previous reports, our results indicate the novel role ofof synapse numbers in the young adult period.

Copyright © The Japanese Pharmacological SocietybKO as compared toin the maintenance2-B-P-143

一般演題（ポスター）

Short-term intranasal rotenone-administrated mice exhibit multiple prodromal
symptoms in Parkinson‘s disease〇佐藤 元1、豊田 博紀2、野崎 一徳3、佐藤 慶太郎1、片桐 綾乃2、安達 一典1、加藤 隆史2
明海大・歯・薬理、2大阪大・院歯・口腔生理、3大阪大・歯・医療情報1

Parkinsonʻs disease (PD) causes multiple non-motor symptoms as well as distinctive motor deficits. It is wellknown that chemosensory and mild cognitive impairments frequently occur in the early stage of PD, but themechanisms underlying these PD-related prodromal impairments remain unknown. We examined whether
intranasal rotenone-administrated male C57BL/6J mouse was a suitable model for elucidation of PD-related
prodromal/non-motor symptoms. The 1-week rotenone treatment induced chemosensory and conditioned taste
aversion memory impairments without locomotor impairments. Subsequent catecholamine (CA) neuron andfiber loss was observed in the first central nervous system for chemosensory information relays (the olfactory
bulb and nucleus of solitary tract) and the critical brain region of taste associative memory (the dysgranularinsular cortex) in 1-week rotenone-treated mice, respectively. Despite no apparent changes in the number of CA
neuron in the substantia nigra and the ventral tegmental area of 1-week rotenone-treated mice, those of 4-weekrotenone-treated mice were significantly reduced (> 70%) at the fifth week. These results suggest that thismodel employing short-term intranasal rotenone administration will be useful for the elucidating the prodromal
stage of PD pathophysiology.

Copyright © The Japanese Pharmacological Society2-B-P-144

一般演題（ポスター）

Exposure to social defeat stress as juveniles leads to activated brain immune
systems and impaired neuronal morphology〇吉田 樹生1、鈴木 千晴2、濱田 眞里亜2、片田 ひかり2、肥田 裕丈2、毛利 彰宏3、𠮷見 陽2,4、尾崎 紀夫4、野田 幸裕
1,2,4
1

名城大・院薬・病態解析学Ⅰ、2名城大・薬・病態解析学Ⅰ、3藤田医科大・院保健・レギュラトリーサイエンス分野、4名

古屋大・院医・精神医学

Exposure to psychosocial stress (e.g., bullying) in juveniles is a risk factor of stress-related psychiatric disorders
later in life. The exposure to stress activates microglia, which plays an important role in brain immunity, andinduces neuroinflammation. It is unclear to what degree the exposure to psychosocial stress as juveniles isaffected to brain immunity systems and neuronal morphology. The present study was examined expression of
inflammatory cytokines or inflammatory signal-related molecules and neuronal morphology in the prefrontal
cortex (PFC) by using the mice exposed to social defeat stress as juveniles. We found that inflammation or
immune system-related genes of defeated mice were significantly changed, compared to those of non-defeatedmice in transcriptome analysis. Especially, the high expressions of Ca2+ binding protein, S100A8, and S100A9
genes were observed in the PFC of defeated mice. The levels of TNF-α and the numbers of spines in defeatedmice were significantly increased and decreased, respectively, compared to that in non-defeated mice. There
were no significant changes of TNFR1, NF-κB, or I-κB levels in defeated mice. Administration of R-7050, a
TNF-α receptor antagonist, didnʻt develop the impairment of social behaviors induced by social defeat stress
exposure as juveniles. Our findings suggest that exposure to social defeat stress as juveniles induces TNF-α
mediated neuroinflammation via the activated microglia.

Copyright © The Japanese Pharmacological Society2-B-P-145

一般演題（ポスター）

Mass spectrometric analysis of Parkinson‘s disease brain by Py-Tag derivative
reagent.

〇鹿野 仁美1、池田 明夏里2、寺内 勉2、横山 順2、平 修1
1

福島大学・農学群・食農学類、2太陽日酸株式会社・SI事業部

To demonstrate the accurate analysis of catecholamines and amino acid using derivatization reagents, weinvestigated the reaction conditions for 2,4,6-triethyl-3,5-dimethyl pyrylium trifluoromethanesulfonate (PyTag), derivatization of the targets dopamine (DA) and γ-aminobutyric acid (GABA) on tissue sections, andconstructed an optimized reaction compartment. Ten different Py-Tag reaction conditions with the targets wereconsidered. The optimal condition for the Py-Tag reaction with the targets was identified as a 70% methanol
with 5% trimethylamine solution at 60 ℃ under homogenous conditions. To reproduce this reaction on tissue
sections, we constructed a reaction compartment to maintain humidity levels and facilitate the derivatization
reaction. Moreover, visualization of DA and GABA was archived by derivatized-imaging mass spectrometry.
Brain sections of unilateral Parkinsonʻs disease (PD) model rats showed Py-Tag DA in the unilateral striatum
and Py-Tag GABA in the cerebral cortex, striatum, hippocampus and hypothalamus. Using the PD model rat
brain, images with left-right differences were obtained for the localization of DA and GABA. These findings
indicate, it is important to consider the reaction conditions that allow high reaction efficiency between DA orGABA and Py-Tag as well as high quality imaging of sections.

Copyright © The Japanese Pharmacological Society2-B-P-146

一般演題（ポスター）

Sympathetic reactivities by restraint stress change during peri-pubertal stages inrats〇上田 昌史、山口 奈緒子、岡田 尚志郎
愛知医科大・医・薬理学講座

There are differences in stress reactivity between pre-puberty and adult rats. Previous studies showed thatsignificant shifts in the reactivity of hypothalamic-pituitary-adrenal (HPA) axis during the pubertal stages ofdevelopment. Although sympathetic nervous system (SNS) is activated by stressor including restraint stress(RS) in adult rats, it is not well understood whether RS induces the SNS activations in peri-pubertal stages. The
aim of this study is to investigate the age-dependent changes in sympathetic reactivities, as stress response by asingle and repeated RS, in peri-pubertal male rats.
Male Wistar rats at 25-28 postnatal days of age (PND) (pre-puberty) and PND 39-42 (mid-puberty), and PND
84-87 (adult) were used in this study. Rats were exposed to a single RS (1RS) or repeated RS for 4 consecutive
days (4RS). Blood samples were collected immediately after termination of RS. Plasma catecholamines were
measured by HPLC with electrochemical detection to evaluate the SNS activations.
As results, the levels of plasma catecholamines were significantly increased by 1RS in pre-pubertal rats
compared to mid-pubertal and adult rats. In contrast, 4RS did not induce the increasing of the levels of plasma
catecholamines in pre-pubertal rats. These results indicate that the SNS in pre-puberty is more severely
activated by a single RS than that in mid-puberty and adult.

Copyright © The Japanese Pharmacological Society2-B-P-147

一般演題（ポスター）

The analysis of synapse numbers in an autism spectrum disorder (ASD) model
mouse with 3D-STED microscopy〇中村 友哉1、花村 健次1,2、堀 啓3、星野 幹雄3、川辺 浩史1,2
群馬大・医・薬理、2群馬大・院医・薬理、3国立精神・神経医療研究セ・神経研・病態生化学研

1

Autism spectrum disorders (ASD) is a developmental disorder with an estimated prevalence of ~1.6%. Growing
financial and caregiving burdens for patientsʻ families are getting social problems. Diagnostic criteria of ASD
include deficits in social communications and interactions, restricted interests, and repetitive behaviors. Basedon studies of monozygotic and dizygotic twins, it has been proposed that a part of ASD is a genetic disorder.
Indeed, many causative genes of ASD, such as Autism susceptibility candidate 2 (AUTS2) have been identified.
It has been reported that the murine orthologue of AUTS2, Auts2 activates a small GTPase Rac1, and thus
regulates neuronal migration by forming lamellipodia at the growth cone. Given that lamellipodia play crucial
roles in synapse formation as well, we estimated synapse numbers in Auts2 mutant mouse in this study.
Excitatory synapses were immunolabeled by antibodies to Bassoon and Homer1, markers for pre-synapse andexcitatory post-synapse respectively. Synapses at hippocampal CA1 region were imaged with super-resolved
three-dimensional stimulated emission depletion (3D-STED) microscopy. As a result, we discovered the changeof the number of excitatory synapses in Auts2 mutant mouse. Our finding sheds light on unidentified
pathological changes in ASD at the synapse level.

Copyright © The Japanese Pharmacological Society2-B-P-148

一般演題（ポスター）

Investigating the effects of ultrasound exposure on emotion using animalmodels〇山内 つぐみ1,2、吉岡 寿倫1、山田 大輔1、濱野 匠1、公木 彩夏1、入山 聖史3、吉澤 一巳4、市川 寛子2、西野 彰一5、
宮崎 智6、斎藤 顕宜11

東京理科大・薬・薬学科、2東京理科大・教養教育研究院・心理学研究室、3東京理科大・理工・量子情報力学研究室、4

東京理科大・薬・疾患薬理学研究室、5株式会社フジミック、6東京理科大・薬・生命情報科学研究室

Recently, ultrasound exposure has been shown to be a noninvasive method for modulating brain activity and has
to be applied to treat mental disorders, such as depression. However, its underlying mechanism remains unclear.
Therefore, there is a need for animal models that can investigate the mechanism of ultrasound exposure. In thisstudy, we utilized olfactory bulbectomized (OB) rats as an animal model of depression and investigated theiremotional state following ultrasound exposure. As a result, following exposure to high-frequency ultrasonicvocalizations (USV)s of rats, the hyperemotionality of OB rats was significantly suppressed. Also, USV exposure
significantly reduced the latency to the first entry into the open arm in the elevated plus maze tests and theplasma corticosterone levels of OB rats. Furthermore, artificial ultrasounds (50 and 100 kHz) also significantly
decreased the hyperemotionality score of OB rats. These results suggested that ultrasound exposure, regardlessof vocal or artificial sound, ameliorated depressive-like behavior and stress level in OB rats. We proposed that
OB rats might be an appropriate animal model to identify mechanisms underlying the effects of ultrasoundexposure.

Copyright © The Japanese Pharmacological Society2-B-P-149

一般演題（ポスター）

The effects of DOP agonists on the extinction and reconsolidation of fear
memory in mice.

〇河南 絢子1、山田 大輔1、栁澤 祥子1、白方 基揮1、畠山 梓摘1、飯尾 啓太2、長瀬 博2、斎藤 顕宜11

東京理科大・薬・薬理学研究室、2筑波大・国際統合睡眠医科学研究機構

Facilitation of fear extinction and inhibition of reconsolidation of fear memory are useful for the treatment offear-related disorders, such as post-traumatic stress disorder (PTSD). Here, we investigated the effects of two
DOP agonists, KNT-127 and SNC80, with different chemical structures on the fear extinction andreconsolidation of fear memory in mice. On day 1, male C57BL/6J mice were contextually conditioned with 8
footshocks. On day 2, the mice were re-exposed to the conditioning chamber as a memory retrieval session (reexposure 1). DOP agonists were administered subcutaneously 30 min before (for extinction) or immediately
after (for reconsolidation) the re-exposure 1. On day 3, mice were re-exposed to the chamber as a memory
testing session (re-exposure 2). The duration of re-exposure was 6 min for extinction and was 2 min forreconsolidation. As a result, KNT-127 (3‒10 mg/kg), but not SNC80 (1‒10 mg/kg), administered to the micefollowing both 6-min and 2-min re-exposure 1 significantly decreased the freezing rates of mice during reexposure 2. These effects of KNT-127 were abolished by pretreatment with a selective DOP antagonist. In
conclusion, we propose that KNT-127 facilitates fear extinction and inhibits reconsolidation of fear memory via
DOP.

Copyright © The Japanese Pharmacological Society2-B-P-150

一般演題（ポスター）

Prolonged, but not single, administration of nandrolone inhibits morphineinduced increases in accumbal dopamine efflux in rats

〇川島 央暉1、青野 悠里1、榛葉 繁紀2、三枝 禎1
1

日本大・松戸歯学部・薬理学、2日本大・薬・健康衛生

Nandrolone (NDL) is an anabolic androgenic steroid known to be misused by elite athletes as well as someadolescents due to its muscle-building properties. We have shown that a 4-week period of repeatedadministration of NDL to 6-, but not 10-week-old rats enhances the developmental increase in grip strength
without influencing developmental increases in body weight. Furthermore, systemic administration of morphine
induced a smaller increase in dopamine (DA) efflux in the nucleus accumbens (NAc) of rats that had received
NDL treatment for 4 weeks relative to those without NDL treatment. Recently, a single administration of NDL
to mice was shown to enhance DAergic neural firing in the ventral tegmental area (VTA: Bontempi & Bonci,
2020). The DAergic neurons in the VTA are known to send projections to the NAc. Accordingly, we further
analyzed the effects of a single administration of NDL on baseline as well as morphine-induced increases inaccumbal DA efflux using

microdialysis. Male Sprague-Dawley rats approximately 10 weeks old wereused. A single administration of NDL (5.0 mg/kg s.c., 24 hr before morphine treatment) did not alter either
basal levels of accumbal DA or the increase in accumbal DA efflux induced by systemic administration ofmorphine (1.0 mg/kg, s.c.). These results, together with our previous findings, show that prolonged but not asingle administration of NDL in rats can inhibit morphine-induced increases in accumbal DA efflux.

Copyright © The Japanese Pharmacological Society2-B-P-151

一般演題（ポスター）

Labeling method for memory-related synapses〇大西 泰地1、坂本 寛和1、大久保 洋平2、並木 繁行1、廣瀬 謙造1
東京大・院医・細胞分子薬理学、2順天堂大・医・薬理1In the auditory cued fear conditioning, memories are stored in synapses between engram neurons, which
increased activity during the memory formation. Although molecular imaging of memory-related synapses
contributes to deciphering the molecular basis of memory, there is no established method for visualizing them.
In this study, we developed a method to selectively visualize synapses between engram neurons, using the c-fos
promoter-driven Tet-On system. Tag-fused synaptophysin (tagSyp) and FingR.PSD95 were expressed inpresynaptic auditory cortex neurons and postsynaptic lateral amygdala neurons, respectively, in an activitydependent manner. We found that the number of synapses positive for both tagSyp and FingR.PSD95 in thelateral amygdala was 15-fold higher in mice with cued fear conditioning than in control mice. Thus, weconcluded that synapses both positive for tagSyp and FingR.PSD95 correspond to memory-related synapses.
Furthermore, combining this method with immunohistochemistry, we found greater amounts of several kinds ofsynaptic molecules accumulate in memory-related synapses compared to othersynapses. It is expected that our

method enables comprehensive analysis of molecular changes in synapses induced by fear memory formation.

Copyright © The Japanese Pharmacological Society2-B-P-152

一般演題（ポスター）

Changes in spontaneous firing frequency of striatal cholinergic interneurons inaged mice.

〇鈴木 江津子、籾山 俊彦
東京慈恵会医科大・医It has been reported that frequency of action potential firing of striatal cholinergic interneurons increases withpostnatal development. On the other hand, changes in spontaneous firing frequency and firing properties during
aging have not been investigated. In this study, cell-attached and whole-cell patch-clamp studies were carriedout to investigate changes in firing properties of striatal cholinergic interneurons during aging. Brain slices wereprepared from 2‒3-month-old, 11‒12-month-old and 24-month-old mice of either sex. Frequencies of
spontaneous firing at 2‒3-month-old, 11‒12-month-old and 24-month-old were 4.55 ± 1.01 Hz (n = 18), 8.73
± 2.28 Hz (n = 8) and 2.88 ± 0.99 Hz (n = 11), respectively. Firing frequency at 14 month of age wassignificantly decreased than that of 11‒12-month-old (p = 0.028). Since spontaneous firing appeared to beirregular in 24-month-old mice, the coefficient of variation (CV) of the inter-event interval of spontaneous
firing was analyzed. CV at 24 months of age (0.68 ± 0.1, n = 10) was significantly larger than that of 2-3-monthof age (0.34 ± 0.05, n = 18, P = 0.015), indicating that the regular pattern of spontaneous firing is disrupted in
24-month-old mice. These findings suggest prominent changes in firing properties of striatal cholinergic
interneurons during aging.

Copyright © The Japanese Pharmacological Society2-B-P-153

一般演題（ポスター）

Increased neuronal activity of serotonin neurons in the median raphe nucleus
attenuates reward-related consummatory responses and facial expression〇大村 優1、ブシェキワ ユセフ1,2、河合 洋幸3,4、永安 一樹3、金子 周司3
北海道大・院医、2City University of New York・Queens College・Dept. Psychology、3京都大・院医薬・生体機能解析、41

大阪公立大・院医・脳神経機能形態学

We have previously demonstrated that serotonin neurons in the median raphe nucleus (MRN) facilitate anxietyand fear-related behavior evaluated by elevated plus maze and fear conditioning tests, respectively. However, we
still cannot conclude that the neuronal activity of serotonin neurons in the MRN directly causes negative
emotion because the time gap between the experimental manipulations and the evaluation of resulting
behavioral changes is relatively long in these behavioral tests. In other words, learning or planning processes
could affect performance in these tests. To address this issue, we optogenetically stimulated serotonin neurons
in the MRN and evaluated quick responses to sucrose reward in mice. We delivered a sucrose reward from ablunt-tipped needle placed at a licking distance and measured mice licking behavior. The optogenetic activationof serotonin neurons in the MRN reduced the number of licks. Furthermore, to assess hedonic responses more
directly, we intraorally infused a drop of sucrose solution and analyzed taste reactivity and facial expression. We
found that activating serotonin neurons in the MRN significantly reduced the number of tongue reactions andsweet sucrose-driven facial expressions. These results suggest that increased activity of serotonin neurons in the
MRN mediates aversion.

Copyright © The Japanese Pharmacological Society2-B-P-154

一般演題（ポスター）

Histamine neurons promote the recall of associative memories〇髙村 侑希1、西村 京華1、人羅（今村） 菜津子1,2、南 雅文1、野村 洋1,3
北海道大・薬・薬理学研究室、2熊本大・院生命科学、3名古屋市立大・院医・認知機能病態学寄附講座

1

In the brain, histamine is produced mainly in the tuberomammillary nucleus (TMN) and involved inlearning/memory, sleep/wakefulness, feeding, and neuroendocrine regulation. Previously, we demonstrated that
activation of the histaminergic neurotransmission by histamine H3 receptor inverse agonists promotes the recallof forgotten memories. However, it remains unclear whether histamine neurons are activated during memory
recall and whether activation of histamine neurons is required for memory recall. In this study, first, we usedfiber photometry to measure the activity of histamine neurons. We introduced GCaMP6s into histamine
neurons of HDC-Cre mice and acquired the fluorescence via optic fibers. The mice were subjected to auditoryreward conditioning. The activity of histamine neurons increased when the mice recalled the reward memory by
the conditioning tone. Next, we used optogenetics to test whether the activation of histamine neurons is
involved in memory recall. We introduced NpHR into histamine neurons of HDC-Cre mice and inhibited theiractivity. Inhibition of histamine neurons reduced tone-induced licking behavior that is based on recall of reward
memory associated with auditory stimulus. These results suggest that histamine neurons contribute topromoting the recall of associative memories.

Copyright © The Japanese Pharmacological Society2-B-P-155

一般演題（ポスター）

Effects of systemic inflammation and histamine on the network oscillation in theanterior cingulate cortex〇村越 隆之1、平尾 鮎美1、村上 元2、伊藤 吏那1、魚住 尚紀1
埼玉医科大・医・生化、2埼玉医科大・医・教養教育

1

Since the arousal and cognition are considered to be maintained by the network oscillation generated by neuralcircuits in the anterior cingulate cortex (ACC), and are deeply influenced by histaminergic system and disturbed
under the systemic inflammation, it is expected that elevated inflammatory cytokines would alter ACC network
oscillations along with the behavioral changes. We studied the effects of lipopolysaccharide (LPS) on ACCrelated task and kainic acid (KA) -induced network oscillations in an ACC slice, to search the relationshipamong LPS-induced elevation of cytokine level, the profiles of behavior and network oscillations.
<Materials & Methods> Male C57BL/6J mice were intraperitoneally administered with LPS (30 µg/kg). Threehours later, novel object recognition test (NOR) was performed, followed by preparations of blood and corticaltissue samples, and brain slices containing ACC for electrophysiology.
<Results> LPS treatment significantly increased serum IL-6 level and tended to increase brain tissue IL-6
level. Network oscillation was evoked by KA (0.3-3 µM) perfusion alone or with histamine (10 µM) as fieldpotential activities within the ACC. Power of KA-induced oscillation increased in the LPS groupcompared with the control. Histamine enhanced oscillation power in the control group, but this was notobserved in the LPS group. In the NOR, exploration time for a novel object decreased in the LPS group but
without a significant difference in the recognition index between the two groups.

Copyright © The Japanese Pharmacological Society2-B-P-156

一般演題（ポスター）

Region-specific plastic changes in the prefrontal cortex drive compulsive task
performance〇浅岡 希美、林 康紀
京都大・院医・システム神経薬理

Compulsive actions, defined by maladaptive perseveration and excessive repeat, are shown in various mental
diseases. Recent evidence suggests that habit-based decision-making causes such perseveration; however, neuralmechanisms for developing habitual, and also excessive, action remain unclear. Here, we developed an original
operant overtraining protocol that induces habitual and excessive action in mice. Such compulsive-like action is
only observed in about half of overtrained mice, whereas other mice show habitual but not excessive action.
During the course of training, training-induced plastic changes in excitatory synaptic inputs, as assessed by
AMPA/NMDA ratio, was observed in several regions of the prefrontal cortex. As mice learned the task, anincrease in AMPA/NMDA ratio was observed in layer 5 pyramidal neurons of the anterior cingulate cortex(ACC) and lateral orbitofrontal cortex (OFC). When mice were overtrained on a slightly modified task for
additional 4 days, the increase in AMPA/NMDA ratio was no longer observed in the ACC of overtrained mice
regardless of the excessiveness, whereas the increase was only detected in the lateral OFC of mice showing both
habitual and excessive action. Consistently, chemogenetic activation of ACC neurons of overtrained mice
restored goal-directed action and inhibition of lateral OFC neurons of compulsive-like mice suppressed
excessive response. These results indicate that region-specific changes in excitatory inputs in the prefrontal
cortex are critical for the development of compulsivity.

Copyright © The Japanese Pharmacological Society2-B-P-157

一般演題（ポスター）

Evaluation of addiction-inducing drugs based on the electrical activity of human
iPS cell-derived dopamine neurons〇石橋 勇人、永福 菜美、鈴木 郁郎
東北工業大

Addiction is known to be caused by various compounds such as frequently used medicines, luxury goods, andillegal drugs, and there is a need to establish an evaluation system for side effects to prevent the formation of
addiction. However, a method for evaluating addiction has not yet been established. In this study, weconstructed an evaluation system to detect addiction-positive compounds by measuring the electrical activity of
iPS cell-derived dopamine neurons using microelectrode arrays (MEAs). The dopamine neurons (iCell) andastrocytes (iCell) were co-cultured on MEA. After 35 days of culture, we conducted a chronic administration
test of 10 addiction-related compounds and the results before and after the chronic administration werecompared using principal component analysis (PCA). In the cumulative administration experiment using
dopamine neurons, it was possible to detect different responses for addiction-positive and addiction-negative
compounds acting on the same receptors. In the chronic administration test, four out of five compounds weredetected for the addiction compound, and the non-addiction compound was not misclassified as an addictive
compound, suggest that this evaluation method is capable of detecting the addiction compound. These results
suggest that the evaluation system used in this study is useful as a screening system for addiction-positive
compounds.

Copyright © The Japanese Pharmacological Society2-B-P-158

一般演題（ポスター）

Our pre-clinical support for neurotransmitter evaluation in the brain of rodents
using a microdialysis method〇荒木 康平、齋藤 慶太、鈴木 孝太郎、野々村 徹、山崎 則之
株式会社新薬リサーチセンター・非臨床研究部【Objective】
In vivo microdialysis is a technology to measure the concentration of a target substance in extracellular fluid at a
localized site by collecting the substance through a dialysis membrane. In the brain, the dialysate enables us toassess real-time changes in neurotransmitters. Therefore, the technique is a useful method for thepharmacological action of central nervous system compounds. In order to fully support researchers, we would
like to present an example of the antidepressant evaluation systems using the dialysate from various brain
regions in rodents.
【Methods & Results】
A microdialysis probe was inserted into each of the hippocampus or the medial prefrontal cortex (mPFC) in
C57BL/6J mice. After 2hour of pre-perfusion, the dialysate was collected at 20-minute intervals at a flow rate of
1µL/min. Then, mice were treated with intraperitoneal injection of R-fluoxetine, a selective serotonin reuptake
inhibitor for the treatment of depression, and the dialysate was recollected. The next day, dopamine andserotonin levels were measured by HPLC-ECD. As a result of the study, serotonin release was significantly
increased in the hippocampus compared to pre-administration. On the other hand, dopamine release wassignificantly increased in mPFC compared to pre-administration.
【Conclusions】
In this study, we confirmed the effects of a single antidepressant administration on brain monoamines in wildtype mice. We will continue to introduce a wide range of technologies and evaluation systems to provide valueadded

clinical testing services.

Copyright © The Japanese Pharmacological Society2-B-P-159

一般演題（ポスター）

Roles of brain carbon monoxide in micturition of rats

〇山本 雅樹1、清水 孝洋2、Zou Suo2、清水 翔吾2、東 洋一郎2、藤枝 幹也1、齊藤 源顕2
高知大・医・小児思春期医学、2高知大・医・薬理学講座

1

Endogenous carbon monoxide (CO) produced by heme oxygenase (HO) is reported as a relaxation factor in theurethral smooth muscle, while roles of CO in the brain in regulation of the micturition reflex remain unclear. In
this study, to elucidate roles of brain endogenous CO in regulation of the micturition reflex, we investigatedeffects of centrally administered CORM3 (CO donor) and ZnPP (non-selective HO inhibitor) on themicturition reflex in urethane-anesthetized (0.8 g/kg, ip) male Wistar rats. A catheter was inserted into thebladder to perform cystometrograms (CMG), which was started 2 h after the surgery, and 1 h after the start,
CORM3 or ZnPP was intracerebroventricularly (icv) administered. CORM3 (1 or 10 nmol/rat, icv) dosedependently prolonged intercontraction intervals (ICI), while ZnPP (10 or 30 nmol/rat, icv) dose-dependently
shortened ICI. The ZnPP (30 nmol/rat, icv)-induced ICI shortening was reversed in the presence of CORM3(10 nmol/rat, icv). In addition, ZnPP (30 nmol/rat, icv) significantly reduced single-voided volume and bladdercapacity without affecting post-voiding residual volume or voiding efficiency. These results suggest that brain
endogenous CO can inhibit the micturition reflex in rats.

Copyright © The Japanese Pharmacological Society2-B-P-160

一般演題（ポスター）

TRPC3/6 channels inhibitor L862 exhibits protective effect against PAN-induced
cellular damage in mouse podocyte〇松田 由宗1,2、坂口 怜子1、岡田 亮1,3、木原 隆典2、永田 龍4、森 誠之11

産業医科大・医・医学科生命科学分野生体物質化学、2北九州市立大学・国際環境工学研究科・環境システム専攻、3

産業医科大・医療保健・人間情報科学、4大阪大・院薬Transient Receptor Potential Canonical (TRPC) 6 is highly expressed in glomerular epithelial cells (podocytes)
and is known to contribute to the maintenance of glomerular protein filtration. It has been reported thatpatients with nephrotic syndrome, which is a disease characterized by proteinuria due to renal glomerular
damage, show elevated expression of TRPC6 that causes the disorganization of actin filaments in podocytes. It
has also been reported that gain-of-function mutations of TRPC6 are associated with nephrotic syndrome.
Therefore, specific inhibition of TRPC6 would be an attractive strategy for suppressing proteinuria.
In this study, we have identified a novel TRPC3/6 channels inhibitor, L862. To evaluate the effect of L862 on
podocytes, MPC-5 (Mouse Podocyte Clone 5) were treated with L862. Comparison of the viability of L862treated group to non-treated group showed no significant difference, indicating that L862 does not have toxicity
at the cellular level. Next, we prepared MPC-5 cells treated with Puromycin Aminonucleoside (PAN), which is
known to cause podocyte damage. Treatment of L862 to these damaged MPC-5 exhibited protective effect oncell morphology. These results suggest the potential of L862 as a drug for suppression of proteinuria.

Copyright © The Japanese Pharmacological Society2-B-P-161

一般演題（ポスター）

Utilizing split-luciferase-based HTS platform and natural product extracts libraryyielded cyclosporin A as candidate drug for Alport syndrome〇スイコ メリーアン、桑水流 淳、大町 紘平、小嶋 遥、加世田 翔大、首藤 剛、甲斐 広文
熊本大Alport syndrome (AS) is a hereditary kidney disease caused by mutation in type IV collagen alpha (Col4A) 3, 4,
5 chains, which disrupts trimerization, leading to kidney dysfunction. Mutations in Col4A5 comprise more than
80% of AS-associated mutations. These mutations can hinder trimer formation and/or trimer secretion.
Correcting the trimerization and secretion of Col4A3/4/5 is a feasible therapeutic approach, but is hampered by
the absence of high-throughput screening (HTS) platforms for assessment. We previously created an HTS
system based on split nanoluciferase in which Large BiT (LgBiT) or Small BiT (SmBiT) subunits were fused to
Col4A monomers. Proper trimerization results in complementation of LgBiT and SmBiT to produce
quantifiable luminescence. Here, we used this system to screen natural product extracts library for AS drugcandidate. For screening, we established HEK293T cells stably expressing wild-type Col4a3-SmBiT, Col4a4 andCol4a5 G1244D mutant-LgBiT. The library we screened is composed of extracts from fungi, bacteria, marine
sponge and plants (>15,000 extracts screened). The hit extracts were subjected to mass spec that revealed
cyclosporin A (CsA) as the main component of the extracts. CsA increased the trimer secretion but not trimer
formation of Col4a3/4/5. The cyclophilin binding domain of CsA is important for its secretion-inducing
activity, with cyclophilin D/PPIF being involved in the effect of CsA. Overall, we found a compound that canenhance the secretion of mutant Col4A trimer as candidate therapeutic for AS.

Copyright © The Japanese Pharmacological Society2-B-P-162

一般演題（ポスター）

Synthesis and evaluation of activities of new pantetheine derivatives〇細畑 圭子1、米山 弘樹2、金 徳男3、宇佐美 吉英2、髙井 真司31

大阪医科薬科大・薬・臨床薬学教育研究センター、2大阪医科薬科大・薬・有機薬化学研究室、3大阪医科薬科大学大学院・医学研究科・創薬医学研究室

Background: Acute kidney injury (AKI) is associated with incomplete recovery after the onset of the disease.
However, there is no fundamental treatment for this disease. Previously, we identified vanin-1, which appears on
the cell surface of proximal tubules early after oxidative stress and induces various inflammatory responses. In
this study, we developed and evaluated vanin-1 inhibitors. We synthesized novel vanin-1 inhibitors (OMP
compounds) by chemical modification of pantetheine analogues RR6 (IC50 = 0.54 μM), which was reported by
Schalkwijk J et al. To evaluate the candidate compounds, vanin-1 activity in serum and renal tissues collected at
1 and 4 hours after subcutaneous administration of 10 mg/kg of RR6 or OMP7 to hamsters was compared withthat in the normal group. Results/Conclusions: Serum vanin-1 activity was significantly decreased at 1 hour
after subcutaneous administration of RR6, but there was no significant difference at 4 hours. On the other hand,
vanin-1 activity was significantly suppressed at 1 and 4 hours after subcutaneous administration of the OMP7.
Vanin-1 activity in renal tissue was also not inhibited at 1 hour after subcutaneous administration of RR6. In
contrast, the OMP7 significantly inhibited vanin-1 activity at 1 hour after subcutaneous administration. These
results suggest that compared to RR6, the OMP7 has a sustained inhibitory effect on vanin-1 activity in serum,
and a certain inhibitory effect can be expected in renal tissues as well.

Copyright © The Japanese Pharmacological Society2-B-P-163

一般演題（ポスター）

Eucommia leaf extract improves renal impairment and vascular endothelial
dysfunction in a type 2 diabetes mellitus rat model.

〇中川 恵輔1、中川 愛海1、田中 雅美1、石川 梨乃1、犬塚 理奈1、平田 哲也2、松村 靖夫1、大喜多 守1
大阪医科薬科大・薬・病態分子薬理、2小林製薬・中央研

1

Renal impairment and macrovascular endothelial dysfunction complicated with diabetes mellitus (DM) play amajor role in the life expectancy of DM patients.

leaf extract (ELE) has anti-diabetic effects, but its

effect on those complications with DM are unclear. Therefore, this study used the Zucker diabetic fatty (ZDF,
type 2 DM) rat to examine the effects of ELE-containing diet (3% or 5%) ingestion before or after the onset of
DM. Chronic ingestion of ELE prior to the onset of DM significantly suppressed the elevated blood glucose(BG) levels observed in ZDF rats and also remarkably improved glucose tolerance based on the results of theoral glucose tolerance test. Urinary protein excretion and albuminuria as important indicators of diabetic
nephropathy (DN) were markedly suppressed by taking ELE 5% containing diet. A significant decrease in theacetylcholine-induced vasorelaxation response associated with DM was considerably improved by ELE
ingestion. ELE ingestion after the onset of DM showed a reduction in BG levels, but no obvious effect on DN
and endothelial dysfunction. These results indicate that chronic ingestion of ELE initiated before the onset of
DM not only suppresses BG levels associated with DM progression but also has beneficial effects on DN andvascular endothelial dysfunction.

Copyright © The Japanese Pharmacological Society2-B-P-164

一般演題（ポスター）

Evaluation of urination in a rat cystitis model of induced by hydrogen peroxide〇清水 広夢、森田 枝美、吉原 佐江子、真壁 大地、水町 凉治、田代 貴士、片山 誠一、廣中 直行、西 勝英㈱ＬＳＩＭ安全科学研究所・熊本研究所・薬理研究部

Few interstitial cystitis models can be used for evaluation over a long term although various animal models andevaluation methods exist for the development of novel clinical drugs for treatment of dysuria medicine.
Currently, we produced the acetic acid, hydrochloric acid or cyclophosphamide‒induced cystitis models, but it isdifficult to evaluate the efficacy in the interstitial cystitis model since the symptoms of pollakiuria for all themodels recover in a few days. Therefore, we tried to establish a cystitis model in which frequent urination
symptoms continue for a long period of time using hydrogen peroxide (HP).
Female SD rats were used in the present study. First, we tested both the induction time of HP and themaintenance period of the model , comparing with those in the test groups of a sham group and the groups inwhich HP was stored in the bladder for 5, 15, or 30 minutes. The model rats were prepared by injection of 3%
HP into the bladder under isoflurane inhalation anesthesia, and uroflowmetry was measured at 1, 2, 3, and 4
weeks after establishment of the model.
Results showed that since HP stored in the bladder for 5 minutes did not shorten the micturition interval andsome rats died when HP was stored in the bladder for 30 minutes. Therefore, the model was prepared bystorage of HP in the bladder for 15 minutes when a micturition interval was shortened. The micturition interval
was found to be shorten up to 4 weeks in the uroflowmetry measurement. This model is considered useful for along period of time for cystitis evaluation.

Copyright © The Japanese Pharmacological Society2-B-P-165

一般演題（ポスター）

An estimation of urine flow rate using urinary creatinine excretion rate in rats

小口 茜、〇園田 紘子、川口 珠実、池田 正浩
宮崎大・農Background:In humans, urine collection is accessible because of their altruistic behaviors. On the other hand, since animals
urinate spontaneously, urine collection is difficult. In this study, because it has been known that the urinary
creatinine excretion rate is almost constant in normal animals of similar age of the same species, we try toestimate the urine flow rate using the excretion rate in normal age-matched rats.
Methods and Results:
First, using urine collected from normal male SD rats, the urinary creatinine excretion rate was calculated by
dividing the total urinary creatinine amount by the collection time, and the resulting average value was 8.95
micro g/min. Next, we calculated an estimated urinary flow rate (eUF) using the value mentioned above andurinary creatinine concentration in each individual, and compared the resulting eUF with an actual urinary flowrate (aUF) obtained by considering collected urine volume and time. When the body fluid balance of rat wasaltered by changing an amount of drinking water or a diuretic-treatment, a high positive correlation wasobserved between the aUF and eUF for either a change in water intake (r = 0.90) or a diuretic treatment group(r = 0.77). A similar result was obtained, when we calculated free water clearance using aUF and eUF,
respectively (r = 0.89).
Conclusion:
These results suggest that the urine flow rate can be estimated from the average urinary creatinine excretion
rate in normal rats. In the future, animal species differences and individuals with altered urinary creatinine
excretion rate should be examined.

Copyright © The Japanese Pharmacological Society2-B-P-166

一般演題（ポスター）

Identification of efficient signal peptide in extracellular secretions for mRNA
vaccine development〇皆川 直樹、平田 悠朗、金子 雅幸、岡元 拓海
長崎大・院医歯薬・創薬薬理学

In mRNA vaccines, mRNA encoding antigens enclosed in lipid nanoparticles (LNPs) are injected into themuscle to induce intracellular antigens, which are then secreted extracellularly and recognized by antigenpresenting cells, thereby conferring immunity. Therefore, if the extracellular secretion of antigen is increased,
immunity can be efficiently acquired with a smaller amount of mRNA; thus contributing to the improvement ofvaccine supply.
The secretory efficiency ranking of artificial and natural signal peptides has already been reported. In this study,
we transfected HEK293 cells with a vector in which the N-terminal IL-6 signal peptide of NanoLuc® luciferase(Nluc) was replaced by the signal peptides with high secretory signal strength. The amount of Nluc secreted
extracellularly at each signal peptide was determined by luciferase assays and Western blotting. As per the
results, Nluc with the signal peptide of Cystatin S, a natural signal peptide, had the highest extracellularsecretion. The Western blotting result was consistent with that of the luciferase assay. Our future studies will
test those peptides on C2C12 skeletal muscle cells and HepG2 liver cells. We also plan
peptides using mRNA-LNP.

Copyright © The Japanese Pharmacological Society

validation of those2-B-P-167

一般演題（ポスター）

Elimination of Volatile Organic Compounds from indoor air by chemical filter
significantly delays the development of atopic dermatitis in the mice model.

〇大平 智春1、冨田 賢吾2、金木 真央1、早川 千春1、栗原 隆2、高木 哲3、福山 朋季1
1

麻布大・獣医・薬理、2清水建設・技研・医療環境、3麻布大・獣医・小動物外科学

Allergic diseases including atopic dermatitis (AD) and allergic asthma are multifactor diseases, and thepathogenic mechanism is not fully understood yet. The environmental factor is one of the major contributors forallergy development, and several reports demonstrated that Volatile Organic Compounds (VOCs) are an
exacerbating factors for allergy. In this study, we aim to examine the efficacy of elimination of VOCs by chemical
filter in development of atopic dermatitis using a mice model.
A mouse model of hapten-induced AD were exposed to normal air and chemical filtered air (CF) with VOCs
removed. There was a significant decrease in trans epidermal water loss (TEWL) 8 days after induction
compared to the control group, but thereafter there was no change in the control group. Significant reductions
in T and B cell counts by flow cytometry and pathology were observed in the CF group. The pathology of thecontrol and CF groups became comparable in TEWL, skin thickness, and immune-related factors as the days
passed. In study, the fact that significant effects were observed after the onset of the symptoms suggests thatchemicals may be involved where they cause symptoms, furthermore, the impact of VOCs removal in the earlystages of disease onset is currently under investigation.

Copyright © The Japanese Pharmacological Society2-B-P-168

一般演題（ポスター）

Volatile organic compounds below guideline values in hospital facilities affectthe development of pathology in mouse models of lung disease〇冨田 賢吾1、大平 智春2、金木 真央2、早川 千春2、矢野 慧一1、栗原 隆2、高木 哲3、福山 朋季2
清水建設・技研・医療環境、2麻布大・獣医・薬理、3麻布大・獣医・小動物外科

1

Indoor air quality (IAQ) has been suggested to affect respiratory diseases. Large amounts of volatile organic
compounds (VOCs), emitted from indoor building materials, are known to cause healthy people to induce sickbuilding syndrome., thereby being regulated as an IAQ guideline. However, the effects of VOCs below the IAQ
guideline values on disease pathogenesis have not been adequately studied. If indoor VOCs affect diseasedevelopment, they may also influence the treatment of respiratory diseases. In this study, we aimed to clarify theeffects of indoor VOCs on the diseases of allergic asthma and acute lung injury (ALI) in model mice. We
compared the pathological conditions of mice exposed to indoor air containing VOCs below the guideline values
and those exposed to clean air whose VOCs were removed by a chemical filter. In addition, we compared theanimal experimental environment with the clinical environment by collecting the IAQ data from the mouse
breeding environment and the medical facility.
Our results indicated that clean air exposure significantly improved SpO2, histological abnormalities, and gene
expression of pro-inflammatory cytokines in asthma and ALI models. Our findings suggest that indoor VOCs
even below the guideline values can influence the lung and immune function in asthma and ALI.

Copyright © The Japanese Pharmacological Society2-B-P-169

一般演題（ポスター）

Generation of a DNA-aptamer targeting human galectin-7 as a lesion indicator forcholesteatoma〇劉 爽1、竹政 絵理香1、鈴木 康之2、羽藤 直人3、茂木 正樹1
愛媛大・院医、2済生会松山病院・麻酔、3愛媛大・院医・耳鼻咽喉・頭頚部外1

Aiming at complete excision of cholesteatoma during trympanomastoidectomy and therefore reducing the riskof recurrence, the current study was undertaken to develop a seed DNA-aptamer-based fluorophore-probe,
which targets human galectin-7, as an intraoperative lesion-identifying indicator for the surgical treatment ofcholesteatoma. A galectin-7-targeted DNA-aptamer library was generated for labeling the cholesteatoma matrix
using cell-based systematic evolution of ligands by an exponential enrichment technique. The binding
characteristics of the identified aptamers were analyzed, and structure optimization of the identified aptamers
was carried out both in silico and in vitro. Using galectin-7-aptamer guided molecular imaging, the excision
margins of cholesteatoma matrix and surrounding normal tissue were successfully observed in a xenograftedcholesteatoma model. It is highly expected that specific galectin-7-aptamers could progress to future clinical
trials for both imaging and therapeutic applications and therefore benefit cholesteatoma patients.

Copyright © The Japanese Pharmacological Society2-B-P-170

一般演題（ポスター）

Topical treatment with high concentration of ozone water ameliorates
inflammatory responses and breakdown of the cutaneous barrier in a mousemodel of atopic dermatitis through antibacterial effect toward staphylococci〇金木 真央1、大平 智春1、高橋 美優1、内山 淳平2、阿野 哲也3、福山 朋季1
1

麻布大・獣医・薬理、2岡山大・院医歯薬・病原細菌、3伯東

We focus on the ozone water to develop a novel treatment approach for staphylococci related inflammatory skindiseases. In the 95th Annual Meeting of the Japanese Pharmacological Society, we reported the anti-allergic andanti-microbial effect of 3 mg/L ozone water in a mouse model of atopic dermatitis (AD) and staphylococci,
however, its effect was far from clinical application. The aim of this study is to evaluate the anti-microbial andanti-inflammatory property of high concentration (11 mg/L) of ozone water to advance our project. Ozone
water showed a significant bactericidal effect against
Furthermore,,andinduced IL-1β and IL-6 secretion in human epidermal keratinocytes was

significantly inhibited by pre-treatment of ozone water.

experiment with a mouse model of AD,

significant improvement of AD symptoms and trans epidermal water loss were also found in an ozone water
treatment group. Surprisingly, local immune reactions such as type II conventional dendritic cells, effector
helper T cells, and IgE-produced B cells are also regulated by ozone water application. Our findings strongly
suggest that topical treatment with high concentration of ozone water significantly ameliorated AD symptoms
through antibacterial effect toward staphylococci.

Copyright © The Japanese Pharmacological Society.

2-B-P-171

一般演題（ポスター）

Verification of phenotype of novel atopic dermatitis model IL-33Tg mice

〇野々村 徹、石田 裕紀、西川 英俊、大津 麻未、佐々木 麻衣、山崎 則之
株式会社 新薬リサーチセンター・非臨床研究部

In the development of drug discovery for atopic dermatitis（AD）, chemical-induced, special diet-induced or
Tg mice have been used in experiments using rodents.
In order to overcome AD, it is expected that an animal model that appropriately reproduces the pathologicalcondition of human AD can be created and utilized for pathological analysis and drug discovery research.
IL-33 is known to induce inflammatory cytokines or increase eosinophils by signaling from the nuclear
transcription factor NFkB.
In the clinical picture of AD, it has been reported that IL-33 is highly expressed in human skin.
The model overproduces IL-33 in mice skin, induces inflammatory mediators via signaling from nuclear
transcription factors, and results in Th2-dominant predominance, leading to AD.
In this presentation, introduce phenotypes such as itching behavior and histological examination of IL-33Tg
mice.

Copyright © The Japanese Pharmacological Society2-B-P-172

一般演題（ポスター）

Oral administration of Lactobacillus AZABU isolated from the gut microbiome ofhealthy dogs significantly prevents the development of atopic dermatitis andallergic asthma in mice models.

〇早川 千春1、大平 智春1、金木 真央1、安田 伊武希1、市川 茉南1、竹田 志郎2、内山 淳平3、福山 朋季1
麻布大・獣医・薬理、2麻布大・獣医・食品科学、3岡山大・院医歯薬・病原細菌

1

Regulation of the gut microbiome using probiotics such as
preventive measures for allergy. We originally isolated
guts. We here evaluated the anti-allergic effects of

spp. is recently focused on as one of thesp. (AZABU) from the healthy dog

AZABU using hapten-induced atopic dermatitis (AD) andasthma mouse models. First, daily oral administration of live

AZABU (2×108 CFU/ml) during theexperimental period significantly prevented the development of AD symptoms, skin thickness, and transepidermal water loss in the AD mouse model. Significant decreased number of IgE-positive B cells in auricularlymph node was also observed inregulated by

AZABU treatment group, indicating allergen-specific immunoreaction was

AZABU. Group 2 innate lymphoid cells, which play a pivotal role in non-allergen-specificimmunoreaction, also significantly suppressed by oral administration of live
effects of live

AZABU. Next, the anti-allergic

AZABU was simultaneously demonstrated in the asthma mouse model, including significant

decrease of inflammatory pathological change of lung, and both allergen-specific and non-specificimmunoreaction in hilar lymph node and lung tissue. Our findings suggested that live
allergic properties although the mechanism of action is still being investigated.

Copyright © The Japanese Pharmacological Society

AZABU has anti-

2-B-P-173

一般演題（ポスター）

Examining anti-allergic properties of live and killed Lactbacillus reuteri isolated
from healthy dogs in a mouse model of atopic dermatitis.

〇市川 茉南1、早川 千春1、大平 智春1、金木 真央1、安田 伊武希1、竹田 志郎2、内山 淳平3、福山 朋季1
麻布大・獣医・薬理、2麻布大・獣医・食品科学、3岡山大・院医歯薬・病原細菌

1isolated from healthy dogs as a probiotic supplement for

We are currently developing

canine atopic dermatitis (AD). In this study, we examined the anti-allergic effects of live

and killed

strain mixture (i.e., strains M01, M11, M40, and M41) using a hapten-induced-AD mouse model. First,
when UV-killed(overgrowth) was orally administered to AD mice, cutaneous barrier function

evaluated by the trans epidermal water loss was maintained as normal in thetreatment group,compared with the control group. Immune reactions including lymphocyte proliferation in auricular lymph
nodes and total IgE levels in serum are significantly regulated by treatment of killed. In contrast,

treatment improve neither AD score nor skin thickness. Second, the anti-AD effects were examined(i.e., M01, M11, M40, or M41). The symptoms of AD and immune

using 4 individual strains of live

responses such as lymphocyte proliferation and total IgE levels are significantly reduced by

M11administration compared with the vehicle control and other strains. Currently, the interaction with macrophages
and regulatory T cells is being studied.

Copyright © The Japanese Pharmacological Society2-B-P-174

一般演題（ポスター）

Effects of glycyrrhizic acid and its metabolite on the GIRK channel activity〇陳 以珊、西谷（中村） 友重
和歌山県立医科大・医・薬理学講座

G-protein-gated inwardly rectifying K+ (GIRK) channels control various physiological functions. For example,
GIRK1/2 heterotetramers in the brain regulate neuronal excitability; GIRK1/4 heterotetramers in the heart
regulate heart rate. GIRK channels are potential therapeutic targets for several diseases, such as atrial fibrillation
and addiction. In the present study, we aimed to identify the effect of glycyrrhizic acid (GA), a main ingredientof licorice, and its metabolite glycyrrhetic acid (GRA) on GIRK channel activities. By electrophysiologicalrecordings using

oocytes expressing different GIRK subunits, we observed that GA inhibits the currentof heteromeric GIRK1/2 and GIRK1/4 but slightly activates the current of homomeric GIRK2 and GIRK4. Thissuggests that the inhibitory effect of GA is GIRK1-dependent. Mutation of a GIRK1-specific amino acid residue
in the pore helix, Phe137, to Ser abolishes the inhibition of GIRK current by GA, suggesting that the Phe137
plays important roles in the sensitivity of channels to GA. Unlike GA, GRA activates all GIRK2, GIRK4,
GIRK1/2 and GIRK1/4 channels. Taken together, these data indicate that GA and GRA have distinct actions on
GIRK currents, and would provide clues to elucidate the diverse mechanisms of GIRK channel regulation by
analyzing the difference of these compounds.

Copyright © The Japanese Pharmacological Society2-B-P-175

一般演題（ポスター）

RAGE, which is the receptor for advanced glycation end-products, is involved in
the regulation of KCC2 expression by the Porphyromonas gingivalis LPS
treatment in PC-12 cells.

〇富田 和男1,2、古川 紗圭1,3、五十嵐 健人1,2、田中 康一1,2,4、北中 純一2、北中 順惠4、西山 信好2、野口 和行3、佐藤 友
昭11

鹿児島大・院医歯・歯科薬理、2兵庫医科大・薬・薬理、3鹿児島大・院医歯・歯周病、4兵庫医科大・医・薬理-

KCC2 has been shown to be important for neural maturation. We have shown thatderived lipopolysaccharide (LPS) decreases KCC2 expression. We have also shown that oxytocin,

Kamishoyosan, and Kamikihito restore the KCC2 decrease by

LPS treatment. However, the detailed

molecular mechanism is still unclear. On the other hand, RAGE (receptor for advanced glycation end-products;
AGE) was discovered as a receptor of AGE at first, it has been also reported as a receptor for LPS and oxytocin
now. Therefore, we analyzed the relationship between
Treatment reduced the expression of
LPS. In addition,, the receptor for

LPS and RAGE using PC-12 cells. SiRAGE
LPS, and canceled the KCC2 decrease by

LPS treatment results in the nucleus localization of REST and MECP2 that bind to thetranscriptional regulatory region of, whereas siRAGE treatment inhibits the nuclear localization of REST

and MECP2. Furthermore, Kamishoyosan treatment directly reduced the expression oftreatment increased the expression of, and Kamikihito, which is one of the ligands of RAGE. These results suggest thattargeting RAGE can control the expression of KCC2, which is important for neurological maturation, and that
Kamishoyosan and Kamikihito are potential therapeutic agents for neurological disorders.

Copyright © The Japanese Pharmacological Society2-B-P-176

一般演題（ポスター）

Antiviral activity of curcumin and its analogs selected by artificial intelligencesupported activity prediction system in SARS-CoV-2-infected VeroE6 cells

小松 弘嗣1、田中 剛史1、叶 正成1、池田 健1、松崎 尹雄1、城間 保2、細田 雅人1、安木 真世3,4,5、〇手島 浩慈2
インタープロテイン、2レキオファーマ・研究開発本部、3大阪公立大学・院獣医、4大阪公立大学・アジア健康科学研究1所、5大阪公立大学・大阪国際感染症研究センター

Curcumin has been reported to exert its anti-SARS-CoV-2 activity through multiple mechanisms including
inhibition of spike receptor-binding domain (RBD) to angiotensin-converting enzyme-2 (ACE2) binding. To
identify more potent compounds, we tested curcumin and its analogs for spike RBD-ACE2 binding inhibitory
activity and antiviral activity in SARS-CoV-2-infected cells. An artificial intelligence (AI) -supported activity
prediction system was used to select the compounds, and 116 compounds with a docking score range of -8.7 to-4.3 kcal/mol were selected from 334 curcumin analogs. These compounds were narrowed down to 10compounds, including curcumin, for confirmatory studies. These 10 compounds showed a significant correlation( =0.685, P=0.029) between the IC20 values of spike-RBD-ACE2 binding inhibitory activity and EC50 values ofantiviral activity, indicating that the antiviral activity was mediated by spike RBD-ACE2 binding inhibition.
Based on the assumption that the binding site of curcumin and its analogs is different from that of anti-spike
RBD antibody drugs, it is expected that these compounds through pharmaceutical or pharmacokinetic
modification, or the development of more potent derivatives would contribute to supplementing the antiviral
activity of antibodies against SARS-CoV-2.

Copyright © The Japanese Pharmacological Society2-B-P-177

一般演題（ポスター）

Antimicrobial activities of ginseng saponins isolated from Red Ginseng to nontuberculous mycobacteria〇寒川 訓明1、山口 雄大2、徳留 健太郎1、本間 拓二郎1、松永 慎司1、冨田 修平1
大阪公立大学・医・分子病態薬理学教室、2(国研)国立感染症研究所・細菌第一部

1

Non-tuberculous mycobacterial lung disease (NTM lung disease) is the infection which is caused bymycobacteria, such as

(

),

,

. The prognosis of NTM

lung disease is relative benign than that of TB, but in some cases NTM lung disease is highly resistant tochemotherapy. The incident rate of NTM lung disease is increased and it is estimated that the number of NTM
patients exceeds the number of tuberculosis (TB) patients. Here, we investigated the antimicrobial activity ofginseng saponins from red ginseng (Red Ginseng Extract; RGE) against NTM.
We examined whether RGE had antimicrobial activity against
growth of

MAH104 strain. RGE inhibited theat more than 2.0 mg/mL. To elucidate antimycobacterial mechanism of RGE, weestablished a resistant strain to RGE. RGE-resistant strain grew slower than wild strain. In acid-fast staining,cell wall staining tended to be attenuated in RGE-resistant strain than that in wild type. Protein expression incell wall fraction from RGE-resistant strain showed some differences from that from wild type.
These results suggest that RGE has the antimicrobial activity against

in vitro. The mechanism of theantimicrobial activity is assumed to be through its effect on the cell wall synthesis.

Copyright © The Japanese Pharmacological Society2-B-P-178

一般演題（ポスター）

Effectiveness of Lecture and Practical on Kampo Medicine for Nursing
Undergraduates: The Relationship between Self-Reported Health Status andInterest in Kampo Medicine〇金岡 麻希、野末 明希、児玉 みゆき、内田 倫子、竹山 ゆみ子、木下 由美子、柳田 俊彦
宮崎大・医・看A survey was conducted using a self-administered questionnaire to clarify the relationship between nursing
undergraduates' perceptions of their own health status and their interest in Kampo medicine, and to clarify theirchanges after a lecture and practical. About 85% of the nursing students considered themselves “very healthy”
or “healthy”. Before the lecture, the higher health levels corrected with, lower interest in Kampo medicine.
Conversely, after the lecture, interest in Kampo medicine in the healthy group increased. This suggests that
undergraduate lecture can increase interest in Kampo medicine, especially among healthy nursing students.
Regardless of their health status, the nursing students tended to recognize the need for knowledge of Kampo
medicine before the lecture. This had a particular impact on the students in the healthy group, who accounted
for the largest proportion of students surveyed. Before the lecture, students were aware of the need for Kampo
medicine in nursing education. However, following the lecture, the healthy and poor health groups were moreaware of the need for Kampo medicine.

Copyright © The Japanese Pharmacological Society2-B-P-179

一般演題（ポスター）

Suppressive effects of ergothioneine on Aβ-induced hyperphosphorylation of tau
protein in SH-SY5Y cells〇柴垣 郁弥1、小菅 葉利1、板花 将輝1、加藤 優希1、松本 聡2、中道 範隆1
高崎健康福祉大・薬・分子薬物治療、2エル・エス コーポレーション

1

Ergothioneine (ERGO) is a hydrophilic antioxidant contained in the food and is distributed to the brain after
oral intake, exhibiting a neuroprotective effect. ERGO protected PC12 cells against cellular toxicity induced byamyloid beta (Aβ) and improved impairment of learning and memory ability in mice administered with Aβ.
However, the effects of ERGO on hyperphosphorylation of tau protein is unclear. In the present study, weinvestigated whether ERGO suppresses Aβ-induced hyperphosphorylation of tau protein in humanneuroblastoma SH-SY5Y cells. SH-SY5Y cells were differentiated using culture medium containing 1% or 10%
fetal bovine serum (FBS) with 10 μM retinoic acid. Exposure to Aβ25-35 clearly decreased cellular viability in
SH-SY5Y cells differentiated in the 10% FBS medium, but not 1% FBS medium. Pretreatment with ERGO
protected SH-SY5Y cells against cellular toxicity induced by Aβ25-35 in a dose-dependent manner. Exposure of
SH-SY5Y cells to Aβ25-35 increased expression of phosphorylated tau protein, and the increase was suppressedby pretreatment with ERGO in a dose-dependent manner. These results suggest that ERGO may suppress
hyperphosphorylation of tau protein and alleviate neurotoxicity induced by Aβ.

Copyright © The Japanese Pharmacological Society3-B-P-180

一般演題（ポスター）

Apolipoprotein E-containing lipoproteins protect retinal ganglion cells from
NMDA-induced excitotoxicity with reducing α2-macroglobulin via an LRP1 in
retinal glia

〇林 秀樹、森 みすず、南 厚徳、見世 加南子、高木 教夫
東京薬科大・薬・応用生化

Low-density lipoprotein receptor-related protein 1 (LRP1) is a multifunctional receptor that is abundantly
expressed in the central nervous system. In our previous study, glia-derived apolipoprotein E-containing
lipoproteins (ELPs) protected retinal ganglion cells (RGCs) from glutamate-induced excitotoxicity via an LRP1
in vitro. We have also shown that α2-macroglobulin (a2M) interferes with the protective effect of ELPs. It isreported that a2M is increased in vitreous humor of glaucoma patients. However, the details of its function are
unknown. Here we observed that-methyl-D-aspartate (NMDA)-induced excitotoxicity in retinae of rats

showed RGC degeneration and increased amount of a2M in vitreous humor three days after NMDA injection
into vitreous humor. Then, intravitreal injection of ELPs suppressed RGC degeneration and the increased
amount of a2M. In addition, ELPs decreased a2M mRNA and protein levels in primary cultured retinal glia.
LRP1 siRNA blocked the inhibitory effect of ELPs on a2M expression, and the addition of ELPs enhanced thephosphorylation of STAT3 in retinal glia. Thus, ELPs demonstrate optic nerve protection from excitotoxicity
and also reveal an indirect protective effect by suppressing the a2M expression, which is a neuroprotective
disturbing factor. We hope that a reduction of a2M mediated by the ELP-LRP1-STAT3 pathway might be an
additional protective mechanism against excitotoxicity in the retina.

Copyright © The Japanese Pharmacological Society3-B-P-181

一般演題（ポスター）

Regulations of cysteine uptake and intracellular glutathione levels by purine
derivatives〇松村 暢子、角田 ワッタナポン、青山 晃治
帝京大・医・薬理Purine derivatives such as caffeine and uric acid have neuroprotective activities and reduce the risk ofneurodegenerative diseases such as Alzheimerʻs disease and Parkinsonʻs disease. We have reported that caffeine,uric acid and paraxanthine, a major metabolite of caffeine, promote cysteine uptake in hippocampal slices. We
have also reported that paraxanthine promotes the cysteine uptake and the synthesis of intracellular antioxidant
molecule, glutathione (GSH), in HEK293 cells.
In this study, we examined the effect of uric acid, an end-product of purine metabolism, on cysteine uptake andGSH synthesis in HEK293 cells. Uric acid was treated to HEK293 cells for 30min at concentrations of 0, 10 and
100 μM in cystine-deprived condition, and then cysteine was added for 30 min at concentrations of 100 and
400 µM. High performance liquid chromatography analysis showed that uric acid decreased the extracellular
cysteine levels. Fluorescent GSH detection by CMFDA indicated that uric acid increased intracellular GSH
levels. These results suggest that uric acid promotes cysteine uptake leading to GSH synthesis in HEK293 cells.

Copyright © The Japanese Pharmacological Society3-B-P-182

一般演題（ポスター）

Ghrelin enhances excitability of cerebellar molecular layer interneurons andfacilitates GABAergic transmission onto cerebellar Purkinje cells〇廣野 守俊、中田 正範
和歌山県立医科大・医・生理学第２講座

Ghrelin is an endogenous orexigenic peptide for growth hormone secretagogue receptor type 1a (GHS-R1a).
Ghrelin is produced not only in the stomach but also in the brain. A recent study has reported that rodent
cerebellar Purkinje cells (PCs) express GHS-R1a and its activation facilitates spontaneous firing of PCs.
However, little is known about whether ghrelin alters synaptic transmission onto PCs and modulates firing ofPCs. We examined effects of ghrelin on inhibitory GABAergic transmission using patch clamp recordings
applied to mouse cerebellar slices. We found that bath-application of ghrelin increased the frequency of
spontaneous inhibitory postsynaptic currents (IPSCs) in PCs. The peptide did not affect miniature or
stimulation-evoked IPSCs in PCs. Ghrelin significantly increased the spontaneous firing rate of molecular layerinterneurons (MLIs). The ghrelin-mediated effect on MLIs was blocked by a GHS-R1a antagonist JMV3002
and attenuated by blockers of transient receptor potential canonical type 1 (TRPC1) or KCNQ channels,suggesting that ghrelin signaling in MLIs regulates both TRPC1 and KCNQ channels. These results indicatethat the ghrelin-induced potentiation of spontaneous IPSCs is attributed to the firing facilitation of MLIs by theactivation of GHS-R1a expressed most likely in somatodendritic sites of MLIs. Thus, ghrelin controls firing ofPCs directly and indirectly, and could contribute to the regulation of motor coordination.

Copyright © The Japanese Pharmacological Society3-B-P-183

一般演題（ポスター）

Copyright © The Japanese Pharmacological Society3-B-P-184

一般演題（ポスター）

Effect of 5-aminolevulinic acid on the object recognition test in mice

〇小宮 素良、高橋 純平、古賀 愛理、山田 大輔、斎藤 顕宜
東京理科大・薬・薬理学研究室

An endogenous amino acid 5-aminolevulinic acid (5-ALA) is a precursor of heme in the porphyrin synthesis
pathway. It has been reported that 5-ALA synthase is decreased in the brains of patients with dementia,
suggesting an association between cognitive function and 5-ALA. However, little is known about the role of 5ALA in cognitive function. To address this issue, we evaluated learning and memory performance in the novel
object recognition (NOR) test by using ddY mice (4-6-week-old). 5-ALA hydrochloride or vehicle wasintracerebroventricularly administered 30 min before the training session. On the test day, 48 hours after the
training session, 5-ALA (3 mM) -treated mice showed significantly higher ratio of contact time to novel object
compared to the vehicle-treated mice. This result suggested that 5-ALA produced the enhancement of theobject recognition memory in mice. Next, we examined a memory-related synaptic plasticity, long-term
potentiation (LTP), of field excitatory post-synaptic potential (fEPSP) in area CA1 of the hippocampus. As theresults, 5-ALA (300 mM) perfusion in the hippocampal slices significantly increased the slope of the fEPSP,
suggesting the enhancement of the LTP. Based on these results, we propose that 5-ALA could have theenhancing effects on cognitive function in rodents.

Copyright © The Japanese Pharmacological Society3-B-P-185

一般演題（ポスター）

Neurotensinergic neurons in the lateral hypothalamus are important forphysiological sleep-wake cycle.

〇長沼 史登1、中村 正帆1、ベトリベラン ラマリンガム2、吉川 雄朗3、岡村 信行1
東北医科薬科大・医、2ハーバード大学・医・神経学分野、3東北大・院医・薬理学

1

Neurotensin works as a neuropeptide in the brain. We have reported that experimental stimulation ofneurotensinergic neurons in the lateral hypothalamus (Nts-LH) caused hyperarousal in mice. However, itremains unclear whether Nts-LH is involved in the physiological sleep-wake cycle. Thus, in the present study,
we investigated the effect of Nts-LH selective lesion on physiological sleep-wake cycle. We also measured
neuronal activities of Nts-LH during sleep and wakefulness. First, we performed Nts-LH lesion induced bydiphtheria toxin-A. The Nts-LH lesion caused a fragmentation of wakefulness and decreased total amount of
wakefulness, implying that Nts-LH was critical in the transition and maintenance of wakefulness. Next, wemeasured the neuronal activities of Nts-LH during sleep and wakefulness by using fiber photometory with a Ca2+

sensor, GCaMP7f. The calcium imaging clearly showed that Nts-LH was simultaneously activated at thetransition from sleep to wakefulness, followed by continuous excitation during wakefulness. These dataemphasize that Nts-LH mediate sleep-to-wake transition then their activation promote the following
wakefulness. Taken together, the neuronal activity of Nts-LH maintains wakefulness, which is important forphysiological sleep-wake cycles

Copyright © The Japanese Pharmacological Society3-B-P-186

一般演題（ポスター）

Albumin suppresses hydrogen peroxide- and nimustine-induced cell death via
increasing intracellular GSH levels in human astrocytoma U-251 MG cells.

〇吉岡 靖啓、砂川 和也、田野井 一、山室 晶子、石丸 侑希
摂南大・薬We previously demonstrated that albumin increases intracellular glutathione (GSH) levels in humanastrocytoma U-251 MG cells. GSH has been known to play an important in oxidative stress tolerance andanticancer drug resistance in tumor cells. In this study, we investigated the effect of albumin on hydrogen
peroxide- and nimustine (ACNU)-induced cell death in U-251 MG cells using bovine serum albumin (BSA).
Intracellular GSH levels were determined by the DTNB recycling assay. Cell death of U-251 MG cells was
examined by Hoechst 33342/Propidium iodide staining. Treatment of U-251 MG cells with BSA (0.1-10
mg/mL) increased the levels of intracellular GSH and catalytic subunit of glutamate-cysteine ligase (GCLc), a
rate-limiting enzyme in GSH synthesis, protein in a concentration-dependent manner. In addition,
pretreatment with BSA for 24 h attenuated hydrogen peroxide (0.75 mM)- and ACNU (1 mM)-induced cell
death in a concentration-dependent manner. This cytoprotective effect of BSA was not observed in the presenceof DL-Buthionine-(S,R)-sulfoximine (1 mM), an inhibitor of glutathione synthesis. These results suggest thatalbumin increases intracellular GSH levels in U-251 MG cells by inducing GCLc protein and that albumin
suppresses hydrogen peroxide- and ACNU-induced cell death via increasing intracellular GSH levels.

Copyright © The Japanese Pharmacological Society3-B-P-187

一般演題（ポスター）

Activation of mechanosensitive Piezo1 channel suppresses brown adipocyte
differentiation〇内田 邦敏1、剣持 麻奈斗1、川原崎 聡子2、瀧澤 咲月1、岡村 和彦3、後藤 剛2
1

静岡県立大・食品栄養科学・生体機能学、2京都大・院農・食品生物科学、3福岡歯科大・生体構造

Brown adipocytes cause an energy consumption by heat production and are thought to be a target for theprevention of obesity and related metabolic disorders. Piezo1 is a Ca2+-permeable non-selective cation channel
and activated by mechanical stimuli. While, Piezo1 has been reported to be involved in mechano-sensation in
non-sensory tissues, the expression and role of Piezo1 in brown adipocytes have not been well clarified. In thisstudy, we evaluated a brown adipocytes line from UCP1-mRFP1 transgenic mice and analyzed thiscell. Application of Yoda-1, a Piezo1 agonistsuppressed brown adipocytes differentiation in a dose-dependent
manner. This suppression was significantly recovered by co-application with a Piezo1 antagonist and a
calcineurin inhibitor. In addition, knock-down of Piezo1 impaired Yoda-1-induced suppression of brown
adipocyte differentiation and application of Yoda-1 enhanced the calcineurin activity. These results suggest that
activation of Piezo1 might suppress the differentiation through calcineurin pathway in brown pre-adipocytes.

Copyright © The Japanese Pharmacological Society3-B-P-188

一般演題（ポスター）

KPR-5714, a novel TRPM8 antagonist, improves frequent urination in a modelwith enhanced bladder afferent nerve activity〇渡邉 信次郎1、松澤 亜可根2、小林 淳一1、藤森 芳和1
1

キッセイ薬品工業・研究本部 研究統括部 創薬探索研究所、2キッセイ薬品工業・研究本部 薬理研究所

Transient receptor potential melastatin 8 (TRPM8) has a role in the abnormal sensory transduction of the
bladder and is involved in the pathophysiology of hyperactivity bladder disorders. In the present study, we
examined the effects of KPR-5714, a novel and selective TRPM8 antagonist, on voiding dysfunction induced bybladder afferent hyperactivity via mechanosensitive C-fibers in rats. In cystometry measurements, theintercontraction interval was decreased by intravesical instillation of 10 mM ATP in female rats. KPR-5714
dose-dependently prolonged the shortened the intercontraction interval provoked by ATP. In voiding behavior
measurements, intratesticular injection of 3% acetic acid and water avoidance stress exposure decreased themean voided volume and increased voiding frequency in male rats. KPR-5714 dose-dependently increased themean voided volume and decreased voiding frequency without affecting the total voided volume in these rats.
However, KPR-5714 did not influence the voiding behavior in normal rats. These results suggest that KPR-5714
improves voiding dysfunction by inhibiting the enhanced activity of mechanosensitive bladder C-fibers in rats
with bladder overactivity and does not affect normal voiding behavior.

Copyright © The Japanese Pharmacological Society3-B-P-189

一般演題（ポスター）

Improvement of muscle weakness by nalfurafine hydrochloride (TRK-820), a
kappa opioid receptor agonist〇上野 賢也、森山 正樹、大森 優、上田 寛、内田 将史
東レ・医薬研究所

Cancer cachexia is a progressive functional disorder that cannot be completely reversed with normal nutritional
support. It is a multifactorial syndrome characterized by a persistent loss of skeletal muscle mass (with orwithout fat loss), and anamorelin is the only drug approved for the treatment of cancer cachexia in Japan.
Recently, we found that nalfurafine hydrochloride (our development code TRK-820), which is used as atreatment for pruritus in dialysis and chronic liver diseases refractory to existing therapies, increased food intake
in normal animals, increased body weight and prolonged life in a B16/F10 cell transplantation model mouse,
and improved food intake, body weight gain and muscle weakness in an A549 cell transplanted non-small celllung cancer model mice and may be a potential therapeutic agent for cancer cachexia. . In addition, genetic
analysis of the hypothalamus of A549 cell transplantation model mice revealed that TRK-820 increased peptides
involved in hyperphagia and suppressed dopamine-related genes. These results suggest that TRK-820
suppresses the effects of dopamine in the hypothalamus, causing activation of the hyperphagic system, leading
to increased food intake and consequent weight gain. This presentation will discuss the therapeutic effects of
TRK-820 on cancer cachexia and the details of the mechanisms currently known.

Copyright © The Japanese Pharmacological Society3-B-P-190

一般演題（ポスター）

Effect of nalfurafine hydrochloride (TRK-820), a kappa opioid receptor agonist, inthe treatment of cancer cachexia.

〇大森 優、内田 将史、岩村 智勝、瀬戸 真由美、鈴木 知比古
東レ（株）・医薬研究所

Cancer cachexia is a progressive functional disorder that cannot be completely reversed with normal nutritional
support. It is a multifactorial syndrome characterized by a persistent loss of skeletal muscle mass (with orwithout fat loss), and anamorelin is the only medicine approved for the treatment of cancer cachexia in Japan.
We have previously found that nalfurafine hydrochloride (our development code TRK-820), which is used as atreatment for pruritus refractory to existing therapies for dialysis and chronic liver disease, increased food intake
and body weight and improved muscle weakness in a mouse model of non-small cell lung cancer inoculated with
A549 cells. These results suggest that TRK-820 is a potential therapeutic agent for cancer cachexia. This
presentation will discuss the efficacy of TRK-820 in treating cancer cachexia in animal models and detail themechanisms currently known.

Copyright © The Japanese Pharmacological Society3-B-P-191

一般演題（ポスター）

Suppression of mitochondrial depolarization of neurons by KCC2 inhibitor
VU0463271.

〇原 友凜亜、平嶋 未佳、尾松 果奈、金城 俊彦、宇野 恭介、倉本 展行
摂南大・薬・機能In neurons, massive calcium influx through NMDA receptor causes excitotoxicity by depolarizing mitochondria
by opening mitochondrial permeation transition pore (mPTP). Through mPTP, potassium ions (K+) also passthrough from the cytosol to mitochondrial matrix, and also causing depolarization. We hypothesized that the K+
concentration gradient between the cytosol and the matrix defines how damage the cells. Potassium chloride
cotransporter 2 (KCC2) fundamentally play a role to exclude intracellular chloride ion (Cl-) out in matureneurons by using the driving force of potassium gradient. In this study, we experimentally investigated whether
increasing the intracellular K+ concentration by inhibiting this transporter increases the degree of
mitochondrial depolarization and the degree of cell death. Exposure of VU0463271, an inhibitor of KCC2 toprimary cultured neurons did not change the MTT reducing ability. Contrary to expectations, pre-incubation of
VU0463271 did not affect NMDA-induced neuronal cell death. Moreover, pre-incubation of VU0463271
lowered the degree of mitochondrial depolarization, which was induced by NMDA or valinomycin, amitochondrial uncoupler. It was suggested that the K+ concentration in the cytosol was constantly regulated bybuffering by mitochondria.

Copyright © The Japanese Pharmacological Society3-B-P-192

一般演題（ポスター）

Determination of alteration of intracellular potassium level by using simple
device.

〇岡田 暉己、二股 有貴子、岩本 昂也、金城 俊彦、宇野 恭介、倉本 展行
摂南大・薬・機能We have hypothesized that increased concentration of potassium ion (K+) in the cytoplasm determines thedegree of mitochondrial depolarization and enhancement of cytotoxicity and is an important factor in
determining neuronal cell death. The definition of intracellular ion concentration is possible using the patch
clamp method, while the equipment is expensive. Fluorescence indicators for various ions are still in thedevelopment stage. In this study, we investigated whether it is possible to define intracellular and extracellular K+ concentrations and chase alterations in intracellular K+ concentrations using a simple K+ meter. A calibration
curve of K+ concentration-ppm was prepared with a potassium chloride solution. We could determine the K+
concentration in tissue homogenates from several brain regions, and it was found that the K+ concentration per
tissue weight was almost constant. It was found that when the cell suspension or the medium of adherent cells
was changed to a medium containing no K+, the intracellular K+ concentration decreased. Also in the medium
containing a high concentration of K+, the intracellular K+ concentration increased. The medium of the cellsuspension and adherent cells was changed to a medium containing valinomycin, which is a K+ ionophore, or
VU0463271, which is a K+, Cl- cotransporter inhibitor, however no change in intracellular K+ concentration
was observed. In the next, we will investigate using drugs that more directly change the intracellular K+
concentration (eg, K + channel opener).

Copyright © The Japanese Pharmacological Society3-B-P-193

一般演題（ポスター）

A2BR contribution to form an appropriate sensory network in the cortex via
suppression of mGluR5 a postnatal early development〇宮川 美保1、田中 雅彬1、小泉 修一1,2
山梨大・院医・薬理、2山梨大・山梨GLIAセンター

1

Metabotropic glutamate receptor 5 (mGluR5) is abundant in neonatal astrocytes but decreases with ages, and isalmost absent in the healthy adult brain. Astrocytic mGluR5 is important in formation of excitatory synapses.
We previously showed that A2b receptor (A2BR), being upregulated in astrocytes, is responsible for downregulation of

encoding mGluR5 during postnatal early development. Here, using A2BR knock out mice

(A2BKO), we investigated effects of this loss of inhibitory control over mGluR5 during early development on
adult brain functions. We firstly examinedastrocytes. The

expression level in the healthy adult (7 months old) corticalexpression was twice as high in A2BKO mice as that of control wild-type mice (WT).

Then, we performed two behavioral tests on A2BKO mice (9 to 11 weeks old). In von Frey test on naïve A2BKO
mice, the percentage of maximum pain score of A2B mice was significantly higher than that of the WT mice. In
temperature nociceptive threshold test, A2BKO mice exhibited lower threshold than WT mice. These results
indicated that A2BKO mice are more sensitive to these sensory stimuli than WT mice, suggesting that astrocytic
A2BR would contribute to form an appropriate sensory network in the cortex presumably via suppression ofmGluR5-mediated synaptic remodeling.

Copyright © The Japanese Pharmacological Society3-B-P-194

一般演題（ポスター）

Regulation of directional cell motility in alveolar macrophages by a formin family
protein Fhod1〇三浦 綾子1、實松 史幸2、武谷 立1
1

宮崎大・医・薬理学、2尚絅大学・生活科学・栄養科学科

The actin cytoskeleton functions in various cellular events, including cell motility, morphogenesis, cytokinesis,
and establishment and maintenance of cell polarity. Formin family proteins are structurally characterized by thepresence of the formin homology domains (FH) 1 and 2, and play pivotal roles in actin filament assembly in avariety of cellular processes. Fhod3, a cardiac member of the family, is expressed abundantly in the heart andneurons, while Fhod1 is ubiquitously expressed. Fhod3 plays an essential role in cardiogenesis andneurogenesis. Fhod1 is known to contribute to the assembly of stress fibers, contractile actin structures found incultured non-muscle cells. However, therole of Fhod1 remains poorly understood. Here we show that

Fhod1 is abundantly expressed in the lung. Especially, alveolar macrophages isolated from bronchoalveolar
lavage fluids were strongly stained with anti-Fhod1 antibodies. In Fhod1 KO mice, directional cell motility ofalveolar macrophages was selectively impaired, suggesting the possibility that Fhod1-mediated actin assembly
seems to promote cell polarization during cell migration. We will discuss roles of Fhod1 in the regulation of
directed cell migration in alveolar macrophages.

Copyright © The Japanese Pharmacological Society3-B-P-195

一般演題（ポスター）

Calcineurin B homologous protein 3 facilitates the skeletal muscle development〇古林 創史、野部 浩司
昭和大・薬Calcineurin B homologous protein 3 (CHP3) is a calcium-binding protein and is highly expressed in heart. We
previously found that this protein inhibits cardiomyocyte hypertrophy. However, the function in skeletal muscle
is unknown. In the present study, we examined its role in the differentiation and fusion process of mouseskeletal muscle. CHP3 was barely expressed in adult rectus femoris muscle, but abundantly expressed inneonatal muscle. The expression level of CHP3 was increased in mouse C2C12 myoblasts during thedifferentiation into myocytes. Disruption of the CHP3 gene in the myoblasts with the CRISPR-Cas9 systemdecreased the expression level of myosin heavy chain (MHC), a marker of skeletal muscle differentiation. In
addition, the myotube fusion index (number of myotubes with at least two nuclei per total myotubes) was
decreased. On the other hand, over-expression of CHP3-mCherry increased MHC expression and myotube
fusion. These results suggest that CHP3 regulates skeletal muscle development.

Copyright © The Japanese Pharmacological Society3-B-P-196

一般演題（ポスター）

Prevention of endotoxic death by a nociceptor-derived anti-microbial peptide.

〇近藤 豪1、杉澤 恵利香2、丸山 健太2,3
北海道大・院医・医化学、2大阪大・iFReC、3生理学研

1

Sepsis is a high-mortality disease in which dysregulated production of pro-inflammatory cytokines cause multiorgan defects and lead to endotoxic shock. Despite the extensive use of anti-inflammatory treatments (e.g.
TNF-α antibody or glucocorticoids) in patients undergoing endotoxic shock, its mortality rates remain high at
approx.30%, indicating that the mechanism of endotoxic death is only partially explained by uncontrolledinflammation. In this study, we identified an anti-microbial peptide, Reg3γ, as a protective factor against
endotoxic death. During endotoxemia, nociceptor-derived Reg3γ penetrates into the brain and suppresses the
kynurenine pathway. Endotoxin-administered nociceptor-null mice and nociceptor-specific Reg3γ-deficient
mice exhibit a high mortality rate accompanied by an aberrant level of quinolinic acid and a decreased ATP
production in the brain despite normal peripheral inflammation. Strikingly, the central administration of Reg3γ
protects mice from endotoxic death. These findings provide novel insights into the molecular machinery oftolerance to endotoxic death.

Copyright © The Japanese Pharmacological Society3-B-P-197

一般演題（ポスター）

Research on histone deacetylase 2-selective inhibitors for the treatment ofsynovial sarcoma〇権藤 花奈1、広川 貴次2、吉田 将人1、平尾 巧2、木越 英夫1、竹中 聡3、岩崎 憲治41

筑波大院数理物質、2筑波大医学医療系、3大阪国際がんセ、4筑波大TARA

Synovial sarcoma is a malignant tumor of the soft tissues. Although it is clear that the SS18‒SSX fusion protein
generated by t(X;18)(p11.2;q11.2) drives synovial sarcoma, the underlying mechanism remains unknown,
making drug discovery research difficult. However, it was recently reported that inhibition of histone deacetylase(HDAC) 2 leads to the degradation of SS18-SSX. Unfortunately, HDAC inhibitors usually inhibit multiple
HDACs simultaneously, and there is only one example of an HDAC2-selective inhibitor. Therefore, thedevelopment of inhibitors with high isozyme selectivity would be valuable. In this study, we aimed to design
small-molecule compounds with significant differences in binding affinity between HDAC1 and HDAC2, whichhave very high sequence identity. To achieve this aim, we performed molecular dynamics calculations, organic
synthesis, measurement of dissociation kinetics, and measurement of growth-inhibitory effects on synovial
sarcoma cells. The molecular dynamics calculations of the structures of HDAC1 and HDAC2 without drug
revealed that the pocket volume of the drug binding site is clearly larger in HDAC2, suggesting that selective
inhibitors could feasibly be developed. Based on the pocket volume revealed by MD pocket analysis, we aredesigning drugs with high affinity for HDAC2.

Copyright © The Japanese Pharmacological Society3-B-P-198

一般演題（ポスター）

Elevated lipid peroxidation products that are produced during cell cycle
progression are involved in the execution of ferroptosis〇本間 拓二郎1,2、小林 翔3、藤井 順逸2
1

大阪公立大学・院医・分子病態薬理学、2山形大・院医・生化学分子生物学、3山形大・農Ferroptosis is a newly characterized form of cell death that is caused by the accumulation of intolerable levels of
lipid peroxides in the cell via an iron-mediated Fenton reaction. The resulting lipid peroxides that are generated
in membrane phospholipids cause ferroptotic cell death by disturbing the integrity of the plasma membrane.
Glutathione (GSH), a tripeptide redox molecule that contains a cysteine (Cys) unit in the center, plays a pivotal
role in protection against ferroptosis. Cys is metabolically produced by the transsulfuration pathway inconjunction with methionine (Met) metabolism and may fulfil the requirement in some organs under normal
physiological conditions. In addition to protein synthesis, Met is the precursor for S-adenosylmethionine(SAM), which donates a methyl group to several acceptor molecules such as DNA, RNA, proteins, andphospholipids. Cell growth is arrested in Met-free medium, which characteristically occurs in cancer cells.
Because Met is an essential amino acid, a defect in Met supply could impair the protein synthesis required forcell cycle progression.
Despite the signiﬁcant roles of Met in Cys metabolism and cancer biology, the interplay between Met
metabolism and ferroptosis in cancer cells has not been fully investigated. We had speculated that a combined
deprivation of Met and cystine induces ferroptosis more effectively in vitro. In the current study, however, wefound the Met/cystine double deprivation strongly prevented the execution of ferroptosis under conditions of
intracellular Cys/GSH starvation, which led to the survival of HeLa cells as well as Hepa 1‒6 cells.
Supplementation of SAM resulted in the increased production of peroxidized lipids and induced ferroptosis incells under double deprived conditions. On the other hand, SAM supplementation also increased DNA
methylation and allowed cell cycle progression to resume. These collective results reveal the pivotal roles of lipid
peroxides, the concentrations of which are elevated during cell cycle progression, in ferroptosis execution under
Cys starvation conditions.

Copyright © The Japanese Pharmacological Society3-B-P-199

一般演題（ポスター）

The transcription factor SOX4 regulates phenotypic changes in epithelial
keratinocytes〇長岡 良礼1、武石 幸容1、高橋 千代1,2、武田 佳奈1,2、岡村 和彦3、姚 遠4、本村 香織4、大徳 浩照4、八田 光世11

福岡歯科大・細胞分子生物学講座・分子機能制御学分野、2福岡歯科大・歯・矯正歯科学分野、3福岡歯科大・歯・病態構造学分野、4筑波大・生存ダイナミクス研究センター

SOX4 is a member of the SOX (Sex-determining region Y-related high-mobility group box) family oftranscription factors, and known to associated with the promotion of epithelial cell tumorigenesis, invasion, andmetastasis. However, the role of SOX4 in epithelial keratinocytes remains elusive. In this study, we aim toinvestigate the involvement of SOX4 in the phenotypic changes and functional regulation of a humankeratinocyte cell line (HaCaT).Firstly, we generated a SOX4 overexpressing cell line (Tet on SOX4 HaCaT) in
which SOX4 expression is induced in the presence of doxycycline (Dox). Dox treatment impaired intercellularcontact and altered the cells to exhibit a protruding morphology. Moreover, phalloidin staining revealed anincrease in filopodia (filamentous cell membrane extensions by fibrous actin bundles)-like structures. qRT-PCR
and western blotting revealed that SOX4 decreased the expression of epithelial markers (KRT13, CLDN1) andincreased the expression of mesenchymal markers (Vim, FN1). Because the gene profiles may have beensignificantly altered, we performed comprehensive RNA sequencing analysis. Gene clustering and gene ontology
analyses of differentially expressed genes revealed that Dox treatment of Tet on SOX4 HaCaT increased theexpression of genes related to the cytoskeleton such as actin fiber formation and actin skeletal regulation. Thesefindings indicate that SOX4 induces EMT-like phenotypic and cytoskeletal changes in epithelial keratinocytes.

Copyright © The Japanese Pharmacological Society3-B-P-200

一般演題（ポスター）

Involvement of ceramide kinase in the formation of autophagosomes〇布能 英樹千葉大・院薬・薬効薬理学研究室

Autophagy is the principal intracellular degradation pathway that helps maintain cell homeostasis by degrading
harmful protein aggregates and damaged mitochondria. It is known that abnormalities in autophagy cause
serious diseases such as neurodegenerative diseases and cancer. Thus, autophagy is a fundamental and essential
phenomenon for living organisms, and knowing its mechanism is indispensable for developing disease treatment
and prevention methods. The most significant feature of autophagy is the creation of a new double-membraneorganelle called the autophagosome. However, the involvement of sphingolipids in the formation of
autophagosomes is entirely unknown. Therefore, we investigated the involvement of ceramide kinase (CerK) in
the formation of autophagosomes.
CerK is an enzyme that produces ceramide-1-phosphate (C1P) from the ceramide, “the central moleculein sphingolipid metabolism.” This study found that CerK knockout cells had lower basal autophagy levels thanWT cells. In CerK-KO cells, the treatment with rapamycin or bafilomycin A1 delayed formation of
autophagosomes compared to WT cells. Furthermore, the knockdown of CerK in WT cells resulted in a decreasein the number of autophagosomes. These results indicate that CerK is involved in the formation of
autophagosomes.

Copyright © The Japanese Pharmacological Society3-B-P-201

一般演題（ポスター）

Duloxetine suppresses nitric oxide production induced by lipopolysaccharide in
BV-2 microglia cells〇中谷 善彦1、矢口 真菜美1、荻野 和樹1、野口 理沙子1、山本 直樹1,2,3、天野 託4
1

国際医療福祉大・薬、2国際医療福祉大・基礎医学研究センター、3東京都立大学・神経科学、4栃木県立岡本台病院

It is well known that both selective serotonin and serotonin noradrenaline reuptake inhibitors can improve thesymptoms major depressive disorder to increase the concentration for monoamine in the synaptic cleft based on
the monoamine hypothesis. However, accumulating evidence has indicated that inflammation in the brain may
be a key factor in the pathophysiological mechanisms of developing symptoms of major depressive disorder. In
this study, we focused on whether duloxetine can show the ameliorative effect for inflammatory response
induced by lipopolysaccharide (LPS) in BV-2 microglial cells. Our results indicated that duloxetine decreased
the NO production induced by LPS in a concentration-dependent manner. The increase in the protein
expression level of iNOS by LPS was decreased in a concentration-dependent manner by duloxetine treatment.
Moreover, the increase of the protein expression levels of phosphorylated-IκBα, phosphorylated-Akt and Akt
by LPS were also decreased. Unexpectedly, the protein expression levels of other pro-inflammatory factors suchas COX-2 and phosphorylation ratios for various molecules including IκBα and Akt were not altered by thetreatment of duloxetine. These findings suggest that duloxetine may also act as an anti-inflammatory agent,
which could contribute to its therapeutic effectiveness.

Copyright © The Japanese Pharmacological Society3-B-P-202

一般演題（ポスター）

Lactoferrin enhances neurite outgrowth via TrkA receptor in PC12 cells.

〇水上 乃愛1、長嶋 大地2,3、古川 恵3、東方 優大4、日塔 武彰4、速水 耕介1,2、出雲 信夫2,41

横浜薬科大・薬・機能性物質学研、2横浜薬科大・薬・総合健康メディカル研究セ、3横浜薬科大・薬・薬学教育セ、4横浜

薬科大・薬・薬物治療学研

Lactoferrin (LF) is a multifunctional protein which have the antibacterial and immunomodulatory action in themilk of mammals. Recent studies have been investigated the effect of improvement of cognitive impairment in
the central nerve system. However, the mechanisms underlying the effects of LF on neuron have not yet been
elucidated. The aim of present study investigated the efficacy of LF on the cell outgrowth effect using by PC12
cells, and demonstrated to mechanisms of LF.
PC12 cells were treated with different concentration of LF (100 to 1000 µg/mL) for maximum of 72 h, and weretreated with 50 ng/mL of nerve growth factor (NGF) as a positive control. Digital images of cells were takenwith a phase-contrast inverted microscope with a camera at 24 and 72 hours after treatment of either LF or
NGF. Total ERK and phosphorylated ERK expression levels were analyzed by western blotting, and theinhibitors were used AG879 and PD98059.
The cell outgrowth was significantly increased at 72 hours after NGF treatment. Treatment of 250 µg/mL LF
was significantly increased all of the neurite length, neurite joint, and neurite branching (pass) compared to the
non-treated group. The ratio of total ERK and phosphorylated ERK expression levels was maximized at 5
minutes after treatment of LF, and was persisted up to 10 minutes. These observed activities were inhibited by
AG879 and PD98059.
This present study clarified that the role of LF may enhance the cell outgrowth via activation of phosphorylated
ERK after 5 minutes, and it suggested to undergo nerve outgrowth via the same pathway of NGF.

Copyright © The Japanese Pharmacological Society3-B-P-203

一般演題（ポスター）

Linalool, an essential oil component of lavender, inhibits the activation of
2+
nociceptive transient receptor potential ankyrin 1 (TRPA1) and voltage-gated Cachannels in mouse sensory neurons〇橋本 美穂、高橋 賢次、太田 利男
鳥取大・院・獣医薬理

Linalool, an essential oil component ofis commonly used in fragrances. It is known that linalool has

anxiolytic, sedative, and analgesic actions. However, the mechanism of its analgesic action is not fully clear. Painsignals elicited by the activation of nociceptors on peripheral neurons are transmitted to the central nervous
system. In this study, we focused the effects of linalool on TRP channels and voltage-gated channels, both ofwhich are important for pain signaling via nociceptors in somatosensory neurons. For detection of channel
activity, the intracellular Ca2+ concentration ([Ca2+]i) was measured using a Ca2+ imaging system and membranecurrents were recorded by a whole-cell patch-clamp technique. In mouse dorsal root ganglion neurons linalool
did not affect [Ca2+]i responses to capsaicin and acids, TRP vanilloid 1 (V1) agonists. On the other hand,
linalool suppressed the increases of [Ca2+]i induced by allylisothiocyanate and carvacrol, TRPA1 agonists. In
heterologously expressed channels, linalool suppressed [Ca2+]i responses to TRPA1 agonists but not those to
TRPV1 agonists. Linalool attenuated the [Ca2+]i responses to high concentrations of KCl and voltage-gated Ca2+
currents but only slightly suppressed voltage-gated Na+ currents. These results suggest that linalool exerts ananalgesic action via the suppression of the nociceptive TRPA1 channel and voltage-gated Ca2+ channels.

Copyright © The Japanese Pharmacological Society3-B-P-204

一般演題（ポスター）

TRPV4 activation prevents lipopolysaccharide-induced painful bladder
hypersensitivity in rats.

〇善積 克、渡辺 千寿子、溝口 広一
東北医科薬科大・薬Although most studies have reported that activation of transient receptor potential vanilloid 4 (TRPV4)
contributes to bladder pain and overactive bladder with a cardinal symptom of acute or chronic cystitis, its
involvement in the protective response against bacterial infection in various cultured cells including urothelial
cells has also been reported. In the present study, we investigated the potential benefit of intravesical TRPV4
agonist for painful bladder hypersensitivity produced by a rat model of LPS-induced cystitis and whether its
effects modulate the LPS signal for cytokine release and macrophage phenotype change. The increased bladderpain-related behaviors and voiding frequency caused by LPS were suppressed by concurrent injection into therat bladder of a selective TRPV4 agonist, GSK1016790A. Moreover, the production and secretion ofproinflammatory cytokines (e.g., CXCL1, CXCL10), are suppressed by the presence of GSK1016790A.
Furthermore, TRPV4 activation switched the LPS-stimulated pro-inflammatory M1-type macrophage to theanti-inflammatory M2-type macrophage. These results suggest that that activation of TRPV4 in bladderregulates the proinflammatory response by LPS, and TRPV4 functions may be a promising future therapeutictarget for refractory chronic cystitis.

Copyright © The Japanese Pharmacological Society3-B-P-205

一般演題（ポスター）

Roles of pannexin 1 in the trigeminal ganglion in orofacial mechanical allodynia
following infraorbital nerve injury in rats

〇栗栖 諒子1、三枝 禎2、青野 悠里2、林 良憲3、人見 涼露3、前田 茂4、嶋田 昌彦1、岩田 幸一3、篠田 雅路31

東京医科歯科大・病院・歯科ペインクリニック、2日本大・松戸歯・薬理、3日本大・歯・生理、4東京医科歯科大・院医歯・

歯科麻酔・口腔顔面痛制御学

ATP and glutamate (Glu) are known to be released into the extracellular space through pannexin 1 (Panx1)channels in the cell membrane. Neuronal Panx1 contributes to the development and maintenance of peripheral
inflammation-induced tactile hypersensitivity. In order to study mechanisms of orofacial neuropathic pain, we
analyzed the role of Panx1 in the trigeminal ganglion (TG) of rats with infraorbital nerve injury (IONI). Male
SD rats were used. We measured mechanical head-withdrawal threshold (MHWT) in IONI rats receiving an
intra-TG Panx1 inhibitor or a metabotropic Glu receptor 5 (mGluR5) antagonist and in naive rats receiving
intra-TG mGluR5 agonist administration post-IONI. Glu and Panx1 in TG were measured post-IONI. Panx1,
mGluR5 and Glu synthetase expression in TG were analyzed immunohistochemically and changes in thenumber of mGluR5-P2X3-expressed TG neurons were evaluated. MHWT was decreased after IONI and thisdecrease was reversed by the Panx1 inhibitor and the mGluR5 antagonist. The mGluR5 agonist decreased
MHWT. IONI increased extracellular Glu in TG. Panx1 was expressed in satellite glial cells and TG neurons,
and intra-TG mGluR5 antagonism decreased the number of mGluR5- and P2X3-positive TG neurons postIONI. The present results indicate that IONI facilitates Glu release via Panx1 that activates mGluR5 expressedin nociceptive TG neurons innervating the orofacial region. In turn, P2X3 receptor-expressed TG neurons areenhanced via mGluR5 signaling, resulting in orofacial neuropathic pain.

Copyright © The Japanese Pharmacological Society3-B-P-206

一般演題（ポスター）

Repagermanium (Ge-132) capable of trapping H2S and ATP relieves paclitaxelinduced peripheral neuropathy in mice

〇関口 富美子1、安達 義史1、島田 康弘2、中村 宜司2、川畑 篤史1
近畿大・薬・病態薬理、2浅井ゲルマニウム研究所

1

We have reported that the enhanced activity of Cav3.2 T-type Ca2+ channels caused by endogenous H2S
participates in the paclitaxel (PCT)-induced peripheral neuropathy (PIPN), and that neuron-derived ATP
promotes the PCT-induced macrophage (Mφ) infiltration followed by extracellular release of HMGB1, adamage-associate molecular pattern protein, leading to PIPN development. On the other hand, we have foundthat repagermanium (i.e. Ge-132), once hydrolyzed into 3-(trihydroxygermyl)propanoic acid (THGP), can trap
H2S, in addition to ATP, and inhibit H2S-induced enhancement of Cav3.2 activity and pain. Thus, we evaluatedeffects of THGP on PIPN in mice. In the mice treated with PCT repeatedly, daily i.p. administration of THGP
at 100 mg/kg prevented PIPN development and Mφ accumulation in the sciatic nerves. In Mφ-like RAW264.7
cells, THGP at 10 mM inhibited the cell migration caused by ATP at 0.1 mM, but not extracellular HMGB1
release caused by ATP at 1 mM. Finally, a single i.p. administration of THGP at 100 mg/kg, as well as TTA-A2,
a Cav3 inhibitor, at 1 mg/kg, reversed the established PIPN in mice. These data suggest that THGP prevents
PIPN development by trapping ATP that promotes PCT-induced perineuronal Mφ accumulation and reverses
PIPN most probably by trapping H2S that enhances Cav3.2 activity.

Copyright © The Japanese Pharmacological Society3-B-P-207

一般演題（ポスター）

A TRPV1 antagonist AMG517 alleviates abnormal pain sensitivity and improvessocial deficits in the prenatal valproic acid-induced mouse model of autism〇田原 孟1、今戸 瑛二2,3、川瀬 啓生4、樋口 桃子4、山川 英訓5、小川 公一5、古武 弥一郎1、田熊 一敞6,7、橋本 均
4,7,8,9,10、浅野 智志2、吾郷 由希夫2

広島大・院医・生体機能分子動態、2広島大・院医・細胞分子薬理、3広島大・院医・歯科麻酔、4大阪大・院薬・神経薬

1理、5塩野義製薬、6大阪大・院歯・薬理、7大阪大・院連合小児発達、8大阪大・データビリティフロンティア機構、9大阪大・
先導的学際研究機構、10大阪大・院医・分子医薬

Individuals with autism spectrum disorders (ASDs) show neurobehavioral deficits, characterized by
impairments in social interactions, repetitive behaviors, as well as wide sensory abnormalities. We havepreviously demonstrated that prenatal exposure to valproic acid (VPA) at embryonic day 12.5 (E12.5) causes
autism-like behavioral abnormalities in male mouse offspring. We have also found that prenatal VPA exposure
causes long-lasting mechanical allodynia and spinal microglial activation. In the present study, we aimed toinvestigate the role of the transient receptor potential vanilloid type 1 (TRPV1), a highly validated pain target,
in abnormal pain sensitivity and social interaction deficits in prenatal VPA-treated mice. Behavioral analyses
revealed that prenatal VPA-treated mice exhibited thermal hyperalgesia, mechanical allodynia and increased
capsaicin-induced paw licking. A single administration of AMG517, a TRPV1 antagonist, alleviated both
hyperalgesia and mechanical allodynia. Additionally, AMG517 reversed deficits in social behaviors in prenatal
VPA-treated mice and also increased c-Fos expression, a marker for neuronal activity, in the nucleus accumbens.
These findings suggest that prenatal exposure to VPA might alter TRPV1-mediated signaling and TRPV1
antagonists have a potential to treat certain cases of ASDs.

Copyright © The Japanese Pharmacological Society3-B-P-208

一般演題（ポスター）

Inhibition of histone deacetylase in utero causes spinal microglial activation andmechanical allodynia in mice

〇今戸 瑛二1,2、Sun Samnang3、Huynh Ngoc Bao Tran3、中村 庸輝4、中島 一恵4、森岡 徳光4、木口 倫一5、浅野 智志
1,3、吾郷 由希夫1,3

広島大・院医・細胞分子薬理、2広島大・院医・歯科麻酔、3広島大・歯、4広島大・院医・薬効解析科学、5和歌山県医大・

1

薬・生体機能解析

Valproic acid (VPA) is an anticonvulsant drug that is approved for use in epilepsy and bipolar disorder, but it
also acts as a histone deacetylase (HDAC) inhibitor. We have previously demonstrated that prenatal exposure to
VPA at embryonic day 12.5 (E12.5) causes autism-like behavioral abnormalities in mouse offspring. We havealso found that prenatal VPA exposure causes long-lasting mechanical allodynia and spinal microglial activation.
In the present study, we examined the effects of prenatal exposure to trichostatin A (TSA), a potent and specificinhibitor of HDAC class I/II, on tactile sensitivity and microglial morphology in the spinal cord. Pregnant ICR
mice were intraperitoneally injected with either TSA (1 mg/kg) or vehicle on E12.5. Both male and female
offspring of TSA-treated mothers (defined as TSA-treated mice) showed a significant decrease in withdrawal
threshold in the von Frey test. The numbers of microglia in laminae I-IV and V-VI of the spinal cord dorsal hornin TSA-treated mice were increased compared with those in control mice. Increases in average intensity and cellarea per microglia in laminae I-IV and V-VI were also observed in TSA-treated mice. These findings suggestthat inhibition of HDAC during pregnancy causes mechanical allodynia associated with spinal microglial
activation.

Copyright © The Japanese Pharmacological Society3-B-P-209

一般演題（ポスター）

Disulfiram produces potent anxiolytic-like and antinociceptive effects in rodents〇太田 有紗1、寺島 裕也2、山田 大輔1、中谷 百伽1、山内 つぐみ1、坂田 壮太1,2、松浦 航太1,2、藤塚 亮次1,2、重本 千宙
1、吉岡 寿倫1、松島 鋼治2、斎藤 顕宜11

東京理科大・薬・薬理学研究室、2東京理科大・生命医科学研究所・炎症・免疫難病制御部門

Disulfiram (DSF) is an FDA approved drug for the treatment of alcoholism. The drug acts by inhibiting
aldehyde dehydrogenase, an enzyme essential to alcohol metabolism. However, a recent study has demonstrated
that DSF also potently inhibits the cytoplasmic protein FROUNT, a common regulator of chemokine receptor
CCR2 and CCR5 signaling. Several studies have reported that chemokine receptors are associated with theregulation of emotional and pain behaviors in rodents. Therefore, this study was performed to clarify theseeffects of DSF in rodents. The anxiolytic-like and antinociceptive effects of DSF were investigated using anelevated plus-maze (EPM) test and a formalin test, respectively. DSF significantly increased the amount of timespent in the open arms of the maze without affecting the total open arms entries in the EPM test. Moreover,
DSF decreased the duration of pain-related behaviors in the formalin test. However, no effect in both EPM andformalin tests was seen following administration of the selective aldehyde dehydrogenase inhibitor cyanamide.
These results suggested that DSF produces anxiolytic-like and antinociceptive effects in rodents. We proposethat the inhibitory activity of DSF against FROUNT function provides an effective therapeutic option in anxiety
accompanied with pain.

Copyright © The Japanese Pharmacological Society3-B-P-210

一般演題（ポスター）

Protective role for calcitonin gene-related peptide in atherosclerosis progress in
ApoE knockout mice

〇井上 翔太1、川嶋 心2、橋川 直也1、橋川 成美11

岡山理科大・大学院理学研究科、2岡山理科大・理The calcitonin gene-related peptide (CGRP) is a 37-amino acid peptide, and its various functions such as being
a potent vasodilator, being a cytoprotectant, and inhibiting macrophage infiltration have been reported.
However, its role in atherosclerosis remains unclear. We investigated whether CGRP has a role in atherosclerosis
development in apolipoprotein E‒deficient mice. We have previously reported double-knockout ApoE−/−:CGRP−/−(DKO) mice, increased serum cholesterol levels, atherosclerotic plaque areas, and migration functions inperitoneal macrophages with increase in the level of inflammatory cytokine TNFα. Herein, we investigatedwhether inactivating TNFα improves atherosclerosis in DKO mice. We also investigated whether results similar
with those of DKO could be obtained by administering a humanized monoclonal CGRP antibody,
galcanezumab, to ApoE knockout (ApoE KO) mice. ApoE KO male mice and DKO male mice were fed a highfat diet for 8 weeks, and effects on lesion size and macrophage functions were assessed after 2 weeks. The
etanercept was intraperitoneally administered once a week (5 mg/kg), which caused significant reduction in theaortic root and atherosclerotic plaque area in DKO mice. ApoE KO mice were subcutaneously administered with
galcanezumab once a week (50 mg/kg). Furthermore, TNFα inhibition reduced the macrophage migration.
The galcanezumab increased atherosclerotic lesions similar in DKO mice. These results suggested that CGRP
plays a critical role in inhibitory effect on atherosclerosis progression.

Copyright © The Japanese Pharmacological Society3-B-P-211

一般演題（ポスター）

Chronic volume overload and aldosterone provide arrhythmogenic substrates in
the rat atria via potentiation of proarrhythmic responses to cholinergic activation〇相本 恵美、冨永 理紗、永澤 悦伸、高原 章
東邦大・薬・薬物治療

Cholinergic responses of atria have been known to play an important role in vagally-mediated atrial fibrillation(AF). We investigated influence of chronic volume overload and long-term exposure to higher plasma levels of
aldosterone on electrophysiological responses to cholinergic activation in the rat atria. Rats were divided into
three groups based on receiving sham surgery, aorto-venocaval shunt (AVS) surgery to deliver chronic volume
overload, or AVS plus aldosterone administration (1.0 μg/h, i.p.) using an osmotic minipump. Four weeks laterof the AVS surgery, hypertrophy as well as prolongation of atrial effective refractory period (AERP) wereobserved in the isolated heart. Carbachol at 0.1 and 1.0 μM shortened AERP, reflecting proarrhythmic
response, which was potentiated by AVS and further enhanced by AVS plus aldosterone. Carbachol increased
intra-atrial conduction velocity, reflecting antiarrhythmic response, which was also potentiated by AVS
operation but attenuated by AVS plus aldosterone. These results suggest that chronic volume overload andaldosterone provide arrhythmogenic substrates in the rat atria via potentiation of proarrhythmic responses tocholinergic activation. Attenuation of antiarrhythmic response by AVS plus aldosterone may also modify thegeneration of arrhythmogenic substrates in the rat atria.

Copyright © The Japanese Pharmacological Society3-B-P-212

一般演題（ポスター）

Involvement of the prostanoid EP2 receptor stimuation in the glial cell-mediatedvasodilation in the rat retina in vivo

〇森 麻美、関 陽香、水越 聖、上園 崇、坂本 謙司
帝京大・薬・医薬品作用We have previously found that nitric oxide (NO) derived from neuronal cells acts on glial cells and causes
vasodilation in the rat retina via release of epoxyeicosatrienoic acids (EETs) and prostaglandins. The aim of thisstudy was to identify the prostanoid receptors involved in the NO-induced glial cell-derived vasodilation in the
rat retina.
We used male Wistar rats to examine the effects of intravitreal pretreatment of indomethacin, a cyclooxygenase
inhibitor, PF-04418948, a prostanoid EP2 receptor antagonist, and CAY10441, a prostanoid IP receptor
antagonist, on the changes in the retinal arteriolar diameter induced by intravitreal injection of NOR3, an NO
doner. The retinal arteriolar diameters were measured using the ocular fundus images captured with a highresolution digital camera.

Increase in the retinal arteriolar diameter induced by intravitreal injection of NOR3 was significantly suppressedby intravitreal pretreatment of indomethacin and PF-04418948, but not CAY10441.
These results suggested that activation of arachidonic acid cascade and subsequent stimulation of prostanoidEP2 receptors are involved in the rat retinal vasodilatory responses evoked by NO-induced glial cells stimulation.
Thus, glial cells-derived prostaglandin E2 may play important roles in the retinal vasodilatory mechanisms.

Copyright © The Japanese Pharmacological Society3-B-P-213

一般演題（ポスター）
2+

Actions of Myosin Modulators on Contractility and Ca Transients in Healthy andFailing Human Hearts

Abi-Gerges Najah、Sweat Katrina、Truong Ky、Stafford Alexa、Miron Yannick、Roup Ana、Mai Christina、林 隆志、Page Guy、Ghetti Andre
AnaBios Corporation

Understanding the impact of myosin modulators (myotropes) on human heart biology is key to the successful
development of drugs targeting heart failure (HF) and obstructive hypertrophic cardiomyopathy (OHCM).
Omecamtiv mecarbil (OM; positive myotrope; Ph3 for HFrEF) increased contractility (EC50=0.6µM) in isolated
ventricular myocytes from organ donors, while negative myotropes exerted differential effects: Blebbistatin(BBS), N-benzyl-p-toluene sulphonamide and mavacamten (MAVA; approved for OHCM) decreased
contractility (IC50s 2.6, 16 and 0.2µM, respectively), hydroxy-BBS, (S)-3'-aminoBBS and para-amino-BBS
caused biphasic responses. While none of the myotropes impacted peak amplitude of Ca2+ transients, controls
increased (isoproterenol) and decreased (verapamil) peak Ca2+ amplitude. Moreover, OM generated similar
increases in isometric force of ventricular trabeculae across healthy, HFrEF and HFpEF donor hearts. In
contrast to OM, MAVA generated different force decreases between normal and HFpEF vs. HFrEF trabeculae.
Thus, our data support the use of primary human heart preparations early in development to maximize clinicalsuccess of the most promising novel myotropes through evaluation of mechanism of action, structure-activity
relationship stratification, and differential impact across related disease-states.

Copyright © The Japanese Pharmacological Society3-B-P-214

一般演題（ポスター）

The Curcumin Analogue GO-Y022 Suppresses Pressure Overload-induced
Systolic Dysfunction〇平子 裕太1、砂川 陽一1,2,3、清水 果奈1,2、船本 雅文1,2、刀坂 泰史1,2,3、浜辺 俊英1,2,3、柴田 浩之4、小見山 麻紀1、長谷川 浩二1,2、森本 達也1,2,3
静岡県立大・薬・分子病態学分野、2国立病院機構京都医療センター ・展開医療研究部、3静岡県立総合病院・臨床

1

研究部、4秋田大・医学系研究科・臨床腫瘍学講座

We previously found that a natural p300 HAT inhibitor, curcumin (CUR), can inhibit
cardiomyocyte hypertrophy and the development of heart failure. We focused on a CUR analog, GO-

Y022, which shows stronger anti-cancer activity than CUR. The purpose of this study was to determine whether
GO-Y022 inhibits p300-HAT activity and can be used as a therapeutic agent for heart failure.
HAT assay using recombinant p300-HAT domain showed that GO-Y022
inhibited p300-HAT activity as well as CUR. Primary cultured cardiomyocytes prepared from neonatal rats weretreated with GO-Y022 or CUR and then stimulated with phenylephrine (PE) for 48 hours. One µM of GO-Y022
suppressed PE-induced histone H3K9 acetylation, hypertrophic response gene transcription, and cardiomyocyte
hypertrophy to the same extent as 10 µM of CUR. C57BL/6j male mice were subjected to transverse aortic
constriction (TAC) or sham operation. The TAC mice were randomly assigned to five groups: Vehicle, CUR at 1
or 50 mg/kg, or GO-Y022 at 0.2 or 1 mg/kg. After 8 weeks daily oral treatment, echocardiographic analysis
showed that 1 mg/kg of GO-Y022 and 50 mg/kg of CUR improved a TAC-induced increase in left ventricularposterior wall thickness and a decrease in fractional shortening.
】These results indicate that GO-Y022 strongly inhibits both PE-induced hypertrophic responses
and pressure overload-induced development of heart failure. These findings suggest that GO-Y022 may be anovel candidate agent for heart failure therapy.

Copyright © The Japanese Pharmacological Society3-B-P-215

一般演題（ポスター）+

Inhibition of transient outward K current: potential prevention of reentrant
arrhythmia developments in long QT syndromes〇津元 国親1、島本 貴生2、青地 悠馬2、九田 裕一1、谷田 守1、姫野 友紀子2、天野 晃2、倉田 康孝11

金沢医科大・医、2立命館大・生命科学部・生命情報学

Excessive prolongation of cardiac action potential duration (APD) is a risk factor for lethal ventriculararrhythmias. Previously, we have elucidated the underlying mechanisms for the development of early
afterdepolarization (EAD)-mediated premature ventricular complexes (PVCs), leading to the occurrence ofreentrant ventricular tachycardia in congenital or acquired long QT syndromes. However, a method to prevent
EAD-mediated PVC developments has not yet been established. This study aims to theoretically determine theeffect of inhibition of the transient outward K+ channel current ( to) on the development of EAD-mediated
PVCs. Our previous study (doi:10.1254/jpssuppl.95.0_3-P-226) investigated the relationship between EAD and
PVC initiation using a 6 × 6 cm myocardial sheet model consisting of 360,000 human ventricular myocyte
model units (Kurata et al., Biophys J, 2005). In the present study, we examined the effect of to inhibition on
PVC initiation under PVC onset conditions by performing simulations of excitation propagation. Intriguingly,
only 10% inhibition of to prevented the development of reentrant arrhythmias evoked by EAD-mediated PVCs.
Inhibition of to by 30% or more completely suppressed EAD developments, resulting in the prevention of PVC
initiation. Based on our results, to inhibitions may prevent EAD development-mediated reentrant arrhythmias
in the long QT syndrome.

Copyright © The Japanese Pharmacological Society3-B-P-216

一般演題（ポスター）

Comparison of voluntary exercise and renin angiotensin inhibitor administration
in a mouse model of dilated cardiomyopathy〇杉原 匡美1、柿木 亮2,5、村山 尚3、三井田 孝1,6、櫻井 隆3、森本 幸生4、呉林 なごみ3
1

順天堂大・医・臨床検査医学、2順天堂大・スポーツ健康科学部、3順天堂大・医・薬理学、4国際医療福祉大・福岡保健

医療学、5城西国際大・経営情報学部、6順天堂大・医療科学・臨床検査

Dilated cardiomyopathy (DCM) is one of major causes of heart failure (HF). Although exercise is regarded as
one of therapies for HF, the effects of exercise on patients with DCM have not been established. A knock-in
mouse model of human inherited DCM, TNNT2 ΔK210, shows similar characteristics to DCM patients. We
have showed that one of angiotensin receptor blockers (ARB) is effective on cardiac function and electrical
remodeling and found that voluntary exercise also improves cardiac function in homozygote model mice (DCM
mice). In this study, we examined combination therapy in DCM mice. The DCM mice showed enlarged heart
and frequent sudden death with t1/2 of ~70 days. Male DCM mice were divided into 4 groups based on theadministration of ARB and voluntary exercise: without drug or exercise (control), oral administration of ARB(ARB), daily exercise (ex), and combined ARB and exercise (comb). The ex and comb groups started wheel
running at 1 month of age. Cardiac function was measured with echocardiography at 2.5 months of age. Aftersacrifice, weights of body, heart, lung, and lower extremity muscles were measured. Gene expressions of HFand arrhythmia-related genes in myocardium were quantified by qPCR analysis. On echocardiography, theejection fraction was signiﬁcantly improved in only comb-group (Cont: 21.7±6.1％(n=6), ARB: 26.9±5.1％(n=6), ex: 27.2±4.5％(n=5), comb: 31.9±3.6％(n=6)). On the other hand, the heart weight/body weight ratio
decreased in the ARB and the comb groups. Our results indicate a synergistic effect between ARB and voluntary
exercise to cardiac function in DCM.

Copyright © The Japanese Pharmacological Society3-B-P-217

一般演題（ポスター）

Comparative Study of Transcriptome in the Hearts Isolated from Mice, Rats, andHumans〇岡本 洋介1、岡田 大瑚2、小林 大礎1、尾野 恭一1、石井 邦明31

秋田大学大学院、2京都大・院医・ゲノム医学センター、3山形大・医・薬理学講座

The heart is a critical organ for maintaining life in mammals and has long been one of the most important
targets of scientific research, and the basic molecular mechanisms of heart beat appear to have already been
established. However, few studies have focused on species differences, and challenges remain in studying genesthat have universal functions across species and genes that determine species differences. Here, we analyzed
transcriptome data from mouse, rat, and human atria, ventricles, and sinus node (SA) and calculated andcompared specificity measure (SPM) values that account for species differences among the three cardiac
regions. SA has the largest species differences and we searched for a gene, which by our criteria was SHOX2; theSPM value for SHOX2 was prominently high across species. Similarly, SPM values identified 3 atrial-specificmarkers, 11 ventricular-specific markers, and 17 SA-specific markers. Ontology analysis identified 70 cardiac
region- and species-specific ontologies. These results suggest that reanalysis of existing data by calculating SPM
values may identify novel tissue-specific and species-dependent gene expression. This study demonstrates thatSHOX2 is an SA-specific transcription factor, a novel cardiac region marker, and that species-dependent
ontology is important. No COI.

Copyright © The Japanese Pharmacological Society3-B-P-218

一般演題（ポスター）

Mode of prolongation in the early (J-Tpeak) and late (Tpeak-Tend) repolarization can
predict onset pattern and probability of spontaneous termination of following
torsade de pointes attack〇神林 隆一1、後藤 愛1、中瀬古（泉） 寛子1、武井 義則1、松本 明郎2、杉山 篤1,2
1

東邦大・医・薬理、2東邦大・医・加齢薬理IKr inhibitors can prolong the repolarization, and may develop trigger (premature ventricular
contraction: PVC) and substrate (spatial dispersion of repolarization) for the onset of torsade de pointes(TdP). We investigated how the mode of preceding changes in the early (J-Tpeak) and late (Tpeak-Tend)
repolarization represent the characteristics of TdP.
Well-known torsadogenic drug hydroxychloroquine in doses of 1, 3 and 10 mg/kg/10 min was
intravenously administered to the chronic atrioventricular block dogs under the monitoring of Holter
electrocardiogram (n=4 for each dose).
While the low or middle dose of hydroxychloroquine did not induce TdP, the high dose induced TdP ineach animal at 10.0 (#1), 8.7 (#2), 9.3 (#3) and 11.3 (#4) min after the start of administration. The R on T-type
PVCs were frequently induced in the animal #1, #2 and #3 before the onset of TdP, which was not observed in
the animal #4. The TdP spontaneously terminated in the animal #1, while it degenerated into ventricularfibrillation in the others. The changes of J-Tpeak just before the onset of PVC (#1, #2, and #3) or TdP (#4) were+37, +36, +23 and +1 ms, whereas those of Tpeak-Tend were +12, +20, +22 and +89 ms, respectively.
The prolongation of J-Tpeak may trigger R on T-type PVCs possibly through Ca2+ overload, whereas
the prolongation of Tpeak-Tend will develop the substrate for maintaining spiral reentry.

Copyright © The Japanese Pharmacological Society3-B-P-219

一般演題（ポスター）

Involvement of regional specificity of glial cells in neuronal alpha-synuclein
expression and neurodegeneration in parkinsonian models〇宮崎 育子1、菊岡 亮1、磯岡 奈未1、十川 千春2、十川 紀夫3、北村 佳久4、浅沼 幹人1
1

岡山大・院医歯薬・脳神経機構、2広島工業大・生命・生体医工、3松本歯科大・院歯・遺伝子工学・分子創薬、4就実大・薬・薬物治療Exposure to pesticides, such as rotenone or paraquat, increases the risk of Parkinson's disease (PD). Various
studies revealed the link between pesticide toxicity and cellular pathology in PD such as alpha-synuclein (alphaSyn) aggregation and neuronal death. Recently, we demonstrated region-specific astrocyte-microglia interaction
promoted rotenone-induced non-cell-autonomous dopaminergic neurodegeneration. In this study, we examined
effects of regional difference in glial cells on neuronal alpha-Syn expression and neurotoxicity. We prepared
mesencephalic neuronal culture and glial cell culture (astrocyte+microglia) from mesencephalon or striatum of
SD rats embryos (E15). Treatment with conditioned media from mesencephalic, but not striatal, glial cell
culture upregulated alpha-Syn expression in mesencephalic neurons, which were dopaminergic, serotonergic
and GABAergic neurons. Conditioned media from rotenone-exposed mesencephalic, but not striatal, glial cells
increased alpha-Syn expression in dopaminergic and serotonergic neurons; however, neuronal damage wasobserved only in the dopaminergic neurons. These results suggest that regional specificity of glial cells could
contribute to neuronal alpha-Syn expression, and that some factors in addition to alpha-Syn accumulation are
involved in dopaminergic neurodegeneration.

Copyright © The Japanese Pharmacological Society3-B-P-220

一般演題（ポスター）

Mechanisms underlying mutant astrocyte-mediated demyelination in Alexander
disease〇久保田 友人1,2、繫冨 英治1,2、齋藤 光象1,2、篠崎 陽一1,2、小林 憲司1,2、田中 謙二3、池中 一裕4、大野 伸彦5、小泉 修
一1,2
1

山梨大・院医・薬理、2山梨大・院医・GLIAセンター、3慶應大・医・先端医脳科学、4生理学研・分子神経生理、5自治医大・医・解剖

Alexander disease (AxD), a rare neurodegenerative disease, is caused by the mutation of
et al.,

gene (Brenner, 2001) encoding glial fibrillary acidic protein (GFAP) which is enriched in astrocytes. Thus,

AxD is a primary astrocyte disease. AxD patients mainly show severe neurological symptoms such as
psychomotor developmental delay, motor deficits etc. with white matter disorders (leukodystrophy). However,molecular pathogenesis that leads from astrocytic mutations to leukodystrophy and further to these clinicalmanifestations are not well understood. Here, we show that AxD astrocytes directly cause demyelination in
corpus callosum (CC) using AxD model mice carrying human mutant GFAP with R239H (Tanaka et al.,,2007). First, we found by immunohistochemical analysis that myelinated area stained with either myelin basic
protein or proteolipid protein in CC was markedly decreased in AxD compared to wild-type (WT) mice.
Second, the number of astrocytes was increased in CC of AxD compared to WT. In addition, AxD astrocyteswith Rosenthal fiber, a hallmark of AxD, were highly accumulated in demyelinated area. Such astrocytic
accumulation was not observed in CC of WT. These spatial correlation of AxD astrocytes with demyelination of
CC would suggest that pathological astrocytes may be directly involved in local demyelination in CC. Third, we
performed RNAseq analysis of AxD astrocytes and found that Galectin-3 and lipocalin 2 (Lcn2) were the top 10
most up-regulated genes (Saito & Shigetomi et al.,
, 2017) and Lcn2 (Wan et al.,

, 2018). Interestingly, both Galectin-3 (Morizawa et al,, 2022) could positively control astrocytic

phagocytosis. Together, all these findings suggest that AxD astrocytes may cause demyelination by acquiring
their abnormal phagocytic ability in CC, thereby leading to leukodystrophy and various neurological symptoms.

Copyright © The Japanese Pharmacological Society3-B-P-221

一般演題（ポスター）

Astrocytes in the PVN are involved in corticotropin-releasing factor-induced
sympathetic activation in rats

〇山口 奈緒子、岡田 尚志郎
愛知医大・医・薬理Corticotropin-releasing factor (CRF) plays a key role in stress responses in the brain. CRF activates not only the
hypothalamic-pituitary-adrenal axis but also the sympathetic nervous system. We previously reported that both
central administration of CRF and exposure to acute restraint stress (RS) increase plasma catecholamine levels
and activate presympathetic neurons in the paraventricular hypothalamic nucleus (PVN). However, it is unclear
whether glial cells in brain regions including the PVN are involved in CRF- or stress-induced sympathetic
activation. In this study, we examined roles of astrocytes in the CRF-induced elevation of plasmas catecholaminelevels (noradrenaline and adrenaline) using fluorocitrate which blocks glial metabolic function.
Intracerebroventricular pretreatment of fluorocitrate suppressed the CRF-induced elevation of plasma levels of
noradrenaline and adrenaline. On the other hand, microinjection of fluorocitrate into the PVN suppressed the
CRF-induced elevation of adrenaline, but not of noradrenaline. Furthermore, we tried to examine effects ofinhibition of astrocytes on RS-induced sympathetic activation. Our results suggest that astrocytic function in thebrain is important for the CRF-induced sympathetic activation and the effects might be brain region-dependent.

Copyright © The Japanese Pharmacological Society3-B-P-222

一般演題（ポスター）

Long-term stimulation of α7 nicotinic acetylcholine receptor selectively
suppresses thrombin-polarized M1 microglia〇大西 正俊、町田 葵、井上 敦子
福山大・薬The effect of nicotine on impaired M1 and protective M2 microglial polarization was investigated using BV-2
cell line. Alpha 7 nicotinic acetylcholine receptor (α7 nAchR) expression was transiently increased by nicotine
treatment and then it was gradually decreased for 14 days. Treatment with nicotine for 14 days slightly polarizednaïve M0 microglia to CCL1-positive M2b and inducible NO synthase (iNOS)-positive M2d subtypes.
Conversely, arginase1-positive M2c microglia was decreased. On the other hand, exposure of thrombin
recruited iNOS- and interleukin-1β-double positive M1 microglia. Long-term treatment with nicotine
significantly decreased thrombin-induced iNOS mRNA and oppositely showed the tendency to increase thearginase1 mRNA level. From the effect of nicotine on M0 microglia, it is unlikely that nicotine has shifted from
iNOS-positive M1 to arginase1-positive M2 subtype. Thus, the selective cell death of M1 microglia waspresumed. The mitogen-activated protein kinase (MAPK)s pathway has previously shown to be involved in thesurvival of thrombin-activated microglia. Treatment with nicotine for 14 days suppressed thrombinphosphorylated p38 MAPK through the α7 nAchR. These results suggest that long-term stimulation of α7nAchR causes suppression of thrombin-activated p38 MAPK followed by possible apoptosis in M1 microglia.

Copyright © The Japanese Pharmacological Society3-B-P-223

一般演題（ポスター）

Suppressive effects of glatiramer acetate on methamphetamine-induced
preference with microglial osteopontin upregulation in mice

〇泉尾 直孝、玉谷 隆典、中島 卓海、高橋 晃、浅野 昂志、新田 淳美
富山大・薬・薬物治療[Background] Methamphetamine (METH) is a widely distributed addictive drug in the world. Since there is noeffective treatment for METH addiction, novel therapeutic agents are required. Previously, we reported thatmicroinjection of osteopontin (OPN), immune-modulating extracellular matrix protein, to the nucleus
accumbens reduced METH preference behavior (2017;7(1):13084) The study has offered a possibility

that upregulation of osteopontin in the brain is a therapeutic strategy for the drug addiction. Here, we focusedon the glatiramer acetate (GA), which is reported to increase OPN in the brain（
2017;67:163-180） and examined the eﬀects of GA on METH preference. [Methods] Mice (C57BL/6J, 8-9
weeks, male) were received subcutaneous GA (0.1 mg) administration for 14 days before and during behavioral
experiments. METH (1 mg/kg) was treated to these mice during behavioral experiments. OPN-positive cells
were detected in the flowcytometry experiments. [Results] GA suppressed METH-induced conditioned place
preference, not hyperlocomotion. Number of OPN-positive cells increased by GA administration, and most cells
were microglia. [Discussion] GA ameliorated preference behavior induced by METH, which was possibly
mediated by microglial OPN upregulation. GA could be a candidate of a therapeutic agent for drug addiction.

Copyright © The Japanese Pharmacological Society3-B-P-224

一般演題（ポスター）

Evaluation of oligodendrocyte precursor cell properties in a 3-dimension (3D)
culture system〇中野 静香1,2,3、植田 尭子1、松永 行子2,3、村松 里衣子1
精神神経セ・神経研・神経薬理、2東大院・工・バイオ、3東大生研

1

Axons in the central nervous system (CNS) are wrapped with myelin formed by oligodendrocytes. CNS damage
causes demyelination which leads to neurological dysfunction. Remyelination is a regenerative process, which ismediated by oligodendrocyte precursor cells (OPCs) development. Mechanisms underlying OPCs development
have been extremely investigated with in vitro models using 2-dimension (2D) culture systems. Although 3D
culture easily offers model mimicking physical and mechanical properties of cell adhesion or microenvironment
in vivo, little is known about the difference of OPCs properties between in 2D and in 3D culture. Here, wecharacterized the phenotypic difference of OPCs cultured in between 2D and a collagen-based 3D culture
system. OPCs isolated from murine mixed glial culture were embedded in type I collagen gel (3D, final
concentration: 2.4 mg/mL) or plated on glass substrate (2D), respectively. Immunocytochemical analysis
showed a significantly low level in the percentage of Ki67+ cells in Olig2+ cells in 3D compared with 2D,
suggesting that OPCs have less proliferative capacity in 3D. When differentiation was induced bytriiodothyronine, a low level of the expression of myelin basic protein was observed in 3D compared to 2D,
suggesting that OPCs have less differentiation capacity in 3D. Collagen gel can easily control the stiffness bychange in concentration, therefore, these results provide crucial insights to understand OPCs response in 3D
culture, which contributes to develop a 3D model mimicking pathological conditions.

Copyright © The Japanese Pharmacological Society3-B-P-225

一般演題（ポスター）

Postischemic voluntary running exercise promotes the survival of astrocytesborn after cerebral ischemia and alters astrocytic gene expression〇山口 菜摘、澤野 俊憲、中谷 仁、田中 秀和
立命館大・院生命科学

Cerebral ischemia causes neuronal damage and functional impairment. Dendritic spine dynamics is involved infunctional recovery. We have previously reported that voluntary running exercise after focal cerebral ischemia
ameliorates the ischemia-induced dendritic spine loss in the peri-infarct motor cortex layer 5. Environmental
change surrounding neurons affects the neuronal morphologic plasticity. The aim of this study is to reveal theeffect of postischemic exercise on the glial phenotypes. We found that voluntary running exercise promoted thesurvival of astrocytes born after middle cerebral artery occlusion (MCAO) until postoperative day 15.
Transcriptome analysis of astrocytes detected 10 upregulated genes and 70 downregulated genes in exercisegroup compared with non-exercise group. Gene ontology analysis showed that the downregulated genes wererelated to apoptosis and neuronal morphology. Exercise tended to decrease the expression of

known

to reduce dendritic spines in astrocytes after MCAO. Our data suggest that voluntary running exercise after
cerebral ischemia alters astrocytic gene expression, which contributes to the amelioration of the ischemiainduced dendritic spine loss.

Copyright © The Japanese Pharmacological Society3-B-P-226

一般演題（ポスター）

Role of astrocytes in nicotine-induced motor excitement symptoms〇國澤 直史、加藤 将貴、小田 明奈、白川 美波、坂口 茉鈴、清水 佐紀、大野 行弘
大阪医科薬科大・薬・薬品作用解析学

Nicotine elicits motor excitement symptoms such as tremor and convulsive seizures. We performed behavioral
studies in combination with the brain mapping analysis of glial fibrillary acidic protein (GFAP), a marker ofastrocytic activation, to clarify the role of astrocytes in nicotine-induced tremor and convulsive seizures.
Intraperitoneal (i.p.) injection of nicotine induced straub tail and tremor at 0.3 or 1 mg/kg, and convulsive
seizures at 3 mg/kg in mice. Immunohistochemical examination revealed that nicotine region-specificallyelevated GFAP expression in the piriform cortex (PirC) at 1 mg/kg (i.p.), and CA3 area and dentate gyrus (DG)of hippocampus at 0.3-3 mg/kg (i.p.). We next evaluated the effect of fluorocitrate (FC), an astrocyte inhibitor,
on nicotine-induced tremor and convulsive seizures. Intracerebroventricular injection of FC (1 nmol) reduced
GFAP expression in PirC, basolateral amygdala, CA2, CA3 and DG. Whereas FC did not affect the generationof nicotine tremor, it caused a significant decrease in seizure intensity and a decreased incidence of seizures.
These results suggest that astrocytic activation by nicotine in PirC, CA3 or DG is involved in nicotine-induced
convulsive seizures.

Copyright © The Japanese Pharmacological Society3-B-P-227

一般演題（ポスター）

Microglial replacement ameliorates Rosenthal fiber accumulation in Alexander
disease, a primary astrocyte disease.

〇小林 憲司1,2、繁冨 英治1,2、檀上 洋右1,2、Parajuli Bijay1,2、久保田 友人1,2、齋藤 光象1,2、田中 謙二3、池中 一裕4、小泉 修一1,2
山梨大・院医・薬理学、2山梨大・院医・GLIAセンター、3慶應義塾大・医・先端研・脳科学、4生理学研・分子神経生理

1

Alexander disease (AxD) is a rare neurodegenerative disease caused by the pathogenic variants of, anastrocyte-specific intermediate filament. This resulted in the formation of aberrant inclusions within astrocytes
called Rosenthal Fibers (RFs), one of the main pathological hallmarks. In addition, activation of microglia (MG)is also observed in the AxD brain. We previously reported a beneficial role of MG using the AxD model mice,overexpressing mutant human GFAP. Here, we enhanced this beneficial function of MG by replacing MG using
PLX5622 (PLX), a CSF-1R antagonist. Turning PLX ON and OFF causes MG removal and repopulation
respectively (Rep), which allows us to replace old MG with repopulated MG (Rp-MG) in the AxD mice.
Interestingly, the RFs were significantly reduced by Rep, indicating that Rep rescued the AxD pathology. We
further analyzed the bulk RNA-sequencing data and found that lysosomal and phagocytic pathways were highly
enhanced by Rep. Additional histological analysis revealed that Rp-MG strongly expressed CD68 and engulfed
RFs, suggesting Rep could increase MG phagocytic capacity to remove RFs. Overall, the MG replacement could
restore the AxD pathology by increasing their phagocytic capacity against RFs.

Copyright © The Japanese Pharmacological Society3-B-P-228

一般演題（ポスター）

Establishment of a screening method for the developmental neurotoxicity ofchemicals using the image analysis of the dynamics of neurons, astrocytes, andmicroglia in zebrafish〇弓削 瑞葵1、若井 恵里1、小岩 純子1、白水 崇1、駒田 致和2、西村 有平1
三重大・院医・統合薬理学、2近畿大・理工・発生生物学

1

A wide variety of substances that may affect the development of the central nervous system (CNS) in humanexist in environment. Evaluating how chemical teratogens adversely affect the developing brain may provide
important insights not only for the management of industrial chemicals but also for the prevention andtreatment of CNS diseases. However, a system for efficiently and comprehensively evaluating the developmental
neurotoxicity (DNT) of chemicals has not yet been sufficiently established. Here, we try to establish theassessment of DNT analyzing the dynamics of neurons, astrocytes, and microglia in developing zebrafish CNS,
with particular focus on the property of microglia reflecting neuroinflammation. We generated a transgenic
zebrafish line expressing different fluorescent proteins; Cerulean for neurons, mCherry for astrocytes, andmVenus for microglia. Subsequently, we exposed various neurotoxicants to zebrafish embryo and performed invivo fluorescence imaging. We are trying to quantitatively measure the total amount, distribution, andmorphology of neurons, astrocytes, and microglia in the larval zebrafish brain by imaging analysis and detect
DNT comparing with already reported findings.

Copyright © The Japanese Pharmacological Society3-B-P-329

一般演題（ポスター）

Miyako Bidens Pilosa extract regulates microglia polarization in a mouse modelof amyotrophic lateral sclerosis at advanced-stage〇鶴田 こむぎ、設樂 尊人、高橋 愛、宮岸 寛子、小菅 康弘
日本大・薬

is generally used as ethnomedicine and functional food worldwide. We have reported previouslythat Miyako Island Bidens pilosa extract (MBP) suppresses glial activation and prolongs the life span in the
SOD1G93A transgenic mouse model of Amyotrophic Lateral Sclerosis (G93A). However, the therapeuticmechanisms of MBP remain unclear. In the present study, we investigated the microglia activation andpolarization profile in the spinal cord of G93A. Real-time PCR revealed that oral administration of MBP at 2
g/kg/day for 7 days was inhibited the induction of M1-microglial markers (CD11c, IFN-γR) and inflammatory
cytokines (TNF-α, IL-1β, IL-6). In contrast, the increased expression of M2-microglial markers (IL-13R,
Ym1) and anti-inflammatory cytokines (TGF-β, IL-10) was not affected. Moreover, we determined the effect of
MBP on cell proliferation using BV2 microglia cell line. Exposure of BV2 cells to MBP, resulted in a reduction of
MTT reduction activity without increase the number of ethidium homodimer-1 stained dead cells. We also
found that MBP suppressed the LPS-induced pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) production.
These results suggest that MBP regulates neuroinflammation through suppressing the microglia polarizing into
M1 type.

Copyright © The Japanese Pharmacological Society3-B-P-229

一般演題（ポスター）

GPR143 expressed in the dorsal striatum positively regulates behavioral
responsiveness to dopamine D2 receptor agonist quinpirole.

〇大瀧 百々代、増川 太輝、北村 慧、井上 美優、田近 伶、荒井 柾美、五嶋 良郎
横浜市立大・院医・分子薬理神経生物学

L-3,4-dihydroxyphenylalanine (L-DOPA) by itself has been believed to be an inert amino acid that exerts actionand effectiveness in Parkinsonʻs disease through its conversion to dopamine by aromatic L-amino aciddecarboxylase. We propose that L-DOPA is a neurotransmitter. We recently identified a G-protein coupled
receptor 143 (GPR143), as a receptor for L-DOPA. GPR143 is distributed in some specific brain regions
including striatum and substantia nigra in which dopamine receptors are highly expressed. To investigate therole of GPR143 in dopaminergic transmission, we examined the effect of quinpirole, a dopamine D2 receptor(D2R) agonist, on mouse behavior. Quinpirole (0.03 mg/kg, i.p.) decreased locomotor activity in wild-type(WT) mice. We found that this behavioral response to quinpirole was attenuated in Gpr143 gene-deficient(Gpr143-/y) mice when compared to WT mice. In contrast, SKF81297 (2.5 mg/kg, i.p.), a D1R agonist,
increased locomotor activity in Gpr143-/y mice to a similar extent to that in WT mice. To validate GPR143
function after its depletion, we performed rescue experiment of GPR143 using adeno-associated virus encoding
GPR143-P2A-enhanced green fluorescent protein (EGFP) or EGFP in Gpr143-/y mice. The decreased
responsiveness to quinpirole in Gpr143-/y mice was rescued by re-expression of GPR143 in the dorsal striatum.
These results indicate that GPR143 positively regulates D2R function in the dorsal striatum.

Copyright © The Japanese Pharmacological Society3-B-P-230

一般演題（ポスター）

Characteristics of α-synuclein uptake at the blood-brain barrier〇道具 伸也1、横谷 みき1、高田 芙友子1、岩尾 卓朗1、松本 純一2、佐野 和憲3
福岡大・薬・応用薬剤、2福岡大・薬・生物薬剤、3福岡大・薬・生体機能制御

1

Parkinson disease (PD) is characterized by widespread distribution of aggregated α-synuclein (α-Syn) protein
in inclusions known as Lewy bodies. α-Syn is secreted from neurons and transferred to neighboring cells. This
cell-to-cell transmission is thought to underlie the progress of PD. In addition, it is known that blood α-Synlevels in patients with PD are elevated and blood-borne α-Syn can penetrate into the brain across the bloodbrain barrier (BBB). Therefore, another explanation for elevated brain levels of α-Syn is the increasedtransport of α-Syn from blood to brain across the BBB. Here, we investigated how monomeric α-Syn is taken
up by brain endothelial cells which constitute the BBB. We assessed uptake of α-Syn by brain endothelial cells
as cell/medium ratio using primary cultures of rat brain endothelial cells (RBECs). Increasing concentrations of
α-Syn resulted in the increased cellular accumulation of α-Syn in RBECs. The cell/medium ratio did not show
significant changes with the increased concentration of extracellular α-Syn ranging from 0.05 to 10 μg/mL,
suggesting that α-Syn uptake by RBECs is independent of a saturable transport system. However, the α-Syn
uptake by RBECs showed a temperature-dependent manner, suggesting that α-Syn uptake by RBECs is
mediated by an energy-dependent transport system. This uptake was inhibited by mannan. These results
suggest that α-Syn transport at the BBB would be mediated by a mannan-sensitive transport system including
mannose receptor-mediated transcytosis.

Copyright © The Japanese Pharmacological Society3-B-P-231

一般演題（ポスター）

Role of hyperpolarization activated cyclic nucleotide-gated (HCN) channel 1 inmodulating seizure susceptibility to pilocarpine〇石﨑 悠斗1、清水 佐紀1、毎田 愛莉1、森山 芽衣1、佐納 匠1、宮越 樹里1、庫本 高志2、大野 行弘1
1

大阪医薬大・薬・薬品作用解析学、2東京農業大・農・動物栄養

Hyperpolarization activated cyclic nucleotide-gated (HCN) channel 1 is abundantly expressed in cerebral
cortex, hippocampus, and cerebellum, regulating spontaneous rhythm and neural oscillation. Previous study
reported that human mutations in thegene are associated with early infantile epileptic encephalopathy

and genetic generalized epilepsy (Brain, 141, 3160-3178, 2018), however, the functional mechanism underlying
seizure generation is still unknown. In this study, we examined the effects of HCN1 channel inhibitor (ZD7288)
and thegene deletion on pilocarpine-induced seizure to elucidate the role of HCN1 channel inmodulating seizure susceptibility. Treatment of mice with ZD7288 significantly increased the incidence of statusepilepticus and the frequency of seizures as compared with control mice.

-KO rats showed a higher seizure

susceptibility to pilocarpine than control (F344) rats. Thus, pilocarpine elicited status epilepticus in all-

KO rats tested, but in none of control rats. In addition, we performed the immunohistochemical analysis of cFos expression following pilocarpine-induced seizure.

-KO rats showed a significantly higher c-Fos

expression than control rats, notably in the cerebral cortex and amygdala. These results suggest that HCN1
channels play an important role in modulating the susceptibility to epileptic seizures, implying that dysfunctionof HCN1 channels cause neuronal hyperactivation of the cerebral cortex and amygdala.

Copyright © The Japanese Pharmacological Society3-B-P-232

一般演題（ポスター）

p-Hydroxyamphetamine induced prepulse inhibition disruptions is modulated byserotonergic and adrenergic receptors〇小野木 弘志1、中川西 修2、根本 亙2、三反崎 聖3、丹野 孝一2、只野 武4
東北福祉大・健康科学・保健看護、2東北医科薬科大・薬・薬理、3高崎健康福祉大・薬・分子神経科学、4金沢大・院医

1

薬保健・環境生態医・公衆衛生

Amphetamine metabolite -hydroxyamphetamine ( -OHA) has been shown to have many pharmacologicaleffects, including causing psychostimulant-induced behaviors such as hyperlocomotion and head-twitch
response. We have previously reported that -OHA induces prepulse inhibition (PPI) disruptions in mice, and
the PPI disruptions are involved in the dopaminergic system. In this study, we investigated the involvement of
the serotonin (5-HT) and noradrenaline (NA) neurotransmission in -OHA-induced PPI disruptions in mice.
As a result, -OHA-induced PPI disruptions were attenuated by pretreatment with ketanserin (a 5-HT2A/2C
receptor antagonist), MDL100,907 (a selective 5-HT2A receptor antagonist), 5,7-dihydroxytryptamine (a 5-HT
neurotoxin), -chlorophenylalanine (a 5-HT synthesis inhibitor) and prazosin (a selective α1 receptor
antagonist), but were not attenuated by pretreatment with DSP-4 (a NA neurotoxin), fuzaric acid (a NAsynthesis inhibitor). These results suggest that the 5-HT neurotransmission is strongly involved in -OHAinduced PPI disruptions, with ancillary involved of α1 receptors.

Copyright © The Japanese Pharmacological Society3-B-P-233

一般演題（ポスター）

Neuroprotective effects of leukotriene receptor blockers on the secretory
phospholipase A2-induced neuronal apoptosis〇矢上 達郎、山本 泰弘、高馬 宏美
姫路獨協大・薬Neurological diseases

brain ischemia are associated with mammalian secretory phospholipase A2 (sPLA2).

The group IB sPLA2 (sPLA2-IB) induced neuronal cell death via apoptosis, which were accompanied withchromatin condensation and DNA fragmentation. Previously, we had established the sPLA2-IB-induced
neuronal apoptosis as themodel for cerebral ischemia. Prior to neuronal apoptosis, sPLA2-IB generates

leukotrienes (LTs) via 5-lipoxygenase (LOX), which are increased in the ischemic brain. Recently, we reported
that LOX inhibitors prevented neurons from the toxicity of sPLA2-IB, suggesting an involvement of LTs to theneurotoxicity of sPLA2-IB. In the present study, we evaluated protective effects of receptor antagonists for LTB4
(LY293111) and cysteinyl LTs (ONO-1078) in the primary culture of rat cortical neurons. The two LT receptor
antagonists suppressed the neurotoxicity of sPLA2-IB in a concentration-dependent manner. sPLA2-IB shrankneuronal cell bodies and shortened neurites Both LY293111 and ONO-1078 ameliorated morphologicaldegenerations such as shrank neuronal cell bodies and shortened neurites in the sPLA2-IB-treated neurons.
Furthermore, these LT receptor blockers prevented neurons from the sPLA2-IB-condensed chromatin andfragmented DNA, suggesting an involvement of LTs in the sPLA2-IB-induced neuronal apoptosis.

Copyright © The Japanese Pharmacological Society3-B-P-234

一般演題（ポスター）

Purinergic receptor-dependent induction of long-term potentiation of inhibitory
synaptic transmission in the rat insular cortex〇小助川 聖史1、山本 清文1、小林 真之11

日本大・歯・薬理学講座、2日本大・歯・歯科矯正学講座

Adenosine triphosphate (ATP) is released from glial cells in the central nervous system, and induces plasticchanges in synaptic transmission. Despite the recent progress in terms of the roles of purinergic receptors in
cerebrocortical excitatory synaptic transmission, their contribution to inhibitory synaptic transmission has been
unknown. Setting the ultimate goal to clarify the role of ATP in neuroplastic changes in inhibitory synaptictransmission in the cerebral cortex, this study aimed to elucidate the effects of ATP and α,β-methylene ATP(αβmATP), a selective agonist of P2X receptors (P2XR), on inhibitory synaptic transmission in the insularcortex (IC). We performed multiple whole-cell patch-clamp recordings from pyramidal cells (PYR) in IC of the
VGAT-Venus transgenic rats. Administration of αβmATP (100 μM) increased the amplitude of unitary IPSC(uIPSC). Interestingly, αβmATP increased the amplitude of miniature IPSCs (mIPSCs), which sustained evenafter washout of αβmATP. The P2XR-dependent potentiation was blocked by intracellular application of an
NMDA receptor (NMDAR) antagonist, MK801. These results suggest that the activation of the P2XR-NMDAR
pathway induces long-term potentiation of IPSCs in the IC.

Copyright © The Japanese Pharmacological Society3-B-P-235

一般演題（ポスター）

Brain cannabinoid CB1 receptors suppress centrally administered bombesininduced facilitation of the rat micturition〇清水 孝洋1、鄒 瑣1、山本 雅樹2、清水 翔吾1、清水 信貴3、東 洋一郎1、吉村 直樹4、齊藤 源顕1
高知大・医・薬理、2高知大・医・小児思春期医学、3高知大・医・骨盤機能セ、4ピッツバーグ大・医・泌尿器科学1

To clarify roles of brain cannabinoid CB1 receptors in regulation of the micturition, we examined the effects
of intracerebroventricularly (icv) administered rimonabant (Rimo, a CB1 antagonist) and JZL195 (JZL, an
inhibitor of enzymes related to degradation of endogenous CB receptor ligands) on icv administered bombesin(BB, a stress-related neuropeptide)-induced facilitation of the micturition in urethane-anesthetized (0.8 g/kg,ip) male Wistar rats. A catheter was inserted into the bladder to perform cystometry. Three hours after thesurgery, JZL (10 or 30 nmol/rat) or Rimo (100 or 300 nmol/rat) was icv administered 30 min before BB
administration (0.01 or 0.03 nmol/rat, icv). BB (0.03 nmol) shortened the intercontraction interval (ICI), an
indicator of micturition frequency, while JZL significantly suppressed the BB-induced response. BB at a lower
dose (0.01 nmol) showed no significant effect on ICI, while the BB significantly shortened ICI after
pretreatment with Rimo. In addition, JZL-induced suppression of the BB (0.03 nmol)-induced response wasabolished in the presence of Rimo. These results indicate that brain CB1 receptors suppress the centrally
administered BB-induced facilitation of the rat micturition.

Copyright © The Japanese Pharmacological Society3-B-P-236

一般演題（ポスター）

Construction of the ELISA assay to quantify Semaphorin 3A in the adult brain〇古川 涼音1、小林 瑞季1、林 克儀2、中村 史雄3,4、櫻井 隆5、五嶋 良郎3、山下 直也1,3,5
1

神奈川工科大・応用バイオ、2カイオム・バイオサイエンス、3横浜市立大・医、4東京女子医科大・医、5順天堂大・医Extracellular soluble signals that control several aspects of neuronal development are known to play a criticalrole in maintaining neuronal function and homeostasis in the mature nervous system. Abnormal expression
and/or secretion of these molecules are therefore thought to be associated with the onset of various types ofneurological disorders. It has been reported that the expression of Semaphorin 3A (Sema3A), a secreted type of
repulsive axon guidance molecule, is impaired in several neurodegenerative disorders. However, due to the lackof a reliable Sema3A antibody, our knowledge about Sema3A expression in the adult brain is still limited. Here
we report the identification of a pair of Sema3A monoclonal antibodies for the sandwich ELISA assay using theAutonomously Diversifying Library system. Both Sema3A monoclonal antibodies recognize the Sema domain ofhuman Sema3A and can measure recombinant Sema3A in the range of 0-100 pM by ELISA. The specificity ofthis assay was confirmed by using the embryonic brains fromdeficient mice as a negative control.

Moreover, this assay could measure Sema3A concentration in Tris buffered saline-and SDS-soluble lysate
obtained from the adult mice brains. These data suggested that our ELISA assay is a reliable tool for thevalidation of Sema3A as a biomarker in neurodegenerative disorders.

Copyright © The Japanese Pharmacological Society3-B-P-237

一般演題（ポスター）

The effect of neurotrophin-3 overexpression in the hippocampus〇笠倉 奈々美、村田 優花、染谷 僚太、田端 遼、瀬木（西田） 恵里
東京理科大・院先進工学・生命システム工学

Neurotrophin-3 (NT-3) is a type of neurotrophic factor expressed in the hippocampus. It has been reported that
NT-3 was upregulated by stress and corticosterone administration in the hippocampus of adult mice. Since
stress suppresses neurogenesis in the hippocampus, we examined whether increasing NT-3 expression
suppresses neurogenesis. We also investigated the effects of increased NT-3 expression on mature neuron
function and stress-induced behavior. We generated NT-3 overexpressing mice in the hippocampus byadministering adeno-associated virus carrying NT-3 gene. Expression of NT-3 in the hippocampus was higher
more than 35-fold higher than in the control group. After 4weeks of NT-3 carrying virus administration, thenumber of proliferating cells and immature neurons in the hippocampal dentate gyrus was decreased in the NT-3 overexpression group. On the other hand, calbindin expression was increased in mature hippocampal
neurons. In behavioral experiments, locomotor activity and anxiety-like behavior tended to be increased in theNT-3 overexpression group. These results suggest that high-dose NT-3 may be involved in the suppression ofstress-induced neurogenesis and anxiety-like behavior.

Copyright © The Japanese Pharmacological Society3-B-P-238

一般演題（ポスター）

Low serum oxytocin levels and autism spectrum disorder-like behaviors in micelacking the cannabinoid CB1 receptors〇縄田 陽子1、安作 美香1、西奥 剛1、山口 拓2
長崎国際大・薬・薬理、2長崎国際大・薬・薬物治療

1

Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disability that demonstrates impaired
social interactions, social communication deficits, and restrictive/repetitive behaviors. It is reported thatchildren with ASD and some animal models of ASD show the abnormality of endocannabinoid (eCB) systems.
To determine the causal role of the eCB systems in the ASD, we have investigated the relationship between theeCB system and ASD-like symptoms, using the cannabinoid CB1 receptor knockout (CB1KO) mice. We foundthat male CB1KO mice demonstrated reduced sociability (3-chambered social approach task) and elevatedrepetitive grooming behaviors (hole-board test). Moreover, CB1KO mice showed resistance to change a learned
pattern of behavior (reversal learning task using T-maze). On the other hands, the serum oxytocin, reported aslower levels in autistic children, significantly decreased in CB1KO mice. These findings suggest that CB1KO
mice show abnormal behavioral phenotypes and endocrine system including social deficits, repetitive behaviors,
cognitive inflexibility and low serum oxytocin levels, which have face and construct validity as an animal modelfor ASD. Therefore, the CB1KO mice will be a valuable tool for the exploration of pathological mechanisms anddevelopment of novel therapeutics in the ASD.

Copyright © The Japanese Pharmacological Society3-B-P-331

一般演題（ポスター）

Study of the molecular mechanism of autophagy activation via nicotine
receptors in a cellular model for Parkinson‘s disease〇滝沢 進之佑、藤牧 綾香、大内 一輝、栗田 尚佳、保住 功、位田 雅俊
岐阜薬科大・薬・薬物治療学研究室[Background] Neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), andamyotrophic lateral sclerosis (ALS) have increasingly been realized to have common cellular and molecularmechanisms including protein aggregation and inclusion body formation. In the present study, to examine
whether the activation of a7 nicotinic receptor has neuroprotective effect against PD-associated mutant asynuclein-induced aggregation and neurotoxicity in a cellular model, we used nicotine and PNU 282987, an a7
nicotinic receptor agonist.
[Methods] To investigate the effect of nicotine and PNU 282987 against mutant a-synuclein aggregates, weevaluated the protein level of a-synuclein in a cellular model of PD.
[Results] We succeeded in generating a new α-Syn stably expressing cell line using a piggyBac transposon
system. Using this system, we investigated the neuroprotective effect of the nicotine and PNU 282987 on asynuclein toxicity. We found that PNU 282987 provided significant protection against a-synuclein-related
neurotoxicity. These results suggest that nicotine and PNU 282987 reduced intracellular aggregatetheactivation of autophagy.
[Conclusions] Nicotine as well as PNU 282987 against a-synuclein-induced aggregation
autophagy are an efficient source of new treatment and prevention for synucleinopathies.

Copyright © The Japanese Pharmacological Society

the activation of

3-B-P-239

一般演題（ポスター）

Docosahexaenoic acid (DHA) selectively inhibits prostanoid TP receptormediated contractions of guinea pig tracheal smooth muscle〇小原 圭将、稲葉 理花子、川北 美礼、De Dios Regadera Montserrat、植竹 智美、村田 梓、西岡 菜々子、黒木 孝
太、追川 俊哉、吉岡 健人、田中 芳夫
東邦大・薬・薬理Docosahexaenoic acid (DHA) is an n-3 polyunsaturated fatty acid abundant in fish oil. Chronic administrationof DHA has been reported to improve asthma due to its anti-inflammatory effects. However, the immediate
effects of DHA on isolated tracheal smooth muscle (TSM) contractility have not been studied. In this study, weinvestigated the potential inhibitory effects of DHA on the guinea pig TSM contractions. DHA (3 × 10−5 M)significantly inhibited TSM contractions induced by U46619 (a thromboxane A2 (TXA2) mimetic, 10−8 M) andprostaglandin F2α (PGF2α, 5 × 10−7 M). The TSM contractions induced by U46619 (10−8 M) and PGF2α (5 ×
10−7 M) were significantly inhibited by SQ 29,548 (a prostanoid TP receptor antagonist, 10−6 M). DHA (4 ×
10−5 M/6 × 10−5 M) shifted the concentration-response curves (CRCs) for U46619 and PGF2α to the right in aconcentration-dependent manner. However, the slope of the regression line in the Schild plot of DHA
U46619/PGF2α was larger than unity. In contrast, DHA (4 × 10−5 M) did not significantly affect the CRCs foracetylcholine, histamine, and leukotriene D4. These findings indicate that DHA selectively inhibit prostanoid
TP receptor-mediated TSM contractions. DHA may have preventive and ameliorative effects on asthma attacks
associated with TSM hyper-contraction caused by TXA2 and PGF2α.

Copyright © The Japanese Pharmacological Society3-B-P-240

一般演題（ポスター）

The ROCK signaling regulates the pulmonary vascular permeability via
maintenance of lung homeostasis.

〇赤嶺 孝祐、寺林 健、佐々木 隆子、石崎 敏理
大分大・医・薬理Rho-associated kinase (ROCK), including the ROCK1 and ROCK2 isoforms, is a protein kinase involved insignaling from Rho to actin cytoskeleton. Many studies on ROCK functions in cultured cells have been reported.
However, functions of ROCK in vivo remain incompletely understood, because targeted disruption of theROCK1 and ROCK2 genes leads to embryonic death. In this study, using a tamoxifen-induced Cre-loxP system,
we deleted ROCK1 and ROCK2 in adult mice (ROCK double conditional knockout mice: ROCK DcKO) and
explore the in vivo functions of ROCK.
ROCK DcKO mice exhibited poor survival and were observed pulmonary leakage of blood cells after tamoxifen
injection. To confirm the increase of vascular permeability in ROCK DcKO, we carried out vascular permeability
assay using Evans blue dye. As a result, massive leakage of Evans blue dye was observed in the lung tissue ofROCK DcKO compared with that of control mice. In addition, the number of neutrophils was increased inbroncoalveolar lavage fluid (BALF) in ROCK DcKO mice. In the lung tissue of ROCK DcKO mice, the intensityof phalloidin staining and VE-cadherin staining was obviously reduced. These results indicated that pulmonary
vascular permeabilities were increased in ROCK DcKO mice. Furthermore, we found that deletion of ROCKs
led to induce the proinflammatory cytokine gene expression through NF-κB and AP-1 by microarray analysis
and qPCR.
Thus, our study suggest ROCK might be involved in maintenance of lung homeostasis though actin cytoskeleton
rearrangement.

Copyright © The Japanese Pharmacological Society3-B-P-241

一般演題（ポスター）

The effects of dasatinib on corticosteroid insensitive airway inflammation inmice induced by tobacco smoke〇西本 裕樹1、安藤 大稀1、入江 孝祐1、開沼 郁美1、片山 侑紀1、佐藤 しおり1、鈴木 智大1、原田 真衣1、吉田 翼1、木
村 元気1、Ito Kazuhiro2、木澤 靖夫1
日本大・薬・機能形態学、2NHLI, Imperial College London, United Kingdom

1

Chronic obstructive pulmonary disease (COPD) is characterized by corticosteroid insensitive airway
inflammation. Tobacco smoke (TS) induces oxidative stress and activation of Src, which cause airway
inflammation. Recently, we reported that dasatinib (DAS), a Src inhibitor, suppressed airway inflammation inmice induced by lipopolysaccharide. Then, in this study, we determined the effects of DAS alone and the
combination with quercetin, an antioxidant agent, on TS induced airway inflammation.
A/J mice were exposed to TS for 11 days, followed by intranasal treatments with DAS, quercetin andfluticasone propionate (FP) twice daily for 3 days. Bronchoalveolar lavage fluid (BALF) was collected one day
after the last drug treatment, and the numbers of inflammatory cells in BALF were measured byhemocytometers and a flow cytometer.
TS induced the signiﬁcant increases in the numbers of neutrophils and macrophages in BALF, and FP had noeffect on these increases. DAS improved the corticosteroid insensitive airway inflammation in mice induced by
TS. In addition, the combination of DAS with quercetin showed further suppressive effects on airway
inflammation in mice than DAS alone. These results suggested that the combination of Src inhibitors withantioxidant agents may be a novel therapeutic strategy for the treatment of COPD.

Copyright © The Japanese Pharmacological Society3-B-P-242

一般演題（ポスター）

RAMP1 signaling attenuates acute lung injury by inhibiting cytokine production
and neutrophil recruitment〇山下 敦1,2、伊藤 義也1、松田 弘美2、長田 真由子1、田邉 美奈1、古江 明子1、細野 加奈子1、畑中 公1、辻川 和丈3、
馬嶋 正隆1,4、岡本 浩嗣2、天野 英樹11

北里大・院医療・分子薬理、2北里大・医・麻酔科、3大阪大・院薬・細胞生理、4神奈川工科大・健康医療

Neuroimmune interactions have emerged as critical regulators of inflammation. Neuropeptide calcitonin generelated peptide (CGRP) regulates cytokine production in immune cells through signaling for CGRP receptor,
receptor activity-modifying protein 1 (RAMP1). Here, we examined the role of RAMP1 signaling in LPSinduced lung injury. Acute lung injury was induced by intratracheal injection of LPS in wild-type mice (WT)and RAMP1 knockout mice (RAMP1 KO). Compared with WT, RAMP1 KO exhibited decreases in survival rateand increases in lung injury score, and total protein concentrations and pro-inflammatory mediators including
IL-6 and CXCL2 in BALF at 72 h after LPS administration. In WT, CGRP and RAMP1 levels in the lung wereincreased after LPS administration, and RAMP1 was expressed in alveolar macrophages. After LPS
administration, the numbers of alveolar macrophages in both types of mice were diminished, and reached nadir
at 6h, and restored to half levels of pre-values in WT thereafter, but remained low in RAMP1 KO until up to 72
h. By contrast, the numbers of neutrophils were increasing with time after LPS administration in two genotypes,
and those in RAMP1 KO were larger than WT at 72 h. These results suggested that RAMP1 signaling
attenuated LPS-induced acute lung injury by inhibiting cytokine production, neutrophil accumulation, andpulmonary vascular permeability.

Copyright © The Japanese Pharmacological Society3-B-P-243

一般演題（ポスター）

Anti-allergic rhinitis activity of tea tree (Melaleuca alternifolia) essential oil inmice sensitized and challenged with Japanese cedar pollen〇牧野 春香1、山下 道生2、安藤 祐介3、笠井 菜穂子1、田中 淑媛1、星野 楓月1、松尾 香寿美1、山下 恵梨華1、山田 萌恵1、吉田 夏子1、竹ノ谷 文子2、渡辺 知恵3、酒井 寛泰4、塩田 清二5、千葉 義彦1
1

星薬科大・薬・分子生物学、2星薬科大・薬・運動科学、3城西大・薬・臨床病理学、4星薬科大・薬・生体分子薬理学、5湘

南医療大学・薬・解剖生理学

Effects of tea tree (TT) essential oil on allergic rhinitis (AR) induced by Japanese cedar pollen (JCP) were
investigated. Mice were sensitized by. injections with JCP+alum on days 0, 7 and 14. From day 21, thesensitized mice were challenged by intranasal (also treated with TT (.) administrations of JCP for 4 consecutive days. Animals were.) 30 min before each JCP challenge. Frequency of sneezing was counted for 20 min

after each nasal challenge. On days 18 and 25, the histamine (Hist,.)-induced sneezing was also counted. In

mice that were sensitized and repeatedly challenged with JCP, both serum levels of IgG and IgE specific for Cry
J1, a major allergen of JCP, were significantly increased: the TT treatments did not affect the increase in Cry J1specific antibodies. In the JCP-sensitized mice, JCP challenge caused a significant increase in sneezing,
indicating that nasal allergic response was induced. The. application of Hist also caused an increase insneezing. The Hist-induced sneezing was further increased significantly on day 25, indicating that nasal
hyperresponsiveness (NHR) had occurred after the repeated JCP challenges. Both the nasal allergic response
and NHR induced by JCP were inhibited by the pretreatments with TT. Thus, TT might be useful for thetreatment of AR induced by JCP.

Copyright © The Japanese Pharmacological Society3-B-P-244

一般演題（ポスター）

Inhibition of Japanese cedar pollen-induced nasal allergic response by lemon
grass (Cymbopogon citratus) essential oil in mice

〇松尾 香寿美1、山下 道生2、安藤 祐介3、笠井 菜穂子1、田中 淑媛1、星野 楓月1、牧野 春香1、山下 恵梨華1、山田 萌恵1、吉田 夏子1、竹ノ谷 文子2、渡辺 知恵3、酒井 寛泰4、塩田 清二5、千葉 義彦1
1

星薬科大・薬・分子生物学、2星薬科大・薬・運動科学、3城西大・薬・臨床病理学、4星薬科大・薬・生体分子薬理学、5湘

南医療大学・薬・解剖生理学

Current study determined effects of lemon grass (LG) essential oil on allergic rhinitis (AR) induced by Japanesecedar pollen (JCP). Male ICR mice were sensitized by. injections with JCP+alum on days 0, 7 and 14. From

day 21, the sensitized mice were challenged by intranasal (Animals were also treated with LG (.) administrations of JCP for 4 consecutive days.

.) 30 min before each JCP challenge. Frequency of sneezing was counted

for 20 min after each nasal challenge. Sneezing induced by histamine (Hist,.) was also counted on days 18

and 25. In the JCP sensitized and repeatedly challenged mice, both serum levels of IgG and IgE specific for Cry
J1, a major allergen of JCP, were significantly increased. The. JCP challenge to sensitized mice caused asignificant increase in sneezing, indicating an induction of nasal allergic response. An increase in sneezing was
also induced by the. application of Hist. The Hist-induced sneezing was further increased significantly on

day 25, indicating an induction of nasal hyperresponsiveness (NHR) after the repeated JCP challenges. Both the
JCP-induced nasal allergic response and NHR were inhibited by the pretreatments with LG. It is thus possiblethat aromatherapy using LG is effective for the treatment of Japanese cedar pollinosis.

Copyright © The Japanese Pharmacological Society3-B-P-312

一般演題（ポスター）

Identification of betuletol from Brazilian Green Propolis that suppresses IL-33
gene expression in Swiss3T3 cells〇水口 博之1、Shaha Aurpita2,3、Islam Rezwanul2,4、北村 嘉章5、武田 憲昭5、福井 裕行1,6
大阪大谷大・薬、2徳島大・院医歯薬・分子情報薬理学、3The Hormel Inst., Univ. Minnesota、4Dept. Biomed. Sci.,

1

Charles E. Schmidt Coll. Med., Florida Atlantic Univ.、5徳島大・院医歯薬・耳鼻咽喉科学、6錦秀会

IL-33 is involved in the pathogenesis of chronic inflammations through the induction of Th2 cytokines andeosinophils. We showed that IL-33 gene expression level was correlated with the blood eosinophils number ofpatients with pollinosis. It suggests that suppression of IL-33 gene expression could ameliorate IL-33-induced
eosinophilic chronic inflammation. It was reported that Brazilian Green Propolis (BGP) has many
pharmacological properties. We showed that BGP suppressed the expression of histamine H1 receptor and IL-9
genes. Here, we sought to investigate the effect of BGP on the suppression of IL-33 gene expression andidentify the responsible active compound. The BGP ethanolic extract suppressed PMA-induced IL-33 gene upregulation in Swiss3T3 cells. It was fractionated using various solvents. The active fraction, then, was further
fractionated by silica gel column chromatography, followed by Sephadex LH-20 column. A single compound was
detected in TLC, and FTIR and NMR analyses identified the compound as betuletol. Betuletol
suppressed PMA-induced IL-33 gene up-regulation. Immunoblot analysis showed that betuletol suppressed
PMA-induced ERK phosphorylation. In conclusion, betuletol was identified as an active compound, and it
suppresses IL-33 gene expression through the inhibition of ERK phosphorylation.

Copyright © The Japanese Pharmacological Society3-B-P-245

一般演題（ポスター）

Restoration of liver sinusoidal endothelial cells after monocrotaline-induced liver
injury〇伊藤 義也1、大高 史聖3、田邉 美奈2、長田 真由子2、山下 敦2、古江 明子2、細野 加奈子1,2、畑中 公1、天野 英樹1,2
北里大・医、2北里大・院医療・分子薬理学、3北里大・医・消化器内科学

1

Hepatic sinusoidal obstruction syndrome (SOS) induced by chemotherapy or hematopoietic stem celltransplantation causes severe liver injury. Although the pathology of SOS is characterized by damage to liver
sinusoidal endothelial cells (LSECs), the processes underlying LSEC repair are incompletely understood. The
purpose of this study was to clarify the regeneration process of damaged hepatic sinusoidal endothelial cells in
SOS.
A SOS model was created by intraperitoneal administration of monocrotaline (MCT) to
male C57BL/6 mice. ALT levels peaked at 48 h post-MCT administration and necrosis occurred around thecentral veins, but recovered to normal at 120 h. The hepatic expression of EC damage markers increased after
48 h and was also accompanied by intrahepatic hemorrhage. Thereafter, EC damage markers decreased with
hepatic repair, and the hepatic sinusoidal structure was reconstructed. LYVE-1 mRNA expression, an LSEC
marker, increased during the liver repair phase. LYVE-1 expression around the central veins was weak before
treatment and further decreased at 48 h, but increased at 96 h and 120 h. Bone marrow-derived cells did notincorporate into the restored LYVE-1-positive cells.
These results suggested that the regenerating endothelium was thought to be due to endothelial
proliferation in the vicinity of the injury, and that bone marrow-derived cells are unlikely to be involved.

Copyright © The Japanese Pharmacological Society3-B-P-246

一般演題（ポスター）

Administration of a SIRT1 activator preserves autophagic activity and improves
age-related sarcopenia in mice.

〇細田 隆介、久野 篤史、中島 龍汰、岩原 直敏、野島 伊世里、堀尾 嘉幸
札幌医科大・医[Background] Sarcopenia is characterized by loss of skeletal muscle function and mass associated with aging.
Autophagy is positively regulated by an NAD+-dependent protein deacetylase SIRT1 and has been reported tomaintain skeletal muscle. In this study, we examined the effect of an activator of SIRT1, resveratrol (RSV), on
muscle autophagy and age-related sarcopenia.
[Method and Results] Ddy mice were fed a normal diet or a diet containing RSV (0.4 g/kg diet) for 37 weeks
starting at 23 weeks of age. Although rotarod running time gradually shortened with aging in both groups, it was
longer in the RSV-fed group at 50 weeks of age. At 60 weeks of age, we sampled tibialis anterior muscles forfurther analyses in control mice (60 wo) and RSV-treated mice (60 wo+RSV). Compared with mice at 20 weeks
of age (20 wo), myofiber diameter determined by HE staining was reduced in 60 wo; however, it was maintainedin 60 wo+RSV. Western blot analysis using anti-acetyl-lysine antibody showed increases in acetylated proteinsin 60 wo compared to 20 wo, suggesting suppression of SIRT1 activity in 60 wo. However, RSV blocked theaging-induced protein acetylation. LC-II/LC3-I ratio, a marker of autophagic activity, was lower and protein
level of p62, which degraded by autophagy, was higher in 60 wo than those in 20 wo, suggesting agingassociated suppression of autophagy. These aging-associated changes were attenuated by RSV treatment.
[Conclusion] These results suggest that activation of SIRT1 in skeletal muscle preserves autophagic activity andattenuates age-related sarcopenia.

Copyright © The Japanese Pharmacological Society3-B-P-247

一般演題（ポスター）

The role of calcitonin gene-related peptide (CGRP) in regulation of intestinal fat
absorption〇細野 加奈子1,2、伊藤 義也1,2、別當 朋広3、畑中 公1,2、馬嶋 正隆1,4、天野 英樹1,2
1

北里大・医、2北里大・院医療・分子薬理、3北里大・医・消化器内科、4神奈川工科大・健康医療科学

Lacteals, lymphatic vessels located at the center of each intestinal villus, play an important role in fat absorption.
Their abnormalities in the structure and function may cause impaired lipid circulation, leading to obesity andabnormal lipid metabolism. We examined whether CGRP (calcitonin gene-related peptide) and its receptor,
RAMP1 (receptor activity modifying protein 1), are involved in fat absorption via lacteals.
RAMP1 knockout (RAMP1 KO) and wild-type (WT) mice were fed a high-fat diet (HFD) or a normal diet(ND) for 8 weeks from 4 to 12 weeks of age. RAMP1KO mice given HFD increased body weight, adipose tissue
mass, serum levels of total cholesterol, triglycerides, and blood glucose, and developed severe obesity compared
to WT mice. In addition, the length of the lacteals tended to be shorter in HFD-fed RAMP1KO mice than in
HFD-fed WT mice. On the other hand, no difference was observed when eating ND. HFD-fed RAMP1KO mice
had decreased expression of lymphatic endothelial markers and pro-lymphangiogenic factors. After oral
administration of BODIPY-FL-labelled C16 fatty acids, the fluorescence intensity in blood in HFD-fed
RAMP1KO mice was higher than that in HFD-fed WT mice.
These results suggest that CGRP/RAMP1 signaling is involved in the regulation of dietary fat absorption by thelacteals.

Copyright © The Japanese Pharmacological Society3-B-P-248

一般演題（ポスター）

In vivo imaging techniques to evaluate fatty liver in a diet-induced NAFLD modelusing PXB mice

〇堀本 泰弘1,2、西方 龍太郎1,2、笹木 祐司3、林田 健一郎1、沼田 洋輔1、角崎 英志4
新日本科学・薬効薬理研究部、2新日本科学・実験動物管理部、3新日本科学・病理研究部、4新日本科学・前臨床カン

1

パニーNonalcoholic fatty liver disease (NAFLD) is now recognized as the most common liver disease in the world.
Nonalcoholic steatohepatitis (NASH), the most advanced form of NAFLD, can progress to cirrhosis andhepatocellular carcinoma. Computed tomography (CT) and magnetic resonance imaging (MRI) are being used
as non-invasive diagnosis for the fatty liver in NAFLD patients. The present study utilized CT and MRI to
examine liver fat mass in human liver chimeric mice (PXB mice, PhoenixBio, Co, Ltd.) fed a choline-deficient,
L-amino-acid-defined, high-fat-diet (CDAHFD, A06071302, Research Diets, Inc.) for 28 and 85 days in orderto produce a diet-induced NAFLD. Serum chemistry and liver histopathology were also conducted. CT and MRI
revealed that the CDAHFD-fed PXB mice (model mice) had higher levels of fat content in the liver compared tothe control diet-fed PXB mice from 14 to 56 day. In serum, the model mice had higher levels of total bilirubincompared to the control mice. Histopathological examinations revealed that Picro-Sirius Red-positive collagenfibers were observed at 28 and 85 days in the control mice and model mice. These results indicate that
CDAHFD can induce fatty liver in PXB mice as seen in human NAFLD and that CT and MRI may be useful
non-invasive methods for evaluation of liver fat in mice.

Copyright © The Japanese Pharmacological Society3-B-P-249

一般演題（ポスター）

Effects of lactoferrin on a choline-deficient methionine-defined High-Fat Diet
(CDAHFD60)-induced NASH model mice

〇古川 恵1、青木 亮憲1、石戸 健太郎2、日塔 武彰2、大野 恵3、出雲 信夫2,4
横浜薬科大・薬・薬学教育セ、2横浜薬科大・薬・薬物治療学研、3(株)NRLファーマ、4横浜薬科大・薬・総合健康メディカ

1

ル研究セ

Inflammation is critical step for nonalcoholic steatohepatitis (NASH) development that progress cirrhosis andliver cancer. Lactoferrin (LF) is known a multifunctional protein such as anti-allergy, anti-inflammatory andanti-obesity effects. However, there is unclear understanding of how LF affects NASH. Therefore, in this study,
we investigated the effect of LF on inflammation in NASH model mice.
NASH model mice were generated by fed a choline-deficient methionine-defined high-fat diet (CDAHFD60) in
male C57BL/6J mice. Mice were randomly assigned to 4 groups: standard diet; CDAHFD60; CDAHFD60 plus
2400 mg/kg LF; CDAHFD60 plus 4800 mg/kg LF. LF was mixed with drinking water and allowed to be freely
taken for 15 days. At the end of experiment, the serum samples and liver tissues were collected. AST and ALT inserum were measured by assay kit and inflammatory cytokines (TNF-α, IL1β and IL-6) in liver were examinedby RT-qPCR.
The increase in AST and ALT induced CADHFD60 was significantly reduced by LF. Furthermore TNF-α, IL-1β and IL-6 expression levels in liver were increased by CADHD60, LF significantly suppressed that. Our
results indicated that LF inhibited NASH development by suppressing inflammatory cytokines.

Copyright © The Japanese Pharmacological Society3-B-P-250

一般演題（ポスター）

Immunohistochemical characterization of TRPV2 and TRPV1 in trinitrobenzene
sulfonic acid (TNBS)-induced colitis in ratsl with visceral hypersensitivity

加藤 伸一、松本 健次郎、安田 浩之、〇宮本 早也佳
京都薬科大・薬・薬物治療学

TRPV2 and TRPV1 are originally identified as heat sensitive TRP channels. We investigated the expression
patterns of TRPV2 and TRPV1 in the rat distal colon and extrinsic primary afferent neurons, and their roles invisceral hypersensitivity in TNBS-induced colitis rats. Both TRPV2 and TRPV1 expressions in colon, DRG, andnodose ganglion (NG) were significantly upregulated in TNBS-induced colitis. TRPV2-expressing cell bodies
colocalized with the intrinsic primary afferent marker NeuN and the inhibitory motor neuronal marker nNOS inmyenteric plexus. TRPV2 expressions also colocalized with the resident macrophage marker ED2 in the mucosa
while no TRPV1-expressing cell bodies were detected in the myenteric plexus. Both TRPV2- and TRPV1expressing cell bodies in DRG and NG were double-labeled with the neuronal retrograde tracer fluorescentfluorogold. Large- and medium-sized TRPV2-positive neurons were labeled with the A-fiber marker NF200,
CGRP, and substance P in DRG while small-sized TRPV1-positive neurons were labeled with the C-fiber
markers IB4, CGRP, and substance P. TRPV2- and TRPV1-positive NG neurons were
2/2
labeled with NF200 and IB4. TNBS treatment increased p-ERK1/2-positive cells in TRPV2 and TRPV1
neurons but did not affect the subpopulations in the DRG and NG. Both TRPV2 and TRPV1 antagonists
significantly alleviated visceral hypersensitivity in TNBS-induced colitis model rats. These findings suggest that
intrinsic/extrinsic TRPV2- and extrinsic TRPV1-neurons contribute to visceral hypersensitivity in anexperimental colitis.

Copyright © The Japanese Pharmacological Society3-B-P-251

一般演題（ポスター）

Analysis of drug-induced appendicitis using the Japanese Adverse Drug Event
Report (JADER) database〇京谷 陽司、趙 晶、中平 毅一、吉栖 正典
奈良県立医科大・医[Introduction] Appendicitis is one of the most frequent diseases affecting people of all ages. Several drugs werereported as suspects for appendicitis in Japanese Adverse Drug Event Report (JADER) database, but
appendicitis is not listed in the package inserts of these drugs except barium sulfate. We investigate theassociation of these drugs with appendicitis.
[Methods] We assessed the JADER database between Jan. 2004 and Dec. 2021. Signal was detected when thelower limit of the 95% CI of the reporting odds ratio (ROR) was greater than 1. Time-to-onset profile and ROR
adjusted for age and sex were detected by weibull plot and logistic regression analysis, respectively.
[Results] The total number of adverse event reports was 1,551,265, of which 591 were reports of appendicitis.
Signals were detected in 22 drugs except barium sulfate. Among them, 8 drugs showed significant shapeparameters (ß) in weibull plot and drug-dependent patterns for changes in the risk of appendicitis over time.
Adjusted RORs indicated significant association for 5 drugs.
[Conclusion] Our results suggest that several drugs are associated with appendicitis. Among them, thepharmacological mechanism of clozapine is still unclear, and further investigation of the mechanism of clozapine
might contribute to establish the management and control of appendicitis.

Copyright © The Japanese Pharmacological Society3-B-P-252

一般演題（ポスター）

L-DOPA suppresses bladder smooth muscle contraction induced by carbachol in
rats〇ZOU SUO1、清水 孝洋1、濱田 知里1、清水 翔吾1、東 洋一郎1、増川 太輝2、五嶋 良郎2、齊藤 源顕1
1

高知大・医・薬理、2横浜市立大・院医・薬理L-3,4-Dihydroxyphenylalanine (L-DOPA) has been recognized as a precursor of dopamine. Recently, L-DOPA
has been proposed as a neurotransmitter and is reported to sensitize vasomotor response to sympathetic tone
through activation of L-DOPA receptor GPR143. In this study, to clarify roles of L-DOPA/GPR143 signaling in
regulation of parasympathetic tone, we examined effects of L-DOPA on muscarinic receptor-mediated bladdercontraction in wild-type (WT) andgene-deficient (-/y) rats. Bladder strips were prepared fromnine-week-old male rats, and effects of pretreatment with L-DOPA (10-8, 10-7 and 10-6 M) on carbachol (CCh,10-8 to 3×10-4 M)-induced bladder strips contraction were investigated by organ bath study. In bladder strips
from WT rats, L-DOPA alone showed no contraction or relaxation, while pretreated L-DOPA dose-dependently
suppressed CCh-induced contraction. The L-DOPA-induced suppression was also observed in bladder strips
from

-/y

rats. These results suggest that L-DOPA induces suppression of the muscarinic receptor-

mediated rat bladder contraction independently of GPR143 signal transduction.

Copyright © The Japanese Pharmacological Society3-B-P-253

一般演題（ポスター）

Inhibition of CD38 promotes muscle regeneration〇夜久 圭介1、Nawaz Allah1、中川 崇1,2
富山大・医・分子医科薬理、2富山大・未病研究センター

1

Sarcopenia is aging-related progressive loss of skeletal muscle mass and strength, which is associated withdecreased quality of life and increased mortality. Aging-related decline of muscle regeneration is a risk factor forthe onset of sarcopenia.
Nicotinamide adenine dinucleotide (NAD+) is considered as an anti-aging molecule which can suppress theonset of aging-related diseases. NAD+ is an essential molecule for all living organisms, which is associated withmultiple biological processes, including energy production, DNA repair, and protein modifications. NAD+ levels
in tissues decrease with aging, which leads functional loss of tissues. On the other hand, NAD+ repletion isconsidered to have therapeutic potential against aging-related diseases. For example, treatment of NAD+precursor is reported to suppress loss of muscle function caused by aging, which is lead by the increase in NAD+levels.
Increased degradation of NAD+ is the cause of decreased NAD+ levels during aging. CD38 is the enzyme which
degrades NAD+, and expression levels of CD38 increase with aging. NAD+ decline caused by CD38 leads
multiple aging signatures, such as delayed metabolism, cell senescence, and tissue fibrosis. Importantly,
inhibition or genetic loss of CD38 can remarkably increase NAD+ levels in tissues. Additionally, inhibition of
CD38 improves muscle function in aged animals. Therefore, CD38 is an attractive target against the onset ofsarcopenia.
In this research, we investigated the effect of inhibition or genetic loss of CD38 on muscle regeneration. Genetic
deletion of CD38 reduced muscle fibrosis and improved recovery of muscle after the treatment of cardiotoxin.
Furthermore, inhibition of CD38 increased NAD+ levels and promoted myoblast differentiation. These results
suggest that inhibition of CD38 has therapeutic potential against the onset of sarcopenia.

Copyright © The Japanese Pharmacological Society3-B-P-254

一般演題（ポスター）

Angiotensin II-induced contraction of superior mesenteric arteries fromstreptozotocin-induced diabetic rats

〇寺田 侑加1、日比野 凪沙1、井上 滉太1、藤田 義人2、稲垣 暢也2、屋山 勝俊1
1

神戸学院大・薬・循環器薬理、2京都大・院医・糖尿病・内分泌・栄養内科学

Diabetes is a major risk factor for vascular diseases, and often associated with vascular dysfunction. However,
the mechanisms of vascular dysfunction in diabetes are not fully understood. Inactivation of myosin light chain
phosphatase thorough myosin phosphatase targeting subunit 1 phosphorylation by Rho kinase is important forangiotensin II (Ang II)-induced contraction. We have previously reported that Rho kinase activation by Src,
epidermal growth factor receptor (EGFR), extracellular signal-regulated kinase 1 and 2 (Erk1/2) is involved inprotein tyrosine phosphatase inhibitor-induced contraction. We therefore investigated the effects of inhibitorsof these factors on Ang II-induced contraction in diabetic rat superior mesenteric arteries.
Wistar male rats were induced diabetes by intraperitoneal injection of streptozotocin (40 mg/kg). After 8 weeks,superior mesenteric arteries were excised and performed organ chamber experiments. Protein levels were
measured by Western blotting.
The contraction of superior mesenteric arteries from diabetic rats increased in the response to Ang II incomparison with those from the control rats. However, we did not observe any differences in Ang II-induced
contraction between the groups in the presence of Rho kinase, EGFR, and Erk1/2 inhibitors. Moreover, protein
level of Erk1/2 was significantly increased in diabetic rats. These data suggest that increased activity of Rho
kinase, EGFR, and Erk1/2 might be involved in excessive contraction by Ang II in superior mesenteric arteriesof diabetic rats.

Copyright © The Japanese Pharmacological Society3-B-P-255

一般演題（ポスター）

Endothelium-dependent and -independent Vasodilator Effects of SGLT2
inhibitors〇金田 剛治1、大山 唯花1、佐々木 典康2、金田 寿子1,3
日本獣医生命科学大・獣医・獣医薬理研究室、2日本獣医生命科学大・獣医・獣医生化学研究室、3帝京大・医・解剖学

1

講座

SGLT2 inhibitors (SGLT2i) are new agents for patients with heart failure (HF) irrespective of diabetes.
However, the effects of in vasoconstriction remain unclear. This study examined the mechanism ofvasorelaxation induced by SGLT2i in endothelium-intact and -denuded rat aorta and mesenteric artery. 1)
Dapagliflozin and empagliflozin inhibited phenylephrine (PE)-induced contraction in a dose-dependent
manner. However, this relaxation was lower in the absence of the endothelium in aorta and mesenteric artery. 2)
Increase in dapagliflozin and empagliflozin-induced relaxation in the presence of the endothelium wasattenuated by preincubation in L-NAME (100 μM) and the removal of the endothelium, but not indomethacin(10 μM) in aorta and mesenteric artery. 3)Remove of glucose from PSS did not affect dapagliflozin andempagliflozin -induced relaxation in endothelium-denuded aorta. 4) In fura 2-loaded endothelium-denuded
aorta, empagliflozin inhibited high K+- or PE-induced muscle tension and increase of intracellular Ca2+ ([Ca2+]i)level. These results may suggest that dapagliflozin and empagliflozin causes vasorelaxation by increasing cGMP
content in correlation with the release of NO from endothelial cells and by decreasing [Ca2+]i) level via
inhibiting voltage-dependent Ca2+ channel.

Copyright © The Japanese Pharmacological Society3-B-P-256

一般演題（ポスター）

ATP-evoked relaxation of esophageal smooth muscle via potassium channels inrats〇椎名 貴彦、志水 泰武
岐阜大・応用生物・獣医生理

The external muscle layer of the mammalian esophagus consists of striated muscle fibers and smooth muscle
fibers. Striated muscle is mainly regulated by cholinergic signaling, whereas smooth muscle is regulated bycholinergic and non-cholinergic signaling in the esophagus. ATP is a representative non-cholinergic
extracellular transmitter, which control smooth muscle motility in the blood vessels and gastrointestinal tracts
via purinergic receptors. We have demonstrated that exogenous application of ATP evokes relaxation of smoothmuscle in the muscaris mucosa of the rat esophagus. In the present study, the aim was to clarify
involvement of potassium channels in purinergic relaxation of the esophageal smooth muscle. An isolated
segment of the rat esophagus was placed in an organ bath and the mechanical responses were recorded using aforce transducer. After contraction of esophageal smooth muscle was induced by carbachol, we applied ATP,which evoked relaxation of smooth muscle. On the other hand, ATP did not affect high-potassium induced
contraction. Pre-application of an antagonist of ATP-dependent potassium channels (KATP channels) blocked
ATP-induced relaxation, but not voltage-gated potassium channel blockers. These findings indicatethat KATP channels might be involved in ATP-induced relaxation of the esophageal smooth muscle in rats.

Copyright © The Japanese Pharmacological Society3-B-P-257

一般演題（ポスター）

Effect of cisplatin on the expressions of myosin heavy chain isoforms in skeletal
muscle of mouse〇酒井 寛泰1、Xu Xinran1、宮内 優1、千葉 義彦2、今 理紗子1、五十嵐 信智1、亀井 淳三1,3、細江 智夫1,4
1

星薬科大・薬、2星薬科大・薬・分子生物学、3順天堂大・企画調査室、4星薬科大・薬・生物制御科学

We previously reported that cisplatin induced skeletal muscle atrophy in mice. Myosin heavy chain (MyHC) is
one of the structural proteins essential for the contractile response of skeletal muscle, and several isoforms exist.
However, the kinetics of its expression in cisplatin-induced muscle atrophy is unknown. In this study, weinvestigated the expression changes of MyHC isoforms in cisplatin-induced muscle atrophy. Mice were treatedwith cisplatin intraperitoneally once daily for 4 days, and quadriceps muscles were isolated 1 day after the last
dose; C2C12 myotubes were also treated with cisplatin. Cisplatin significantly reduced protein expression of
MyHC isoforms (I, IIa, IIx, IIb) in mice compared to controls. However, only MyHC-IIa was downregulated
when changes in MyHC gene expression were examined. Similar to the results in mice, cisplatin attenuatedprotein expression of all MyHC isoforms in C2C12 myotubes, and gene expression of MyHC-IIa wasdownregulated. Furthermore, the downregulation of protein expression of all MyHC isoforms was inhibited bytreatment with MG-132, a proteasome inhibitor. These results indicate that cisplatin downregulates MyHC
protein expression. Thus, we suggest that proteasomal degradation of MyHC proteins is one of the causes ofcisplatin-induced muscle atrophy.

Copyright © The Japanese Pharmacological Society3-B-P-258

一般演題（ポスター）

Establishment of muscle strength measurement system using aged mice

〇真壁 大地、森田 枝美、緒里 真一、吉原 佐江子、清水 広夢、田代 貴士、片山 誠一、廣中 直行、西 勝英㈱LSIM安全科学研究所・熊本研究所・薬理研究部

Japan has a very high aging rate among developed countries. The increase of medical expenses in a super-agedsociety has become serious. It is important to maintain good quality of life (QOL) throughout life. One of thefactors that lowers QOL of the elderly is a decline in mobility accompanied by a decrease in muscle strength
such as sarcopenia. Many food and drug suppliers are developing products to prevent muscle weakness. In thisstudy, we tried to establish a test system to evaluate effects of therapeutic agents on muscle weakness using aged
mice. First, we compared the muscle weakness of 12, 40, 53, 79, 92, and 105-week-old C57BL/6J male miceusing a small animal muscle strength measuring device (Aurora Scientific). Next, we measured blood and organ
concentrations of AGEs, NT-proBNP, and MURF1, which are indicators of aging. Muscle pathology was also
examined. Compared with 12-week mice, aged mice (79 to 92 weeks) showed significantly lower muscle
strength. Finally, we tested the effects of nicotinamide mononucleotide (NMN, a precursor of NAD), which isrecently attracted attention as a dietary supplement with potential anti-aging effects. Oral administration of
NMN for 12 weeks showed preventive effect on muscle weakness in aged mice. The present test system would
be useful to evaluate potential efficacy of newly developed therapeutic agents on muscle weakness in theelderlies.

Copyright © The Japanese Pharmacological Society3-B-P-259

一般演題（ポスター）
2++

Suppressive effect of the Ca -activated K channel KCa3.1 activator, SKA-121 on
IL-10 and IL-8 expression in THP-1-derived M2 macrophages

松井 未来、梶栗 潤子、鬼頭 宏彰、遠藤 京子、〇大矢 進
名古屋市立大・院医

The human monocytic leukemia cell line, THP-1-differentiated macrophages are a useful tool to investigate thephysiological significance of tumor-associated macrophages (TAMs). In the tumor microenvironment (TME),
TAMs with the M2-like phenotype play a critical role in the promotion of cancer progression and metastasis by
inhibiting the immune surveillance system. We examined the involvement of Ca2+-activated K+ channel KCa3.1inthe expression of pro-tumorigenic cytokines and angiogenic growth factors in THP-1-derived M2 macrophages.
THP-1 cells into M0 macrophages were induced by a treatment with PMA treatment for 24 hr, and then cells
were treated with IL-4 and IL-13 for 72 hr to induce the polarization of M2 macrophages. The expression levelsof IL-8 and IL-10 were significantly decreased by treatment with the selective KCa3.1 activator, SKA-121 in THP-1-derived M2 macrophages. Furthermore, under in vitro experimental conditions that mimic extracellular K+
levels in the TME, IL-8 and IL-10 levels were both significantly elevated, and these increases were reversed bytreatment with SKA-121. Among several signaling pathways potentially involved in the transcriptional
regulation of IL-8 and IL-10, respective treatments with ERK and JNK inhibitors significantly repressed theirtranscriptions, and treatment with SKA-121 significantly reduced the phosphorylated ERK, JNK, c-Jun, andCREB levels. These results strongly suggest that the KCa3.1 activator may suppress IL-10-induced tumor
immune surveillance escape and IL-8-induced tumorigenicity and metastasis by inhibiting their production
from TAMs through both ERK-CREB and JNK-c-Jun cascades.

Copyright © The Japanese Pharmacological Society3-B-P-260

一般演題（ポスター）

Amino acid transporter LAT1 (SLC7A5) on cancer cell-derived exosomes as apotential prognostic- and diagnostic biomarker.

〇大垣 隆一1,2、Liu Yumiao1、徐 旻憓1、岡西 広樹1、金井 好克1,2
大阪大・院医・生体システム薬理、2大阪大・先導的学際研究機構・生命医科学融合フロンティア部門

1

L-type amino acid transporter 1 (LAT1/SLC7A5), which transports large neutral amino acids, is highly
upregulated in various cancer cells and supports their enhanced growth and proliferation. Previous
immunohistochemical studies have reported significant correlations between the high LAT1 expression and thepoor prognosis of patients in multiple cancer types. Furthermore, several LAT1-selective inhibitors are currently
under evaluation as novel anti-cancer drugs. Detecting the LAT1 expression levels in tumor lesions withminimally invasive methods, therefore, would be beneficial for prognosis and diagnosis of cancers, including thecompanion diagnosis in the upcoming LAT1-targeted therapy. Molecules on exosomes, small extracellular
vesicles, released from cancer cells are emerging as a novel class of biomarkers. In this study, we found that
LAT1 is detectable in exosomes isolated from the cell culture supernatants of cancer cells. Notably, theabundance of LAT1 on exosomes was associated with its expression level in the cancer cells from which they
derived. LAT1 was also present in exosomes isolated

from peritoneal washing fluid of intraperitoneal

tumor-bearing mice. These results indicate that LAT1 on cancer cell-derived exosomes holds a significant
potential as a biomarker for diagnosis and prognosis of cancers.

Copyright © The Japanese Pharmacological Society3-B-P-261

一般演題（ポスター）

The effect of acute treatment with cyclophosphamide in post-weaning on stressresponse and hippocampal stem/progenitor cell proliferation in the adulthood〇尾中 勇祐、山口 太郎、米山 雅紀
摂南大・薬Chemotherapy for childhood cancer can cause late-appearing side effects in survivors that affect multiple
organs, including the brain. Cyclophosphamide is used in childhood cancer and has the blood brain barrier
permeability. The present study aims to reveal whether an acute treatment with cyclophosphamide after
weaning affects the brain function and hippocampal neurogenesis in the adult mouse. Cyclophosphamide wereintraperitoneally once injected to 3 weeks old male ddY mice. Five weeks after injection, restraint stress loadingor the behavioral test battery was performed. After the test battery, mice were received 2 consecutive injectionsof 5-bromo-2′-deoxyuridine (BrdU) with a 12-h interval. BrdU positive cells in the hippocampal dentate gyrus
were counted after immunostaining for BrdU. Treatment with cyclophosphamide had the ability to enhance
restraint stress-induced increase of plasma corticosterone, and decrease the number of BrdU-positive cells inthe dentate gyrus 5 weeks afterward. However, cyclophosphamide failed to change emotional and cognitive
functions. These data suggest that the acute treatment with cyclophosphamide in post-weaning exhibits
prolonged changes in stress response and the suppressive effect on hippocampal neurogenesis.

Copyright © The Japanese Pharmacological Society3-B-P-262

一般演題（ポスター）

L-type amino acid transporter 1 inhibitor JPH203 as a new therapeutic target for
castration resistant prostate cancer treatment〇齋藤 心平1,2、坂本 信一2、濱口 紀江1、齊藤 将太1、裵 祥存1,2、霊園 良恵1、平山 友里1、橋本 弘史1、市川 智彦2、安西 尚彦1
千葉大・院医薬・薬理学教室、2千葉大学医学部附属病院・泌尿器科

1

L-type amino acid transporter 1 (LAT1) plays a role in transporting essential amino acids including leucine,which regulates the mTOR signaling pathway. Here, we studied the expression profile and functional role of
LAT1 in prostate cancer using JPH203, a specific inhibitor of LAT1. LAT1 was highly expressed in castrationresistant prostate cancer (CRPC) cell lines including C4-2 and PC-3, while poorly expressed in castration
sensitive LNCaP cells. 5 μ M of JPH203 significantly blocked 14C leucine uptake in CRPC cells, while notaffected in LNCaP cells. JPH203 inhibited cell migration at 30 μ M in CRPC cells, while not affected in LNCaP
cells. Combination of JPH203 30 μ M and Enzalutamide 10 μ M additively blocked the cell proliferation in
CRPC cells. RNA sequence identified CD24 as a novel downstream target of JPH203 in C4-2 cells. SiCD24
blocked cell migration in C4-2 cells via blocking phosphorylation of GSK3β and activating phosphorylation of
β catenin. JPH203 25mg/kg significantly inhibited tumor growth in the C4-2 xenograft nude mouse model.
Targeting LAT1 by JPH203 may represent a novel therapeutic option in CRPC.

Copyright © The Japanese Pharmacological Society3-B-P-263

一般演題（ポスター）

Eribulin modulates stathmin dynamics and enhances paclitaxel sensitivity in
ovarian cancer cells〇安曇 麻奈、吉江 幹浩、高野 航瑠、草間 和哉、田村 和広
東京薬科大・薬・内分泌薬理

Ovarian cancer is the major cause of death from gynecologic malignancies owing to poor prognosis. Stathmin, amicrotubule-destabilizing protein, is expressed in a stage-dependent manner in ovarian cancer. Eribulin is a
microtubule dynamics inhibitor used to treat breast cancer and sarcoma. The present study explored theantitumor efficacy of eribulin and the involvement of stathmin in the action of eribulin in ovarian cancers. In a
xenograft model of ovarian cancer, eribulin treatment reduced the tumor weight accompanied with an increased
level of phosphorylated stathmin, and a decreased expression of proliferating cell nuclear antigen (PCNA).
Eribulin abolished the formation of tumor blood vessels with large lumens. In cultured ovarian cancer cell lines,
eribulin stimulated stathmin phosphorylation and decreased stathmin protein expression. A protein phosphatase
2A (PP2A) activator FTY720 attenuated the eribulin-induced phosphorylation of stathmin. The levels of PP2A
subunits were downregulated by eribulin. SiRNA-mediated stathmin knockdown attenuated the inhibitory
effects of eribulin on cell viability. In addition, eribulin enhanced the antiproliferative effects of paclitaxel on
ovarian cancer cells, which was accompanied by a decreased stathmin expression. These results suggest that
eribulin may suppress ovarian cancer tumorigenesis partly by regulating the stathmin dynamics, and thatcombined treatment of eribulin with paclitaxel could be useful for ovarian cancer.

Copyright © The Japanese Pharmacological Society3-B-P-264

一般演題（ポスター）

Differentiation-inducing factor-1 inhibited cancer cell adhesion to vascularendothelial cells via curtailing VCAM-1 protein synthesis〇有岡 将基、石兼 真、笹栗 俊之、高橋 富美
産業医科大・医・薬理学

We previously reported that differentiation-inducing factor-1 (DIF-1) inhibited lung colony formation in mousemodels of cancer metastasis by suppressing cancer cell proliferation and motility. Although adhesion of
circulating tumor cells to vascular endothelial cells is an essential process for the initiation of tumor metastasis
formation, the effects of DIF-1 on this process has not been elucidated. In this study, therefore, we investigatedthe effect of DIF-1 on the adhesion ability of tumor cells (human melanoma cells A2058, mouse melanoma cells
B16BL6 and human colon cancer cells HCT116) to human umbilical vein endothelial cells (HUVECs).
Treatment with DIF-1 significantly inhibited these cancer cells adhesion to HUVECs. To elucidate themechanism of DIF-1 action, we analyzed adhesion-related genes in HUVECs. We found that the expression ofvascular endothelial cell adhesion molecule-1 (VCAM-1) was suppressed by DIF-1 and the protein synthesis
inhibitor cycloheximide abolished this inhibitory effect. We further evaluated the effect of DIF-1 using the lungmetastasis model. Surprisingly, only 3 days administration of DIF-1 prior to inoculation of B16BL6 melanoma
cells significantly inhibited lung colony formation. These results suggest that the main mechanism of DIF-1
anti-metastatic effect might be attenuation of adhesion between circulating tumor cells to blood vessels.

Copyright © The Japanese Pharmacological Society3-B-P-265

一般演題（ポスター）

The BDNF/TRKB pathway promotes EMT to induce parotid gland cancer cellaggressiveness via interaction with CAF〇森脇 一将1、桑原 宏子2、綾仁 悠介3、東野 正明3、寺田 哲也3、河田 了3、朝日 通雄11

大阪医科薬科大・医・薬理、2大阪医科薬科大・医・病理、3大阪医科薬科大・医・耳鼻咽喉科

The molecular features of parotid gland cancer (PGC) are not fully understood enough to develop an effective
drug therapy because of the rarity. Given the poor prognosis of many human cancers in which TRKB is highly
expressed, we investigated the involvement of the BDNF/TRKB pathway in PGC tissue using clinicalspecimens and observed high expressions of TRKB and BDNF in both tumor cells and stromal cells such as
cancer-associated fibroblasts (CAFs). Therefore, to obtain more detail information of BDNF/TRKB signaling in
PGC, we established primary co-culture system of patient-derived PGC cells and CAFs. In the culture system,
PGC cells co-cultured with CAFs exhibited significant upregulation of BDNF and epithelial-mesenchymaltransition (EMT). Similar results were observed in PGC cells treated with conditioned medium (CM) from coculture of PGC cells and CAFs. TRK inhibitors suppressed BDNF- or CM-induced Snail upregulation and cell
migration in PGC cells. Importantly, immunohistochemical and clinicopathological analyses of tumors from thepatients with PGC revealed that TRKB expression levels in PGC cells were significantly correlated withaggressive features, including vascular invasion, nodal metastasis, and poor prognosis. Collectively, these datasuggest that the BDNF/TRKB pathway regulates PGC cell aggressiveness via cross-talk with CAFs and is apotential therapeutic target for PGC harboring invasive and metastatic features.

Copyright © The Japanese Pharmacological Society4-B-P-266

一般演題（ポスター）

Correlational changes in insular cortical neuronal activity and cardiac signals〇木下 航輔1、久我 奈穂子2、佐々木 拓哉2,3
1

東北大・薬・薬理、2東北大・院薬・薬理、3東京大・院薬・薬品作用The insular cortex serves as a hub cortical region that is bidirectionally connecting to an extensive cortical andsubcortical brain areas and plays a crucial role in interoception, the sensation of internal states of the body, suchas heartbeats, hunger, and blood pressure. However, neurophysiological mechanisms and insights supporting
this hypothesis remain to be clarified. To address this issue, we performed simultaneous recordings of multiunit
spike patterns and local field potential (LFP) signals from the insular cortex, an electrocardiogram signal, and aperipheral blood glucose concentration from freely moving rats. Recordings were daily obtained for seven hours.
No pronounced temporal correlational changes were found between ongoing heart rate and blood glucose
fluctuations. Insular cortical LFP power in some frequency bands, such as the delta and theta bands, showedapparent temporal correlational changes with ongoing heart rates when the rats were resting. At single-celllevels, a subset of insular cortical neurons increased or decreased their spike rates in response to changes inheart rates and blood glucose levels. These results highlight insular cortical neurons as a detector of temporalchanges in interoceptive signals.

Copyright © The Japanese Pharmacological Society4-B-P-267

一般演題（ポスター）

Alteration in BAK (BCL2 antagonist)1 and Bcl-2-associated athanogene (BAG) 3
in Hypothyroid Embryonic Chick Cerebellum〇三部 篤、猪俣 結衣、髙橋 慎太郎、玉田 さち、夏堀 陽子、東尾 里英子
岩手医科大・薬Hypothyroid conditions during fetal development can induce severe developmental defects such as mental
retardation. Thyroid dysfunction and hypothyroid conditions induced by anti-thyroid drugs may influence
neuronal development during embryogenesis. Little is known, however, about the details of the mechanisms
underlying the adverse effects of thyroid dysfunction on neuronal development. In this study, we examined thegene expression levels of Bcl-2 family members and Bcl-2-associated athanogene (BAG) 3 in chick cerebellum
under hypothyroid conditions using a fertilized chicken egg/chick embryo model. The hypothyroid chicks
exhibited a marked reduction in BAG3 protein levels concomitant with a marked increase in mitochondrial Bak1
protein levels in cerebellum while no alteration in gene expression of Bak1 was seen. Since evidence of apoptotic
cell death was observed in the cerebellum under hypothyroid conditions, these results suggest that reduction in
BAG3 as well as enhancement of Bak1 protein may play an important role in the apoptosis of neuronal cells
under hypothyroid conditions .

Copyright © The Japanese Pharmacological Society4-B-P-268

一般演題（ポスター）

Antioxidative activity of remifentanil as a direct free radical scavenger〇徳丸 治1、山崎 玲音2、尾方 和枝1、松本 重清3、北野 敬明3
大分大・福祉健康科学・生理、2大分大・医・学生、3大分大・医・麻酔1

Remifentanil is an ultra-short acting opioid used for general anesthesia. Recently, it is reported that remifentanil
is preventive against oxidative stress including ischemia/reperfusion injury and inflammation. However, detailed
mechanism is not fully described. We aimed to study free radical scavenging activity of remifentanil against
multiple free radicals.
Free radical scavenging activity of remifentanil was evaluated against nine species of free radicals by electron
spin resonance spectroscopy with the spin-trapping method. From dose-response curves, reaction rate constants
with free radicals examined were estimated. Antioxidative activity against lipids was assessed by TBARS assay.
Remifentanil significantly scavenged the following five free radical species in dose-dependent manners; hydroxyl
radical,-butoxyl radical, ascorbyl free radical, singlet oxygen and nitric oxide. Remifentanil did not scavenge

superoxide anion,-butyl peroxyl radical, DPPH and tyrosyl radical. Remifentanil significantly inhibited

oxidation of lipids in mice brain tissue.
Remifentanil dose-dependently scavenged multiple free radicals including hydroxyl radical. It is speculated thatthe direct free radical scavenging activity of remifentanil might contribute to its antioxidative activity
and.

Copyright © The Japanese Pharmacological Society4-B-P-269

一般演題（ポスター）

Involvement of Mas-related G protein-coupled receptor X1 (MrgprX1) in thechronic itch

泉本 直樹、〇今野 光洋、森山 正樹、湯沢 夏美、岩村 智勝、成見 英樹
東レ・医薬研究所・創薬薬理研究室

Atopic dermatitis (AD) is a common dermatologic disease that is accompanied by severe chronic pruritus.
Recently, it has been reported that mas-related G protein-coupled receptor (Mrgpr) involves in the control ofhistamine-independent itch and the modulation of Mrgpr might be a promising target for the treatment ofchronic itch. In the present study, we used the transgenic mice expressed human MrgprX1 in the sensory nerveand Mrgpr cluster KO (Mrgpr-KO) mice and evaluated the scratching behaviors in the acute itch model evokedby BAM8-22. In addition, the modulation of Mrgpr and the effect of steroid drug in the MC903-induced chronic
itch model in both transgenic mice were examined. Intradermal BAM8-22 did not induce the scratching
behaviors in the Mrgpr-KO mice over vehicle-treatment, on the other hand, it evoked scratching behaviors in
the hMrgprX1 transgenic mice compared to the vehicle-treated animals. In the chronic itch model evoked by therepetitive application of MC903 produced the scratching behaviors in both Mrgpr-KO and hMrgprX1
transgenic mice over vehicle-treatment, but the ones in the hMrgprX1 mice evidently outweighed them in the
Mrgpr-KO mice. In addition, dexamethasone, a steroid drug, significantly attenuated the scratching behaviors
evoked by MC903 in both Mrgpr-KO and hMrgprX1 transgenic mice to the same extent. From theseobservations, it is suggested that the itch in the AD patients might be able to be inhibited completely by the
combination of steroid and hMrgprX1 antagonist.

Copyright © The Japanese Pharmacological Society4-B-P-270

一般演題（ポスター）

Mechanisms for the allodynia caused by complement anaphylatoxin C5a in mice

〇田島 和樹1
近畿大・薬・病態薬理学研究室、2岡山大・院医歯薬・薬理、3岡山大・院医歯薬・創薬研究推進、4旭化成ファーマ・医薬

1研究センター・薬理The thrombin-thrombomodulin (TM) complex generates activated forms of protein C (APC) and thrombinactivatable fibrinolysis inhibitor (TAFIa), known as carboxypeptidase B (CPB), and degrades high mobility
group box 1 (HMGB1), a pronociceptive molecule. We have shown that soluble TM (TMα) prevents
chemotherapy-induced peripheral neuropathy (CIPN) in rodents, and that APC and TAFIa/CPB, as well as
HMGB1 degradation, contribute to the anti-CIPN effect of TMα. Given our recent evidence for the
involvement of complement C5a, degradable by TAFIa/CPB, in CIPN, we analyzed the mechanisms forperipheral C5a-mediated pain in mice. Intraplantar (i.pl.) injection of C5a at 30-300 ng induced dosedependent mechanical allodynia, as assessed by von Frey test, an effect abolished by i.p. administration of
DF2593A, a mouse C5a receptor (C5aR) antagonist, TAFIa/CPB, TMα or an anti-HMGB1-neutralizing
antibody. Systemic (i.p.), but not local (i.pl.), administration of liposomal clodronate, a macrophage depletor,
partially reduced the C5a-induced allodynia. In macrophage-like RAW 264.7 cells, C5a caused HMGB1 releasein the presence, but not absence, of a sub-effective concentration of lipopolysaccharide. Together, C5a-induced
allodynia is considered to involve C5aR-dependent HMGB1 release from monocyte-derived macrophages andnon-macrophage cells.

Copyright © The Japanese Pharmacological Society4-B-P-271

一般演題（ポスター）

In vitro assessment of drug-induced peripheral pain in DRG neurons at single
cell level using CMOS-MEA〇韓 笑波、柴田 未可子、松田 直毅、鈴木 郁郎
東北工業大学・大学院工学研究科

Drug-induced peripheral neuropathy occurs as a major adverse effect of chemotherapy. However, a highly
accurate assessment platform for drug-induced peripheral neuropathy and pain has not been established yet. In
the present study, we introduced a CMOS-MEA system with high spatiotemporal resolution, to detect
electrophysiological activity of cultured DRG neurons.
The CMOS-MEA that consists of 236,880 electrodes, makes it possible to identify each single soma of active
DRG neurons. Therefore, the spike pattern of each single neuron is able to be demonstrated before and after
cumulative additions of TRP channel agonist (i.e., capsaicin, AITC and menthol). And we found that paclitaxel,
a wildly used anticancer drug, could sensitive DRG response to capsaicin burning by spike pattern analysis.
After identifying the axonal conduction pathway, we also calculated the velocity of axonal conduction from eachneuron, and the result showed to be in a reasonable range. The mean conduction velocity could be increased byadministration of vincristine, which is known to cause peripheral neuropathy.
Using CMOS-MEA system with high spatiotemporal resolution, we succeed in evaluating spike pattern at single
neuron level and measuring axonal conduction velocity in cultured DRG neurons. And changes in theseparameters of DRG neuron response to anti-cancer drugs could also be detected, suggesting that this system is
effective for assessment of drug-induced peripheral neuropathy and pain.

Copyright © The Japanese Pharmacological Society4-B-P-272

一般演題（ポスター）

Intracochlear macrophages are involved in the development of sensorineural
hearing loss by decreasing inner hair cell synapses〇山口 太郎摂南大・薬The development of sensorineural hearing loss is a serious problem because it significantly reduces the qualityof life. Age-related, noise-induced, and drug-induced sensorineural hearing loss has been reported to decrease
inner hair cell synapses (IHC synapses) in its early stages. This study aimed to determine the cause of thisdecrease in IHC synapses. We prepared the mice in which repeated exposure (1-h exposure per day, 5 times) tomoderate noise (8-kHz octave band noise, 90 dB sound pressure level) induced gradual hearing impairment
along with a loss of IHC synapses. Treatment with PLX3397 (macrophage-depletion drug, 290 mg/kg)
significantly improved reduced the number of synapses and hearing impairment caused by repeated noise
exposure. Treatment with minocycline (macrophage-activation inhibitor, 50 mg/kg) significantly improved
reduced the number of synapses and hearing impairment caused by repeated noise exposure. These results
suggest that intracochlear macrophages are activated by noise exposure and may negatively regulate the numberof IHC synapses.

Copyright © The Japanese Pharmacological Society4-B-P-273

一般演題（ポスター）

Contractility assessment using hiPSC-CMs with alignment〇佐塚 文乃、林 紗代、柳田 翔太、諫田 泰成
国衛研・薬理部

Evaluation of cardiomyocyte contraction using hiPSC-CMs holds great promise to predict drug-induced heart
failures in vitro. However, it has been difficult to detect the positive inotropic effects of drug using hiPSC-CMs.
Due to the immature property, hiPSC-CMs do not show alignment. In this study, we evaluated whether hiPSCCM with alignment can detect the drug-induced positive inotropic effects. We used hiPSC-CM (iCell
Cardiomyocyte, FCDI), which were cultured using commercially available plates for alignment. After long-term
culture, we found that the aligned culture improved the basal contraction velocity, which were measured bymotion vector analysis (SI8000, Sony). To understand the molecular mechanisms of the contraction
improvement, we performed next-gene sequencing analysis. Gene expression levels of ion channels andmetabolism-related factors were up-regulated by alignment. In addition, we observed that isoproterenol
enhanced contraction velocity in hiPSC-CMs with alignment in a dose-dependent manner. These results
suggests that the aligned culture of hiPSC-CMs facilitates the maturation and the responses to isoproterenol.

Copyright © The Japanese Pharmacological Society4-B-P-274

一般演題（ポスター）

The interaction between MMP-9 and TNF-α contributes to the exacerbation ofcapillary degeneration in a rat model of NMDA-induced retinal injury〇小嶋 美帆香、浅野 大樹、森田 茜、柏原 俊英、中原 努
北里大・薬・分子薬理

Matrix metalloproteinases (MMPs) and tumor necrosis factor (TNF)-α contribute to the pathogenesis ofseveral ocular diseases. In this study, we aimed to determine the role of interaction between TNF-α and MMP-9 in capillary degeneration. In rats, retinal injury was induced by intravitreal injection of

methyl-D-aspartic

acid (NMDA) at postnatal day 7. We examined 1) the effects of blocking MMP-9 and TNF-α signaling pathway
on capillary degeneration, 2) changes in protein levels and distribution of MMP-9 and TNF-α, and 3) theinteraction between MMP-9 and TNF-α in regulating the expression level of each protein in retinas of NMDAinjected eyes. Intravitreal injection of GM6001, an MMP inhibitor, or TNF-α neutralizing antibody (anti-TNFα Ab) attenuated capillary degeneration in retinas of NMDA-injected eyes. Protein levels of TNF-α increased
2 h after NMDA injection, whereas those of MMP-9 increased 4 h after the injection. Anti-TNF-α Ab
suppressed activation of MMP-9 in retinas of NMDA-injected eyes, whereas GM6001 diminished the TNF-α
protein expression. Incubation of recombinant TNF-α with supernatants of homogenized retina increased
protein levels and activity of MMP-9. These results suggest that TNF-α and MMP-9 collaboratively
contributes to the progressive capillary degeneration in injured retinas.

Copyright © The Japanese Pharmacological Society4-B-P-275

一般演題（ポスター）

Transcription factor ERG controls endothelial cell function and vascular
normalization in tumor angiogenesis〇亀井 竣輔1、荒田 佳菜子1、宮村 優里1、荒木 喜美2、久保田 義顕3、南 敬1
1

熊本大・生命資セ・分子血管、2熊本大・生命資セ・疾患モデル、3慶應義塾大・医・解剖

Tumor microenvironment, in particular tumor angiogenesis, forms a niche that facilitates rapid tumor growthand metastasis. We have reported ERG, an ETS family transcription factor downregulated within the tumor
endothelial cells (ECs), is critical for increasing H3K27ac enrichment and mRNA transcription on EC-specificgenes (

2016,2018). However, the role of ERG in tumor angiogenesis is unclear.

Here we successfully generated EC-specific mouse ERG-transgenic mouse (iEC/Cdh5-Cre-ERT2;) andevaluated the tumor growth and angiogenesis in Lewis Lung Carcinoma (LLC)-transplantation model. We
observed differential angiogenesis patterns: a decrease in blood vessel number, a 1.5-fold increase in pericytes
coverage, and a 7 % expansion of blood vessel diameter within the tumor iniEC

mice. AndiEC

mice showeda 15 % decrease in tumor size in basal conditions, meanwhile, a significant 50 % decrease with a combination ofanti-cancer drug cisplatin-treated conditions. RNA-sequencing analysis additionally revealed differential gene
expression patterns related to inflammation, cell cycle, and cell growth on the smaller tumor tissues in thecisplatin-treatediEC

mice than in the cisplatin-treated control mice. These results suggested that thetranscriptional regulation of ERG was crucial for modulating EC function and the normalization of tumor bloodvessel environment via preventing ERG transcriptional decrease was a novel molecular target for developing
cancer drugs.

Copyright © The Japanese Pharmacological Society4-B-P-276

一般演題（ポスター）

Toxicity assessment of tyrosine kinase inhibitors using human-induced
pluripotent stem cell-derived cardiomyocytes〇林 紗代、佐塚 文乃、柳田 翔太、諫田 泰成
国立医薬品食品衛研・薬理部

BCR-ABL tyrosine kinase inhibitors (TKIs) have contributed to the improvement of the survival of patientswith chronic myelogenous leukemia (CML). Growing evidence suggests that cancer therapy-related cardiac
dysfunction has become important as the most undesirable side effect of chemotherapy. For example, one of
BCR-ABL TKIs, nilotinib, has been reported to have a risk of QT prolongation associated with Torsades desPointes and cardiac failure. We have previously reported that nilotinib causes QT prolongation and early
afterdepolarization (EAD) using human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs).
In this study, we evaluated the effect of BCR-ABL TKIs on contractility using hiPSC-CMs.
We used iCell Cardiomyocyte 2.0 (Cellular Dynamics International). To assess the chronic cardiotoxicity of theseveral BCR-ABL TKIs including nilotinib, we analyzed the contractility velocity recorded by motion vector
analysis (SI8000, Sony).
We found that treatment with BCR-ABL TKIs decreased beating rate in hiPSC-CMs in a concentrationdependent manner. In addition, nilotinib and other BCR-ABL TKIs decreased the relaxation velocities.
These results suggest that BCR-ABL TKIs affect contractility in hiPSC-CMs. We are planning to analyze theadverse drug event reporting database using BCR-ABL TKIs to conform the in vitro data.

Copyright © The Japanese Pharmacological Society4-B-P-277

一般演題（ポスター）

The mechanism for impairing vasorelaxation response enhancement byperivascular adipose tissue in metabolic syndrome〇籠田 智美1,2、懐 理紗1、麓（丸山） 加菜1、篠塚 和正1
1

武庫川女大・薬・薬理２、2武庫川女大・バイオサイエンス研

Sex differences in the modulatory function of arterial tone by perivascular adipose tissue (PVAT) are reported.
We demonstrated that sex difference in enhancing vasorelaxation response by PVAT in renal arteries differs
between metabolic syndrome (MetS) model strains, SHRSP.Z-/IzmDmcr rats (SPZF) and SHR/NDmcr-

cp rats (CP), at the same age. The underlying mechanism of this discrepancy was investigated.
Systolic blood pressure (sBP) was measured using a tail-cuff method. Ring preparations with and without PVAT
were made from male and female SPZF and CP renal arteries at 23 weeks of age. Vasorelaxation and mRNAtranscript levels in PVAT were examined using organ bath methods and quantitative real-time polymerase chain
reaction, respectively.
sBP was higher in SPZF than CP but lower in females than males. PVAT increased acetylcholine-induced
relaxations in renal arteries in CP females only. There were no significant differences in mRNA levels ofangiotensin II type 1 receptor (AT1R) and AT1R-associated protein (ATRAP) among groups, but the
AT1R/ATRAP ratio, which indicates AT1R activity, was lower in CP than SPZF. The AT1R/ATRAP ratio in
PVAT was negatively correlated with enhancing the effects of PVAT on acetylcholine-induced relaxations while
positively correlated with sBP.
This study suggests that overactivation of AT1R signaling in PVAT, probably resulting from high bloodpressure, induces the decline in compensatory PVAT effects in MetS. Negative regulation of AT1R signaling isbeneficial for sustained favorable PVAT effects.

Copyright © The Japanese Pharmacological Society4-B-P-278

一般演題（ポスター）

Administration of nicotinamide mononucleotide attenuates cardiotoxicity andskeletal muscle atrophy induced by doxorubicin in mice.

〇久野 篤史、細田 隆介、岩原 直敏、野島 伊世里
札幌医科大・医Doxorubicin (DOX), an anti-cancer drug, induces cardiotoxicity and skeletal muscle atrophy. We
recently reported that resveratrol, an activator of NAD+-dependent deacetylase SIRT1, prevented DOX-induced
cardiotoxicity. In this study, we examined effects of nicotinamide mononucleotide (NMN), a precursor of NAD+,on DOX-induced cardiotoxicity and skeletal muscle atrophy.
Male mice were randomly divided into three groups: vehicle, DOX, and NMN+Dox groups. In the
DOX group, mice received DOX (5 mg/kg, IP) once a week for four times. NMN (500 mg/kg, IP) was
administered to mice in the NMN+DOX group 30 min before and 2 days after DOX injection.
Echocardiography was performed at 1 week after the final DOX injection. Heart weight and tibialis anterior(TA) muscle weight were measured at 1 week after final DOX.
Body weight was gradually decreased in the DOX group compared with the vehicle group. NMN
partially blocked the decrease in body weight by DOX. Left ventricular fractional shortening (FS), an index ofcardiac contraction, was lower in the DOX group than the vehicle group but was preserved in the NMN+DOX
group. Heart weight-to-tibia length (TL) ratio and TA weight-to-TL ratio were lower in the DOX group,
indicating myocardial and skeletal muscle atrophy by DOX. NMN treatment blocked the reductions heart
weight and TA weight induced by DOX.
These findings suggest that supplementation of NAD+ by administration of NMN could be atherapeutic strategy for prevention of cardiotoxicity and skeletal muscle atrophy induced by DOX.

Copyright © The Japanese Pharmacological Society4-B-P-279

一般演題（ポスター）

A comparative study of the effects of cigarette smoke extracts (CSE) fromcombusted or heated cigarettes on contractile function, spontaneous beating2+
rate, and intracellular Ca dynamics in rat ventricular myocytes.

〇安田 純平1、松村 早希子1、陳 以珊1、納富 拓也1、堀之内 孝広2、西谷（中村） 友重1
和歌山県立医科大・医・薬理学講座、2北海道大・院医・細胞薬理学

1

Smoking is known as a risk factor for cardiovascular diseases. Recently, the use of heated cigarettes, which are
expected to be a less toxic, has increased in Japan. However, little is known about their direct effects on
cardiomyocytes. In the present study, we compared the effects of heated and a combustible cigarette smoke
extracts (CSEs) on the cell viability, contractile function and intracellular Ca2+ dynamics of freshly isolated or
cultured rat cardiomyocytes. We used Ploom X (PX) and IQOS (IQ) as heated CSEs, and 1R6F (RF) as a
combustible CSE. The cell viability (MTS) assay using cultured myocytes showed that the order of toxic effects
was RF>IQ>PX. Functional analysis using the Cell Motion Imaging System showed that all types of 1% CSE
reduced cardiomyocyte contractility with their impairing activity RF≒IQ>PX. Intracellular Ca2+ dynamics
analysis showed that all types of 1% CSE decreased the Ca2+-transient amplitude, which may be the mechanismof reduced contractility. In addition, RF treatment induce Ca2+ leakage (possibly from sarcoplasmic reticulum),which may be the cause of the increased diastolic Ca2+ levels. Furthermore, the spontaneous beating rate ofcultured cardiomyocytes was markedly reduced by RF. Taken together, these results indicate that heatedcigarette extracts are also toxic to cell viability and contraction of cardiomyocytes. In addition, each CSE had
different effects on intracellular Ca2+ dynamics, suggesting that mechanism of action is different.

Copyright © The Japanese Pharmacological Society4-B-P-280

一般演題（ポスター）

Inhibitory effects of eicosapentaenoic acid (EPA) on contractions in pig basilarand coronary arteries〇吉岡 健人、小原 圭将、追川 俊也、上村 洸平、山口 明奈、藤澤 和輝、花澤 瞳、藤原 実貴、遠藤 太尊、鈴木 太
智、De Dios Regadera Montserrat、伊藤 大地、斎藤 昂、中込 佑尚、山下 冬馬、木口 真由、齊藤 祐花、仲尾 友里、
宮路 陽南子、欧 光瀚、徐 可悦、田中 芳夫
東邦大・薬・薬理Eicosapentaenoic acid (EPA) is an n-3 polyunsaturated fatty acid (PUFA) found in fish oil. We recently showedthat docosahexaenoic acid (DHA), another n-3 PUFA, potently inhibited pig basilar and coronary artery
contractions induced by U46619 (a TP receptor agonist) and prostaglandin (PG) F2α. We also showed thatprostanoid TP receptors are potential targets for DHA. In this study, we investigated whether EPA, like DHA,
suppresses contractions of pig basilar and coronary arteries induced by U46619 and PGF2α through inhibition of
the TP receptor. EPA suppressed both U46619- and PGF2α-induced pig basilar and coronary contractions in aconcentration-dependent manner without affecting 80 mM KCl-induced contractions. U46619-/PGF2α-induced
contractions in both arteries were completely/largely suppressed by SQ 29,548 (a TP receptor antagonist). In
addition, EPA suppressed U46619-/PGF2α-induced increases in intracellular Ca2+ concentrations ([Ca2+]i) in
human TP receptor-overexpressing 293T cells, whereas it showed only a slight effect on PGF2α-induced [Ca2+]i
increases in human FP receptor-overexpressing 293T cells. These findings suggest that EPA strongly suppresses
TP-receptor-mediated contractions of pig basilar and coronary arteries, which can be partially attributed to its
inhibitory effects on the TP receptor.

Copyright © The Japanese Pharmacological Society4-B-P-281

一般演題（ポスター）

5-HT1A receptor activation induces oligodendrocyte transcription factors via theGab1/GSK3β signaling pathway in stress-maladaptive mice

〇黒川 和宏1、髙橋 浩平1、宮川 和也1、持田(斎藤) 淳美1、武田 弘志2、辻 稔1
国際医療福祉大・薬・薬理、2国際医療福祉大・福岡薬・薬理1

Our previous study demonstrated that 5-HT1A receptor activation by chronic administration of flesinoxan, a 5HT1A receptor agonist, reduces the abnormal emotionality in stress-maladaptive mice and promotes
oligodendrogenesis and myelination. However, the effects of 5-HT1A receptor activation on oligodendrocytetranscription factors under the stress-maladaptive situations and the underlying mechanisms remain unknown.
In the present study, we investigated whether activation of 5-HT1A receptor by flesinoxan could act onoligodendrocytes to induce oligodendrocyte transcription factors in stress-maladaptive mice. Western blot
analysis revealed that administration of flesinoxan significantly increased the expression levels of Gab1 and pGSK3β (Y216) in the hippocampus of stress-maladaptive mice, whereas decreased the expression level of pGSK3β (Ser 9) in the same brain regions. Under this condition, the significant increase in mRNA expression
levels of oligodendrocyte transcription factors, i.e. olig2, SOX10, MYF, NKX2.2 and Zfp24, in the hippocampus
were observed by real-time PCR analysis. The present findings suggest that 5-HT1A receptor activation may
induce oligodendrocyte transcription factors via the hippocampal Gab1/GSK3β signaling pathway and reduce
abnormal emotionality due to maladaptation to excessive stress.

Copyright © The Japanese Pharmacological Society4-B-P-282

一般演題（ポスター）

The critical role of PACAP/PAC1 signaling from the rhinal cortex to the
hippocampal astrocytes on the working memory〇神戸 悠輝、グエン トウ、栗原 崇、宮田 篤郎
鹿児島大・院医歯The neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) and its receptor (PAC1) are
widely distributed in the central nervous system. Although it has been reported that PACAP-PAC1 signaling
contributes to learning and memory, it is not clear what mechanism is involved in neuroplasticity. In this study,
we investigated the involvement of PACAP-PAC1 signaling in astrocytes for working memory. First, to confirm
the expression of PAC1 in hippocampal astrocytes, we obtained astrocyte-specific mRNA using the Ribo-tag
method, and RT-qPCR revealed that astrocytes expressed PAC1 six times higher than all cell types. A selective
knockout of hippocampal astrocytic PAC1 resulted in the impaired working memory of the Y-maze test. To
investigate the origin of PACAP neurons projecting to hippocampal astrocytes, we infected the hippocampus of
PACAP-Cre mice with adeno-associated virus (AAV), which is retrogradely transported and expresses mCherry
in a Cre-dependent manner, and observed mCherry-positive cells. We found some mCherry-positive cells in therhinal cortex. Furthermore, we infected the rhinal cortex of PACAP-cre mice with AAV, which expresses a
synaptophysin-EGFP chimeric protein cre-dependently. We found many synaptophysin-EGFP positive punctate
around hippocampal astrocytes, suggesting PACAP neurons in the rhinal cortex sent axons to the hippocampal
astrocytes. These findings suggested that PACAP from the rhinal cortex might act on hippocampal astrocytic
PAC1 and contribute to the working memory.

Copyright © The Japanese Pharmacological Society4-B-P-283

一般演題（ポスター）
2+

Analysis of in vivo astrocytic Ca activity in mouse models of neurodegenerative
diseases〇茂木 優貴、飯野 正光、金丸 和典
日本大・医・生理学分野

Astrocytic contribution to neurodegenerative diseases is attracting attention as a potential target for drug
discovery and therapeutics. Intracellular Ca2+ signaling in astrocytes is affected by bioactive
substances from damaged brain cells including neurons. This can trigger
theenhancement and/or attenuation of Ca2+-dependent processes in astrocytes, which lead to an alterationof gene expression profiles and secretion of neuroprotective and neurotoxic molecules. Thus, analysisof astrocytic Ca2+ activities may provide clues to regulatingneurodegenerative diseases. However, it remains
elusive how the pathological conditions affect astrocytic Ca2+ activities. Therefore, we are trying to establish a
new method to analyze astrocytic Ca2+ activities in neurodegenerative diseases. We applied drug-induced or
geneticmanipulation-induced neurodegenerative disease models to transgenic mouse linesexpressing a Ca2+

sensor protein or a Ca2+ signal-suppressing enzyme in astrocytes. Using macroscopic

Ca2+ imaging

and optical clearing method-assisted volumetric immunohistochemical analysis, wefound correlations between astrocytic Ca2+ activities and neurodegenerative phenotypes. These results andfurther analysis may contribute to the development of therapeutic strategies for neurodegenerative diseases.

Copyright © The Japanese Pharmacological Society4-B-P-284

一般演題（ポスター）

Impact of astrocytic lactate release on brain ischemic tolerance〇平山 友里1、小泉 修一2,3、安西 尚彦1
1

千葉大・院医・薬理、2山梨大・院医・薬理、3山梨大・院医・GLIAセンター

Brain ischemic tolerance is an endogenous neuroprotective mechanism, whereby an experience of non-lethal
ischemic episode (preconditioning; PC) produces resilience to subsequent lethal ischemia. We previously
showed that PC caused activation of astrocytes and a subsequent upregulation of P2X7 receptors (P2X7Rs),
activation of which induced ischemic tolerance. However, the downstream signals of P2X7Rs responsible for theischemic tolerance remain unknown. Here we show that astrocytic P2X7R-dependent lactate release has an
indispensable role for this event. Using a middle cerebral artery occlusion model, we found that extracellularlactate levels during lethal ischemia were significantly increased in mice that experienced PC, and the increasedepended on P2X7Rs. In theexperiments, although stimulation of astrocytes with P2X7R agonist

BzATP had no effect on the protein levels of MCT1 and MCT4, which were responsible for lactate efflux inastrocytes, BzATP induced the plasma membrane translocation of these MCTs via their chaperone CD147.
Importantly, CD147 was increased in activated astrocytes after PC, and CD147 blocking antibody abolishedastrocyte-mediated lactate release and ischemic tolerance. Taken together, our findings suggest that astrocytes
induce ischemic tolerance by P2X7R-dependent lactate release.

Copyright © The Japanese Pharmacological Society4-B-P-285

一般演題（ポスター）

Functional expression of choline transporters in microglia and their regulation of
microglial M1/M2 polarization〇稲津 正人1,3、岡田 寿郎2、武藤 瑛祐2、山中 力3、内野 博之2
東京医大・医総研、2東京医大・麻酔科学分野、3東京医大・分子予防医学

1

Microglia are key cells of the immune system in the central nervous system and are suggested to be deeply
involved in the development of neurodegenerative diseases. It is well known that microglia have functional
plasticity, with an inflammatory M1 phenotype and an anti-inflammatory M2 phenotype. Inhibition of cholinetransport in macrophages has been reported to suppress the secretion of inflammatory cytokines. However, therole of the choline transport system in regulating microglial M1/M2 polarization has not been fully elucidated todate. In this study, we investigated the mechanism of choline uptake in mouse microglial cell line SIM-A9, andits association with microglial M1/M2 polarization. Choline transporter-like protein 1 (CTL1) were highly
expressed in SIM-A9 cells and were localized in the plasma membrane. Functional analysis of choline uptake
demonstrated the existence of Na+-independent, pH-dependent, and intermediate-affinity transport systems.
Choline uptake was concentration-dependently inhibited by hemicholinium-3 (HC-3). Expression of the mRNAof M1 microglia markers IL-1β and IL-6 was increased by LPS, and their effects were suppressed by choline
deprivation and HC-3. In contrast, mRNA expression of the M2 microglial marker arginase-1 was increased by
IL-4, and the effect was enhanced by choline deprivation and HC-3. Our results suggest that inhibition of
CTL1-mediated choline uptake in microglia preferentially induces M2 microglia polarization, which is apotential therapeutic approach for inflammatory brain diseases.

Copyright © The Japanese Pharmacological Society4-B-P-286

一般演題（ポスター）

The astrocytic TRPA1 channel plays a protective role in vascular cognitive
impairment〇抱 将史1、中島 弘貴1、戸堀 翔太1、川下 綾香1、宮之原 遵1、森嶋 美沙1、永安 一樹1、繁冨 英治2,3、小泉 修一2,3、
森 泰生4、白川 久志1、金子 周司11

京都大・院薬、2山梨大・医・薬理、3山梨大・グリアセンター、4京都大・工・分子生物化学

Vascular cognitive impairment (VCI) is a syndrome defined as cognitive decline caused by vascular disease,which is associated with Alzheimer's disease and vascular dementia. Since chronic cerebral hypoperfusion(CCH) is commonly present in various types of dementia and induces VCI, we used bilateral common carotid
artery stenosis (BCAS) mice as a CCH-induced VCI model. Transient receptor potential ankyrin 1 (TRPA1),
the most redox-sensitive TRP channel, is functionally expressed in the brain and seems to function as apolymodal sensor in vascular disease. To clarify the involvement of TRPA1 in CCH-induced VCI, we usedgenetically engineered mice: TRPA1-knockout (TRPA1-KO) and cell-specific conditional TRPA1-KO mice. We
showed that TRPA1 deficiency exacerbated BCAS-induced cognitive impairment and white matter injury during
early-stage CCH. TRPA1 stimulation with cinnamaldehyde ameliorated BCAS-induced outcomes. We also
revealed that BCAS increased a cytokine in astrocytes. Moreover, TRPA1-stimulated primary astrocyte cultures
expressed the cytokine, and culture medium derived from these TRPA1-stimulated cells promoted
oligodendrocyte precursor cell myelination. Overall, TRPA1 stimulation in astrocytes plays a protective role in
CCH-induced VCI through the cytokine production.

Copyright © The Japanese Pharmacological Society4-B-P-287

一般演題（ポスター）

Expression and function of equilibrative nucleoside transporters in cultured
astrocytes〇田中 康一1,2,3、井澤 琢人1、冨田 和男1,2、五十嵐 健人1,2、北中 順惠3、北中 純一1、佐藤 友昭2、西山 信好1
1

兵庫医科大・薬・薬理、2鹿児島大・院医歯・歯科薬理、3兵庫医科大・医・薬理We have found that cultured differentiated astrocytes pretreated with

, - -dibutyryladenosine 3',5'-cyclic

monophosphate (DBcAMP), a permeable analogue of cAMP, incorporate thymidine, but not uridine, via
nucleoside transporters including equilibrative nucleoside transporters (ENTs） into TCA insoluble fraction forrepair on DNA injury in the presence of hydrogen peroxide (H2O2) at an early time, and these phenomena are
specific in differentiated astrocytes, but not undifferentiated astrocytes and neurons.
We studied expression and function of ENTs and LIMPII (Lysosomal Integral Membrane Protein II). We could
confirm ENT1, that is hypersensitive nucleoside transporter, and ENT2, that is low-sensitive nucleosidetransporter in cultured astrocytes by RT-PCR and western blot analysis and [3H]thymidine incorporation
experiment. Astrocytes were double-stained by anti-GFAP antibody and anti-ENT3 antibody. We could confirm
ENT3, that is assumed to be presented in lysosome, in cultured astrocytes co-stained by GFAP. We found theexpression of ENT3 and LIMPII by RT-PCR and western blot analysis, and the coexpression of ENT3 and
LIMPII in cultured astrocytes by immunocytochemistry. We clarified that ENT3 is localized in astrocytic
lysosome.
These results indicate that ENTs expressed and function in cell membrane and lysosome could relate with H2O2induced thymidine incorporation and DNA repair and disassembly in cultured astrocytes.

Copyright © The Japanese Pharmacological Society4-B-P-288

一般演題（ポスター）

Glial-derived secreted protein Tinagl1 regulates neuronal survival〇小椋 正人1、小野塚 星矢1、八巻 淳子1、和田 郁夫2、本間 美和子1
福島県立医科大・医・生体物質、2福島県立医科大・医・細胞科学

1Tubulointerstitial nephritis antigen-like 1 (Tinagl1), as a secreted matricellular protein, are implicated in themodulation of diverse processes including cell survival. We previously created transgenic mice expressing aphosphorylation-defective mutant of succinate dehydrogenase A (SDHAY215F) in astrocytes for evaluation of theeffects of mitochondrial ROS on astrocyte reactivity and showed that reactive astrocytes induce dopaminergic
neuronal loss though the expression of Tinagl1 in substantia nigra. In the present study, we investigate the effectof glial-derived Tinagl1 on neuronal survival. Conditioned medium prepared from primary astrocytes expressing
either SDHAY215F or Tinagl1 significantly decreased the number of microtubule associated protein 2-positive
cells as compared with control medium in primary neuron culture. Furthermore, recombinant Tinagl1(rTinagl1) also significantly induced neuronal apoptosis in a dose-dependent manner. Signaling analysis
revealed that rTinagl1 inhibits Akt activation, whereas promotes MKK4 activation in primary neuron culture.
These results suggest that glial-derived Tinagl1 may regulate neuronal survival through modulation of Akt and
MKK4 signaling. Exact molecular targets for Tinagl1 will be discussed.

Copyright © The Japanese Pharmacological Society4-B-P-289

一般演題（ポスター）

Proepileptic roles of astrocytes in mesial temporal lobe epilepsy withhippocampal sclerosis.

〇木下 慎一1、池谷 裕二1,2、小山 隆太1
1

東京大・院薬・薬品作用学教室、2東京大・Beyond AI研究推進機構

About 30% of patients with mesial temporal lobe epilepsy (mTLE), in which the hippocampus is the epileptic
focus, is drug-resistant to existing antiepileptic drugs that target the modulation of neuronal activity. mTLE iscommonly accompanied by the hippocampal sclerosis, which is primarily characterized by neuronal cell loss andchronic astrogliosis in hippocampal CA3 areas. It has been suggested that the changes in astrocytic function
associated with hippocampal sclerosis contribute to epileptogenesis, but the underlying cellular and molecularmechanisms remain unclear. Here, we found that during the development of hippocampal sclerosis, someastrocytes migrate from CA3 to the dentate gyrus (DG) and upregulate the gene expression that could modulate
neural activity related to the epileptic state. In intrahippocampal kainic acid (KA)-injected mTLE model mice,
we found that some astrocytes migrate from CA3 to DG during the early-stage of hippocampal sclerosis after
KA-injection. Then, we investigated the transcriptional profiles of the migrating astrocytes, finding an increased
expression of genes that could result in hyperactivation of neurons. Thus, our findings reveal previously
unknown changes in CA3 astrocytes during hippocampal sclerosis and propose astrocytes as a therapeutic target
for mTLE.

Copyright © The Japanese Pharmacological Society4-B-P-290

一般演題（ポスター）

HASTR/ci35, conditionally immortalized human astrocytes, can be a useful tool
for studies in characterization of human reactive astrocytes〇根岸 由佳、長谷川 理歩、山下 雅子、馬場 知代、森尾 花恵、降幡 知巳
東薬大・薬・個別化薬物治療学

Reactive astrocytes play important pathophysiological roles in various brain diseases in a context-dependent
manner, and they have thus gained significant attention as a target for development of new drugs. Therefore, we
aimed to characterize human astrocytes/conditionally immortalized, clone 35 (HASTR/ci35) as anreactive human astrocyte model. The results of quantitative PCR and RNA-sequencing showed that, upon
exposure to pro-inflammatory cytokines, HASTR/ci35 cells exhibited significant up-regulation of mRNA levelsof various inflammation associated genes, such as interleukin-6, intercellular adhesion molecule 1, and pentraxin
3. The results of the protein array confirmed their induction at protein levels. Qualitatively, the response profile
of HASTR/ci35 cells appeared to be similar to that observed in human primary astrocytes. To summarize,
HASTR/ci35 cells show an unequivocal reactive response to inflammatory stimuli, indicating that they can serve
as a human reactive astrocyte model. While further characterization is currently underway, it can be expectedthat they have a considerable potential to be used in reactive astrocyte-targeted drug development studies.

Copyright © The Japanese Pharmacological Society4-B-P-291

一般演題（ポスター）

Loss of the sustained antidepressant effect of (2R,6R)-hydroxynorketamine in
NMDA receptor GluN2D subunit knockout mice

〇山岸 愛実1,2、井手 聡一郎1、池窪 結子1、三品 昌美3、池田 和隆1
東京都医学総合研・依存性物質プロジェクト、2新潟大・院医歯・先端分子病態学、3立命館大・総合科学技術研究機構

1: The NMDA receptor antagonist ketamine exhibits acute and sustained antidepressant effects andis rapidly metabolized. We have reported that GluN2D plays an important role in the sustained antidepressanteffect of ( )-ketamine, an optical isomer of ketamine. Meanwhile, the pharmacological mechanism of major
metabolites of ketamine, ()-hydroxynorketamine (HNK), is still unclear. In this study, weinvestigated the role of GluN2D in the antidepressant effect of ()-HNK.

: We investigated the acute and sustained antidepressant effects of ()-HNK on 4 h restraint

stress-induced depression in wild-type and GluN2D-KO mice using the tail-suspension test (TST). After
behavioral tests, mouse brains were removed and brain regions considered to be related to the antidepressanteffect were collected and examined for protein expression of neuroplasticity-related molecules using theWestern Blotting (WB).
:()-HNK, but not ()-HNK, reduced immobility time in the TST 10 min after administrationin wild-type and GluN2D-KO mice. Further, the sustained antidepressant effect at 96 h post-treatment wasobserved only in wild-type mice and disappeared in GluN2D-KO mice. We analyzed the changes in protein
expression levels of molecules assumed to be related to neuroplasticity by the WB.
: Among the major metabolites of ketamine, ()-HNK has acute and sustained antidepressanteffects and GluN2D plays an important role in the sustained antidepressant effects of (Copyright © The Japanese Pharmacological Society)-HNK.

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一般演題（ポスター）

Resveratrol prevents dextran sulfate sodium-induced colitis-like symptoms anddepressive-like behavior by activation of AMP-activated protein kinase pathways
in the brain-gut〇高橋 浩平1、黒川 和宏1、洪 麗花1、宮川 和也1、持田（斎藤） 淳美1、武田 弘志2、辻 稔1
国際医療福祉大・薬・薬理、2国際医療福祉大・福岡薬・薬理1Patients with inflammatory bowel disease (IBD) have higher rates of psychiatric pathology including depression.
The dextran sulfate sodium (DSS)-treated mouse is a well-characterized animal model of colitis that exhibits
both IBD- and depressive-like symptoms. Recently, we found that DSS-treated mice decreased phosphorylationof AMP-activated protein kinase (AMPK) in the hippocampus and rectum. AMPK is known to be associatedwith the regulation of inflammation, autophagy, and neurogenesis. Hence, we examined whether AMPK
activator resveratrol (RSV) prevents the DSS-induced colitis and depressive-like behavior in mice. DSS-treatedmice exhibited the colitis-like symptoms, and depressive-like behavior in the tail-suspension test. Moreover,
DSS-treated mice showed the hippocampal and rectal decrease in p-AMPK and LC3Ⅱ/Ⅰ levels as well as
increase in p-p70S6K (a downstream target of mTORC1), p62, pro-inflammatory cytokines and cleaved caspase-3 levels, and the reduction of cell proliferation in the dentate gyrus of hippocampus. Treatment with RSV
prevented these abnormalities in DSS-treated mice, and induced LC3-positive puncta in the hippocampus.
These findings indicate that activation of AMPK in the hippocampus-gut may be involved in the antidepressantand anti-inflammatory effects of RSV in DSS-treated mice.

Copyright © The Japanese Pharmacological Society4-B-P-293

一般演題（ポスター）

Calcitonin gene-related peptide as an anxiety regulator in the mouse
hippocampus: Mechanisms for transcriptional regulation of monoamine oxidase
B by calcitonin gene-related peptide administration〇藤原 享志朗1、渡邉 杏香2、橋川 直也1,2、橋川 成美1,2
1

岡山理科大・大学院理学研究科・臨床生命科学専攻、2岡山理科大・理・臨床生命科学

Calcitonin gene-related peptide (CGRP) is a neuropeptide that affects anxiety; however, how it evokes anxietylike responses remains unclear. Here, we show that intracerebroventricular (i.c.v.) administration of CGRP (0.5nmol) increased monoamine oxidase B (MAOB) and decreased dopamine in the mouse hippocampus. CGRP
administration revealed anxiety-like behavior in open field, hole board, and plus-maze tests. CGRP decreased
dopamine but increased the transcriptional regulator of MAOB, Krüppel-like factor 11 (KLF11), and thephosphorylated heterochromatin protein (p-HP1γ), which is involved in gene silencing by methylating histone
H3 in mice hippocampus. Notably, increasing p-HP1γ becomes more euchromatic and activates transcription.
To determine whether this effect reflected the binding of HP1γ and KLF11 to the enhancer, we designed
primers within the KLF11 enhancer region (−700 bp) and performed chromatin immunoprecipitation (ChIP)
assays. Notably, HP1γ was recruited to the KLF11 enhancer by CGRP i.c.v. in the mouse hippocampus. We
also observed that CGRP (1000 pmol) infusion into the hippocampus significantly increased anxiety-likebehavior in the plus-maze test. Therefore, CGRP reduces hippocampal dopamine and exhibits anxiety-likebehavior through epigenetic regulation.

Copyright © The Japanese Pharmacological Society4-B-P-294

一般演題（ポスター）

Anxiolytic effects of inhibitors of endocannabinoid degrading enzyme on
anxiety-like behavior in restraint-stressed mice.

〇福森 良、中島 良佐、上尾 海南、山口 拓
長崎国際大・薬In this study, we investigated the effects of endocannabinoid (eCB) degrading enzyme inhibitors on anxiety-likebehavior and brain eCB levels in restraint-stressed mice.
For restraint stress, mice were forced in 50mL syringe for 30 minutes. Mouse brain was separated into theprefrontal cortex (PFC), hippocampus (HC), striatum (ST), periamygdaloid cortex (AM) and medulla
oblongata (MO). Brain tissues were homogenized with acetonitrile, which quantified for 2-arachidonoylglycerol(2-AG) or arachidonylethanolamide (AEA) content by UPLC/MS/MS. In addition, we measured anxiety levelsof mice in the elevated plus-maze test.
Restraint stress decreased 2-AG and AEA levels in PFC and HC, but not ST, AM, and MO. In the elevated plusmaze test, restrain-stressed mice showed anxiogenic behavior. This anxiogenic behavior was ameliorated byadministration of JZL184 (an inhibitor of monoacylglycerol lipase which hydrolyze 2-AG) or URB597 (aninhibitor of fatty acid amide hydrolase which hydrolyze AEA).
These results suggest that restraint stress induces anxiogenic behavior through the region-specific decrease ofeCB in PFC and HC. This anxiogenic behaviors were ameliorated by inhibitors of eCB degrading enzyme,
which indicates that anxiogenic behavior induced by restraint stress might be due to the reduction of eCB in
PFC and HC.

Copyright © The Japanese Pharmacological Society4-B-P-295

一般演題（ポスター）

Protective effects of antidepressants on amyloid β oligomers-induced
neurotoxicity in SH-SY5Y cells〇山本 謙1,3、辻 まゆみ2、小口 達敬1,2、門馬 佑太郎1,3、大橋 英朗1,3、野原 哲人1,3、井藤 尚仁1,3、永田 未希4、村上 秀
友3、木内 祐二1,2
1

昭和大・院医・薬理学部門 医科薬理学講座、2昭和大・薬理科学研究センター、3昭和大・医学部内科学講座・脳神経

内科学部門、4昭和大・薬・病院薬剤学講座[Introduction] It is widely known that patients with Alzheimerʻs disease (AD) often present depression, which isone of the risk factors for developing AD. Although antidepressants have been used in many patients with AD,
the evidence of their cognitive benefit remains inconsistent. AD is characterized pathologically by amyloidβpeptide (Aβ) deposition and neurofibrillary tangles in brain. In this study, we evaluated if treatment withantidepressants could modulate neuronal damage induced by the highly toxic Aβ oligomers (Aβo).
[Methods] Aβ1-42 aggregation was measured using thioflavin T. We prepared 5 μM Aβo solution using Aβ1
peptides and induced neuronal toxicity in SH-SY5Y cells (human neuroblastoma) using Aβo. SH-SY5Y cells-42

were treated with duloxetine (Dlx), venlafaxine (Ven), mirtazapine(Mir) and fluoxetine(Flx) and Aβo. Cell
viability and oxidative stress were measured.
[Results] Dlx and Flx inhibited Aβ1-42 aggregation in concentration-dependent manner. Dlx and Flx
significantly reduced Aβo-induced cytotoxicity compared to Aβo-alone group after 24 hr of treatment. All four
antidepressants reduced ROS production significantly in concentration-dependent manner compared to Aβ-oalone group after 30 min of treatment.
[Conclusion] These results suggest that antidepressants, especially Dlx and Flx, may be effective in preventing
and suppressing the progression of AD through multiple mechanisms such as inhibition ofand antioxidative effect.

Copyright © The Japanese Pharmacological SocietyAβ aggregation

4-B-P-296

一般演題（ポスター）

Involvement of the agranular insular cortex in antidepressant actions ofarketamine〇横山 玲1、吾郷 由希夫2、笠井 淳司1、田沼 将人1、林田 美鈴1、島﨑 雄人1、樋口 桃子1、五十嵐 久人1、勢力 薫1、山口 瞬3、中澤 敬信4、橋本 謙二5、橋本 均1,6,7,8,9
1

大阪大・院薬・神経薬理、2広島大・院医・細胞分子薬理、3岐阜大・院医・高次神経形態学、4東農大・生命科学、5千葉大・社会精神保健教育研究センター、6大阪大・院連合小児発達・子どものこころセンター、7大阪大・データビリティフロ
ンティア機構、8大阪大・先導的学際研究機構、9大阪大・院医・分子医薬( )-ketamine (esketamine), one of ketamine enantiomers, has been approved for treatment-resistantdepression. Although several preclinical studies have shown that another enantiomer ( )-ketamine(arketamine) has greater potency antidepressant-like effects than esketamine, the antidepressant mechanism ofketamine enantiomers is not fully understood. Here, we aimed to identify brain regions that contribute to thedifference in antidepressant action between ketamine enantiomers using isolation-reared mouse model ofdepression. We found that lower doses of arketamine than esketamine had antidepressant-like effects in
isolation-reared mice in the forced swim test. Then, the machine learning classifiers with brain-wide activation
mapping in isolation-reared Arc-dVenus reporter mice revealed that the agranular insular cortex (aIC) may
contribute to the antidepressant-like effect of arketamine and discrimination between the effects of ketamine
enantiomers. Furthermore, a temporary suppression of neuronal activity in the aIC blocked the antidepressantlike effect of arketamine, but not of esketamine, and conversely activation of neurons in the aIC induced
antidepressant-like effects in isolation-reared mice. These findings suggest that activation of the aIC is involved
at least partly in the antidepressant-like effects of arketamine.

Copyright © The Japanese Pharmacological Society4-B-P-297

一般演題（ポスター）

Evaluation of the effects of Kir4.1 channel inhibitors on lipopolysaccharideinduced depressive behavior〇清水 佐紀、中野 諒子、廣瀬 由佳、堀名 宏紀、大野 行弘
大阪医薬大・薬・薬品作用解析

Astroglial potassium channels (Kir4.1) are involved in the onset of depression. Previous study showed that
Kir4.1 channel is upregulated in the lateral habenula in rat models of depression (Nature, 554(7692), 323-327,
2018). In addition, it is reported that serotonin reuptake inhibitors (e.g., SSRIs) inhibit Kir4.1 channel currents,suggesting that astroglial Kir4.1 channels might be involved in the pharmacological action of antidepressants. In
this study, to explore the role of Kir4.1 channels in the treatment of depression, we evaluated the effects of theKir4.1 channel inhibitors, VU0134992 and quinacrine, on lipopolysaccharide (LPS)-induced depression in miceusing forced swim test. First, treatments of mice with VU0134992 (10‒30 mg/kg, s.c.) and quinacrine (10-100mg/kg, s.c.) reduced immobility time in despaired model by forced swimming. Next, we observed that LPS (0.8mg/kg, i.p.) increased immobility time in the forced swim test, and the immobility time was dose-dependently
reversed by quinacrine (30-100 mg/kg, s.c.). Especially, quinacrine (100 mg/kg, s.c.) significantly attenuatedprolonged immobility time by LPS in forced swim test. Furthermore, quinacrine (100 mg/kg, s.c.) alsosignificantly improved LPS-induced anxiety like behavior in the elevated plus maze test. These results
demonstrated that Kir4.1 channel inhibitors have antidepressant effects, suggesting that the Kir4.1 channel
inhibitors are useful as new antidepressants.

Copyright © The Japanese Pharmacological Society4-B-P-298

一般演題（ポスター）

Chronic administration of oxytocin exerts anxiolytic and antidepressant effects
in a dose-independent manner in corticosterone-induced depression model offemale mice

〇森 征慶、田村 美咲、隅 憲廣、川邉 隼輔、村田 雄介、大江 賢治、遠城寺 宗近
福岡大・薬・臨床薬物治療学

Oxytocin (OT) is thought to have potential as a new therapeuticstrategy for depression. In spite of the fact that women are at a far greaterrisk for depression than are men, many experimental studies use male subjects.
Thus, we investigated the anxiolytic and antidepressant effects of OT on
steroid-induced depression model of female mice.
Adult female C57BL/6J mice were injected with OT (0.01, 0.1, or
1.0 mg/kg, i.p.) and corticosterone (CORT; 40 mg/kg, s.c.) for 4 weeks. Mice
were randomly assigned to the following groups: (1) vehicle; (2) CORT; (3) OT
0.01 + CORT; (4) OT 0.1 + CORT; (5) OT 1.0 + CORT. To assess the anxiolytic andantidepressant effects of OT, mice were subjected to the open field test (OFT)and forced swimming test (FST).
OT 0.1 + CORT group showed a significantly higher number of
entries into the center zone in OFT than the CORT group. In the FST, theimmobility time of the OT (0.01, 0.1, and 1.0) + CORT groups were significantly
lower than that of the CORT group. The immobility time of OT (0.01 and 0.1) +CORT groups were comparable to the vehicle group.
Under CORT exposure, the anxiolytic effects of OT were seen onlyin the middle dose of OT (0.1 mg/kg), whereas OT treatment (0.01 and 0.1 mg/kg)
ameliorated the depressive behavior. Previous studies showed that OT (1.0mg/kg) improved both the CORT-induced anxiety and depressive behaviors inmales. These results suggest that the effective dose of OT may be different
between sexes.

Copyright © The Japanese Pharmacological Society4-B-P-299

一般演題（ポスター）

Regulation of proliferation by glutathione in neural/stem progenitor cells
generated after neuronal degeneration in the adult hippocampal dentate gyrus〇米山 雅紀、山口 太郎、尾中 勇祐
摂南大・薬Various neurological injuries are widely recognized as promoting endogenous neurogenesis in adult mammalianhippocampal dentate gyrus. Our previous studies demonstrated that the systemic treatment with trimethyltin
chloride (TMT) causes the granule cell loss in the dentate gyrus of adult mice, with being regenerated in thedentate granule cell after the neuronal loss. The goal of the present study was to elucidate the roles ofglutathione in proliferation of neural/stem progenitor cells (NPCs) after neuronal degeneration. Using the
NPCs isolated from the dentate gyrus of mice on day 3 post-TMT treatment, the exposure to buthionine
sulfoximine (BSO, inhibitor of glutathione synthesis) significantly attenuated the cell proliferation without cell
damage. Next, BSO-induced attenuation of proliferative activity was completely abolished by singly tested Nacethyl cysteine (NAC), which is precursor of glutathione. However, NAC did not affect the proliferative activityof the NPCs. Our results suggest that glutathione has a critical role in proliferative activity in the NPCs
generated following neuronal degeneration in the hippocampal dentate gyrus.

Copyright © The Japanese Pharmacological Society4-B-P-300

一般演題（ポスター）

The pivotal role of PACAP/PAC1 signaling in the locus coeruleus noradrenergic
system〇NGUYEN THI THU、栗原 崇、宮田 篤郎、神戸 悠輝
鹿児島大・院医歯・生体情報薬理学

Both pituitary adenylate cyclase-activating polypeptides (PACAP) and noradrenaline are known to be involvedin anxiety and fear memory. However, the crosstalk of these substances has not been clarified previously. Here
we investigated the PACAP action in the locus coeruleus (LC) noradrenergic system based on the mice. To
evaluate the significance of PACAP on the mice behavior, we used the PACAP-knockout (-/-) mice.
PACAP (-/-) mice less stayed in the open arm of the elevated plus maze than wild-type (WT) mice, suggesting
PACAP has an anxiogenic effect. When a fear conditioning test was conducted using PACAP (-/-) mice, thefreezing time was significantly decreased compared to WT mice suggesting that PACAP is important for fear
memory formation.
PAC1 is PACAP-specific receptor. To examine whether PAC1 was expressed in LC noradrenergic neurons, weobserved coexistence of tyrosine hydroxylase immunoreactivity and PAC1 mRNA, revealing expression of PAC1
in the LC noradrenergic neurons. To examine whether PAC1 functional expressed in the LC, we measured
extracellular noradrenaline in the hippocampus after PACAP treatment. Intracerebroventricular injection of
PACAP increased hippocampal noradrenaline, suggesting PACAP/PAC1 signaling in the LC is active.
From the results show above, PACAP/PAC1 signaling might activate LC noradrenergic system. In the future,
we will clarify the significance of LC PACAP/PAC1 signaling on mice behavior.

Copyright © The Japanese Pharmacological Society4-B-P-301

一般演題（ポスター）

Potential effects of Semaphorin3A-PlexinA signaling on the function andmetabolism of amyloid-beta precursor protein〇関口 拓己1、櫻井 隆1、山下 直也1,2
1

順天堂大・医・薬理、2神奈川工大・応用バイオ

Amyloid-beta (Aβ) aggregation has been believed to be the fundamental trigger of the development of
Alzheimerʻs disease (AD). Therefore, elucidating the mechanisms that induce Aβ overproduction from its type
I transmembrane precursor protein (APP) is one of the important issues in providing potential therapeutictargets for AD. Semaphorin3A (Sema3A), a secreted type of repulsive axon guidance molecule, is implicated in
the development of various neurodegenerative diseases. It was previously reported that Sema3A and its
signaling molecules accumulate and aggregate in AD patientsʻ brain. However, the molecular link between
Sema3A signaling and AD pathogenesis remains unknown. Here we show evidence that APP interacts with
PlexinA, a Sema3A receptor component. APP and PlexinA interacted through the extracellular regions and wewere able to narrow down these regions to less than 100 amino acids. Based on these findings, we are now
investigating whether the APP-PlexinA interaction affects APP function and metabolism, which might provide
new perspective that aberrant Sema3A signaling induces Aβ overproduction.

Copyright © The Japanese Pharmacological Society4-B-P-302

一般演題（ポスター）

In vivo molecular analysis of a novel gene involved in the maintenance of
wakefulness〇王 乙萌1,2、戸根 大輔1,2、山田 陸裕2、上田 泰己1,2
1

東京大・院医・システムズ薬理学、2理研・生命機能科学研究センター・合成生物学研究チーム

The molecular basis of mammalian sleep-wake regulation remains largely unexplored. Several studies haveidentified sleep-regulating kinases that strongly suggest that protein phosphorylation plays a key role inpromoting sleep (Tatsuki, 2016; Funato, 2016; Mikhail, 2017), but the regulation of protein phosphorylations
involved in the induction and/or maintenance of wakefulness remains unknown. In this study, we identified anovel gene involved in the maintenance of wakefulness. This geneX controls protein phosphorylation in various
cellular signaling pathways. AAV-mediated neuronal expression of gene X inhibits the transition fromwakefulness to sleep, leading to a marked increase in wake duration in mice.

We also found that this change inwake duration was accompanied by a significant decrease in delta power during NREM sleep. Conversely,
inhibition of ProteinX (encoded by geneX) function in neurons resulted in a decrease in wake duration. These
results imply that ProteinX is a key regulator of signaling involved in the maintenance of wakefulness. It has
been suggested that kinases such as CaMKII are activated during wakefulness and induce sleep, but in contrast,
ProteinX might exert its wake-promoting function through interaction with the sleep-promoting kinases.

Copyright © The Japanese Pharmacological Society4-B-P-303

一般演題（ポスター）

Acute effects of guanfacine on neuronal activities in the prefrontal cortex of mice

〇齋藤 文仁、鈴木 秀典、荒川 亮介
日本医科大・薬理学

Neurodevelopmental disorder, attention deficit hyperactivity disorder (ADHD) is considered to affect thewhole-brain, and one of candidate responsible brain region is the prefrontal cortex (PFC). The PFC regulates
high-order cognitive functions, including attention, behavior and planning through working memory. As a
medication for ADHD, α2A-adrenergic receptor agonist, guanfacine (GFC) has been shown to improve PFC
cognitive function, including working memory. However, it is poorly understood how GFC is effective in ADHD
pathology in the PFC. In the present study, we first investigated the acute effects of GFC on ion channels thatwere proposed to be modulated by cAMP-PKA pathway in brain slice preparation of C57BL/6 mice. We used
the somatic recording of patch-clamp techniques to record HCN channel- and KCNQ channel-mediatedcurrents from layer 5 neurons of the PFC (2~3 month-aged). However, GFC did not show obvious modulatory
effect on these current components. Next, we examined the modulatory effect of GFC on both EPSCs and
IPSCs onto pyramidal neurons. For the recording of synaptic currents, recorded neurons were classified to two
types (Callosal/Commissural (COM) and Corticopontine-projecting (CPn)). Although IPSCs were comparably
inhibited in both types of neurons, EPSCs were suppressed in only COM-type neurons. These results may beable to explain a part of therapeutic mechanisms of GFC. In the future, it is necessary to clarify whether thistarget cell-dependent modulatory action is involved in the clinical effect of chronic administration of GFC.

Copyright © The Japanese Pharmacological Society4-B-P-304

一般演題（ポスター）

Extracellular ATP-induced hyperexcitability: its molecular mechanism and its
role in synapse and behavior〇繁冨 英治1,2、鈴木 秀明1,2、平山 幸歩1、佐野 史和1,2,3、田中 謙二4、尾藤 晴彦5、小泉 修一1,2
山梨大院医・薬理、2山梨大院医・GLIAセンター、3山梨大院医・小児科、4慶應義塾大・医・先端医科研・脳科学、5東京

1

大院医・神経生化学

Astrocytes show dramatic changes at the molecular level in response to brain insult and disease, becoming
reactive astrocytes. Among the changes found in reactive astrocytes, we have focused on P2Y1 receptor(P2Y1R), which is activated by extracellular ATP and is upregulated in reactive astrocytes in some neurologicaldisorders including epilepsy. To reveal the pathophysiological significance of P2Y1R upregulation in astrocytes,
we have used transgenic mice in which astrocytes specifically overexpress P2Y1R using Tet-Off system(P2Y1OE). P2Y1OE mice were more susceptible to drug-induced seizures and showed more abnormal spikes in
EEG recordings, suggesting P2Y1OE triggered neuronal hyperexcitability. We analyzed the cellular mechanism
underlying the hyper-excitability by imaging techniques and electrophysiology in the hippocampal slices. We
found evidence showing that excitatory synaptic transmission was enhanced in P2Y1OE with the increase in
P2Y1R-mediated Ca2+ signals in astrocytes. Interestingly, the enhancement of excitatory synaptic transmission
was due to a novel excitatory molecule X derived from astrocytes rather than glutamate release from astrocytes.
Overall, our data show a novel mechanism of astrocytic regulation of excitatory synapses which could contributeto hyperexcitability in neurological diseases.

Copyright © The Japanese Pharmacological Society4-B-P-305

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Neocortical slow oscillations gate the transfer of hippocampal activity to thevisual cortex〇鮫島 華1、池谷 裕二1,2
1

東京大・薬・薬品作用学教室、2東京大・Beyond AI 研究推進機構

Recent evidence shows that the primary visual cortex (V1) is plastic and associated with storage of visual longterm memory. Although the hippocampal formation is not monosynaptically connected with the V1, neuronal
activity of the V1 is modulated during hippocampal ripples, a form of 150-250 Hz oscillations that plays a role inmemory consolidation, and the modulation may contribute to memory formation and retention; however, little is
known about subthreshold activity in individual V1 neurons during hippocampal ripples. We recorded
membrane potentials (Vms) from V1 layer Ⅱ/Ⅲ pyramidal cells of urethane-anesthetized mice using the wholecell current-clamp technique, together with recording local field potentials (LFPs) from the hippocampal CA1
region, to explore the Vm fluctuations in V1 neurons in response to hippocampal ripples. Time series analyses ofhippocampal LFPs and V1 neuronal Vm revealed a significant portion of V1 neurons are briefly depolarized
after the ripple onsets when they are at DOWN states of the Vm slow oscillations, whereas V1 neurons exhibiteda variety of responses (or no responses) after hippocampal ripples at the UP states. The latencies of thedepolarizations at DOWN states relative to the ripple onsets ranged from 20 to 40 ms, suggesting multisynaptictransmission from the hippocampus to the V1. Our findings advance our understanding about the neuronalmechanisms underlying neocortical consolidation of visually relevant memories.

Copyright © The Japanese Pharmacological Society4-B-P-306

一般演題（ポスター）

Oxytocin administration to head gamma-irradiated mice results in increased
KCC2 mRNA expression through decreased phosphorylated CREB

〇五十嵐 健人1,2、田中 康一1,2,3、北中 純一2、北中 順惠3、西山 信好2、富田 和男1,2、佐藤 友昭1
鹿児島大・院医歯、2兵庫医科大・薬、3兵庫医科大・医1

BACKGROUNDS: Cranial gamma-ray irradiation is one of the effective treatments for brain tumors. So far, wehave found that nasal administration of oxytocin prevents cognitive dysfunction in gamma-irradiated mice, andrestores reduction in mRNA expression of KCC2, a chloride ion transporter (Igarashi et al., BBRC, 2022).
However, it remains unclear how oxytocin is involved in the increased expression of KCC2.
METHODS: 3-week-old male C57BL6/J mice were exposed to a 60Co source for 4 minutes and irradiated with
1.5 Gy gamma rays. Irradiation was performed once a day and repeated for 3 days. Mice in the oxytocinadministered group were nasally administered 0.1 mM oxytocin 5 μL after each irradiation. Hippocampal tissue
was excised from gamma-irradiated mice, mice treated with oxytocin after gamma-irradiation, and non-gammaray irradiated mice, and the amounts of phosphorylated CREB and phosphorylated ERK1/2 were examined byWestern blot.
RESULTS: In mouse hippocampus administered nasally with oxytocin after gamma-irradiation, phosphorylated
CREB was reduced by approximately 24% compared with non-gamma-irradiated mice (p <0.05). In addition,
mice nasally administered oxytocin after gamma irradiation had a 51% decrease in phosphorylated CREB
(p<0.001) compared to non-irradiated mice, and a 35% decrease compared to gamma-irradiated mice
(p<0.05). In the mouse hippocampus to which oxytocin was nasally administered after gamma-irradiation, thephosphorylated ERK1/2 was increased by about 24% as compared with the non-gamma-ray-irradiated mouse,but it was not significant.
DISCUSSION: In this study, we found that phosphorylated CREB decreased in mice treated with oxytocin after
gamma irradiation. Previous studies have suggested that phosphorylated CREB is involved in the reduction of
KCC2 expression (Rivera et al., J. Neurosci. 2004). From these facts, it is considered that oxytocin
administration after gamma-ray irradiation may lead to an increase in the expression level of KCC2 through adecrease in phosphorylated CREB.

Copyright © The Japanese Pharmacological Society4-B-P-307

一般演題（ポスター）+

ATP-sensitive K channel activation in the brain is involved in elevation ofplasma levels of adrenaline〇岡田 尚志郎、山口 奈緒子
愛知医科大・医Recently, we reported that restraint stress-induced elevation of plasma levels of adrenaline, but not of
noradrenaline, was suppressed by intravenous and intracerebroventricular (i.c.v.) administration of glucose via
brain thromboxane A2-dependent mechanisms. (Yamaguchi et al., 2022). Since glucose can be converted to ATP
in the brain cells, in the present study, we investigate whether ATP-sensitive K+ channel (KATP channel) activity
in the brain is involved in changes of plasma levels of catecholamines in urethane anesthetized rats. I.c.v.
administered diazoxide (20 µg/rat), a KATP channel opener, and A-769662 (25 µg/rat), an AMPK activator,
elevated plasma levels of adrenaline, but not noradrenaline. In contrast, i.c.v administered glibenclamide (25µg/rat), a KATP channel antagonist, did not alter plasma levels of adrenaline. I.c.v. pretreatment with SQ29548(10 µg/rat), a thromboxane A2 receptor antagonist, attenuated diazoxide- and A-7969662-induced elevation ofplasma levels of adrenaline. These results suggest that KATP channel activation in the brain might trigger theactivation of thromboxane A2 receptor, resulting in elevation of plasma levels of adrenaline.

Copyright © The Japanese Pharmacological Society4-B-P-308

一般演題（ポスター）

Roles of sorting nexin protein 33 on amyloid precursor protein processing andthe level of amyloid β〇高鳥 悠記1、水川 裕美子1、漆谷 徹郎1、泉 安彦2、赤池 昭紀3、土田 勝晴1、尾﨑 惠一1、久米 利明4
同志社女子大・薬学部(京田辺)、2神戸薬科大・薬、3和歌山県立医科大・薬、4富山大・院医薬1

Alzheimer disease (AD), one of the most common diseases presenting dementia, is a progressive
neurodegenerative disorder characterized by the depletion of high-affinity nicotinic acetylcholine receptors(nAChRs) and a marked loss of cholinergic neurons. Selective depletion of cholinergic neurons in the basalis of
Meynert, decreased activities of choline acetyltransferase, or down-regulation of nAChRs suggest the roles ofcholinergic deficits in AD. Donepezil is a therapeutic acetylcholinesterase inhibitor currently being used for thetreatment of AD. However, recent studies have also reported off-target effects of donepezil that likely contributeto its therapeutic effects.We investigated the role of donepezil on the processing of amyloid precursor protein(APP) and the involvement of sorting nexin (SNX) 33, a member of the SNX protein family in this study.
Donepezil induced an increase in SNX33 expression. The expression of full-length APP in the cell lysate
remained unchanged, but the secretion of sAPPα in culture media increased. Donepezil led to a decrease inamyloid β (Aβ) protein levels in a concentration- and time-dependent manner. SNX33 knockdown by targetspecific morpholino oligos inhibited the effects of donepezil. Donepezil treatment increased cell membranesurface expression of APP in SNX33 expression-dependent manner. This study shows that donepezil increases
SNX33 expression and APP cleavage by α-secretase and reduces the level of Aβ by in primary corticalneurons.

Copyright © The Japanese Pharmacological Society4-B-P-309

一般演題（ポスター）

Neuroprotective effects of Raloxifene on Aβ oligomer-induced neuronal injury〇野原 哲人1,2、辻 まゆみ3、小口 達敬1,3、門馬 佑太郎1,2、大橋 英朗1,2、永田 未希4、井藤 尚仁1,2、山本 謙1,2、村上 秀
友2、木内 祐二1,3
1

昭和大・医・薬理学講座医科薬理学部門、2昭和大・医・内科学講座脳神経内科学部門、3昭和大・薬理学研究セン

ター、4昭和大・薬・病院薬剤学講座[Introduction] Increasing Alzheimer's disease (AD) is a social problem in an aging society. In addition, theprevalence of osteoporosis increases with aging and various interventions are being performed. The hypothesisthat aggregates and accumulation of Amyloid beta(Aβ) induces nerve cell death has been proposed. Raloxifene(Ral), a therapeutic agent for postmenopausal osteoporosis, is a selective estrogen receptor modifier, but it iscontroversial about slowing the progression of AD. Therefore, we investigated the protective effects of Ral on
Aβ-induced cytotoxicity.
[Method] The effect of Ral on the aggregation of Aβ was measured by the thioflavin T fluorescence method. In
addition, highly toxic Aβ42 oligomer (Aβo) was exposed to human neuroblastoma cells (SH-SY5Y) to induce
cytotoxicity. The effects of Ral on Aβo-induced cell damage were assessed by cell viability (MTT), oxidative
stress (ROS production, cell membrane phospholipid peroxide) and intracellular Ca2+.
[Results] Ral inhibited Aβ42 and Aβ40 aggregation in concentraiton-dependent manner. The effect of Ral on
the cytotoxicity of cells induced by Aβo exposure showed an increase in cell viability and a decrease in oxidative
stress, and repair of membrane damage. Ral reduced the sustained increase in intracellular Ca2+ due to Aβoexposure.
[Conclusion] Ral, a therapeutic agent for osteoporosis, has showed a protective effect against Aβo-induced
nerve damage by inhibiting Aβ aggregation and antioxidant effects. Ral is expected to have beneficial effects on
the prevention and progression of AD.

Copyright © The Japanese Pharmacological Society4-B-P-310

一般演題（ポスター）

Airway inflammation induced by LPS exposure in a mouse model of papaininduced asthma〇木村 元気、安藤 大稀、入江 孝祐、片山 侑紀、佐藤 しおり、鈴木 智大、原田 真衣、吉田 翼、西本 裕樹、木澤 靖
夫
日本大・薬In severe asthma patients, it is difficult to treat because of resistance to corticosteroid therapies. We havepreviously demonstrated that LPS induced steroid insensitive airway inflammation in mice. In this study, weinvestigated the effects of LPS exposure on airway inflammation in papain-induced asthma mice model. Papainsensitized A/J mice were challenged to papain every other day, then were exposed with LPS intranasally twice
daily for 3 days. Fluticasone propionate (FP) were administered intranasally at 2 h before each LPS exposure.
BALF was collected at 24 h after the last LPS exposure and eosinophils and neutrophils were quantified by
FACS analysis. The level of inflammatory cytokine and chemokine in BALF were measured by ELISA, andmRNA expression in lung was measured by RT-qPCR. The number of neutrophils and eosinophils in mice coexposed to papain and LPS in BALF was significantly increased compared to that in the papain alone-exposedgroup, especially the number of neutrophils was significantly increased. Chemokine and cytokine levels andmRNA expression were also significantly increased by co-exposure to papain and LPS, especially CXCL1.
However, these indicators of airway inflammation were resistant to FP. These profiles provide new insights into
steroid insensitive airway inflammation in severe asthma.

Copyright © The Japanese Pharmacological Society4-B-P-311

一般演題（ポスター）
ndDrug evaluation of LPS-induced acute lung injury (ARI) mice models (2 Report)〇牛島 壮太、和田 肇、岡本 公英、守住 孝輔、片山 誠一、廣中 直行、西 勝英(株)LSIM 安全科学研究所 熊本研究所・薬理研究部

In the 94th Annual Meeting of the Japanese Pharmacological Society, we reported two kinds of pulmonary
inﬂammatory models using lipopolysaccharide (LPS), LPS inhalation model and-GalCer-LPS instillation

model.
In the previous report, we concluded that these LPS-induced ARI models could be useful to evaluate therapeutic
efficacy of drugs used for treatment of pneumonia.
In the present study we carried out further research on LPS instillation model with a pretreatment of αgaractosyl ceramid (α-GalCer), and we also examined the effect of exosome from human adipose cell.
We evaluated inflammatory cell infiltration in BALF (Broncho Alveolar Lavage Fluid), BALF supernatantcytokine level and SpO2.
Exosome was found to be slightly effective to improve symptomatic parameters in this model. The action of
exosome changed along with the time of administration. Exosome administrated 4hr after LPS instillation
decreased body weight loss, inflammatory cell infiltration, IL-6 and IL-10 levels. On the other hand,administration of exosome 24hr after LPS instillation showed advanced histopathological appearance image ofthe lung.
The results indicate that characteristic pathophysiological change of this model was identified.

Copyright © The Japanese Pharmacological Society4-B-P-313

一般演題（ポスター）

Inhibition of androgen receptor signaling exacerbates pathological changes in amouse model of porcine pancreatic elastase-induced pulmonary emphysema.

〇青野 健太郎1、松本 純一2、松本 太一3、池田 弥恵2、河村 莉奈2、山内 淳史1
1

福岡大・院薬・生物薬剤学、2福岡大・薬・生物薬剤学、3福岡大・薬・医薬品情報学

Chronic obstructive pulmonary disease (COPD) is characterized progressive airflow obstruction due to thechronic inflammation in bronchitis and the alveolar destruction. We previously showed that testosterone
deficiency exacerbated porcine pancreatic elastase (PPE)-induced destruction of the alveolar structure inorchiectomy (ORX) mice, suggesting testosterone may have a suppressive effect on the progression of
pulmonary emphysema. However, it remains unclear the mechanism by which testosterone suppresses the
progression of pulmonary emphysema. In this study, we investigated whether testosterone supplementation
attenuates pathological change (body weight loss, the infiltration of T lymphocytes into the lung, and alveolar
destruction) in ORX/PPE mice. Furthermore, we examined effect of flutamide, androgen receptor (AR)inhibitor, on the enlargement of alveolar space in PPE-inhaled mice. Testosterone supplementation significantly
attenuated the loss of body weight, the infiltration of T cells in bronchoalveolar lavage fluid and alveolar
destruction in ORX/PPE mice. Furthermore, flutamide-treated mice show more severe emphysematous change
than vehicle-treated mice. These results indicate that testosterone-androgen receptor signaling have a pivotal
role in suppressing the progression of pulmonary emphysema through modulating T cell-mediated immune
response.

Copyright © The Japanese Pharmacological Society4-B-P-314

一般演題（ポスター）

Roles of the senescent alveolar epithelial cells in lung fibrosis using alveolar
epithelial cell-specific p16 knockout mice〇竹之内 康広、北風 圭介、杉本 理栄、石丸 浩靖、坪井 一人、岡本 安雄
川崎医科大・医Senescent cells are strongly implicated in various diseases including idiopathic pulmonary fibrosis. In ourprevious study, we confirmed expression of p16, a key marker of cellular senescence, in the alveolar epithelial
cells (AECs) of lungs from bleomycin-induced pulmonary fibrosis model mice. Although AECs have beenreported to contribute to lung fibrosis through epithelial-mesenchymal transition (EMT), it remains unclear
whether the induction of EMT is influenced by AEC senescence. In the present study, we investigated the rolesof AEC senescence in lung fibrosis using AEC-specific p16 knockout (cKO) mice. Intraperitoneal administrationof bleomycin caused pulmonary fibrosis in both control and cKO mice. However, the amount of soluble collagenin bronchoalveolar lavage fluid and collagen deposition in lung tissue, indicators of lung fibrosis, were lower incKO compared with control group. Furthermore, the mRNA expression of, which promotes lung fibrosis,

tended to be decreased in the lungs of cKO mice compared with controls. In addition, the expression of EMT
marker genes (and) also tended to be lower in the lungs of cKO mice. These results suggest that

cellular senescence mediated by p16 expression in AEC contributes to progression of lung fibrosis.

Copyright © The Japanese Pharmacological Society4-B-P-315

一般演題（ポスター）

Evaluation of insulin secretion by organ bath of isolated pancreases〇大内 基司1、森田 亜州華1、佐藤 慶太郎2、安西 尚彦1,3、藤田 朋恵1
獨協医大・医・薬理、2明海大・歯・薬理、3千葉大・院医・薬理1

In order to investigate molecules related to insulin secretion, we conducted a practical method to imitate humaninsulin secretion through rats via organ baths of pancreatic preparations. Our previous study showed that
insulin secretion from rat pancreas tissue was stimulated by glucagon-like peptide-1 and glimepiride. 1,5anhydro-D-glucitol (1,5-AG) is a glucose analog and exists in humans. This study aims to assess the effects ofshort-term 1,5-AG stimulation of insulin secretion in rat pancreases to better understand the effects in humans.
Rat pancreases were assigned to eight groups: two glucose concentrations (100 and 400 mg/dl) and pairs ofvarying 1,5-AG concentrations (0, 0.1, 1, and 10 mM). There was a significant increase in insulin outflow from
low to high glucose concentrations. However, there was no significant enhancement of insulin secretion between
the four groups with low and high 1,5-AG concentrations. This suggests that short-term exposure to 1,5-AG has
no effect on insulin secretion in rat pancreas tissues.
To justify whether the methods and techniques were useful as an experimental system, we isolated pancreases
from another rodent species, the mouse, and similarly measured insulin outflow. In mouse pancreas
preparations, stimulating with 400 mg/dl glucose significantly increased insulin outflow. Therefore, organ bathsof isolated mouse pancreases are also considerably effective for assessing effects of novel molecules and/or
therapeutics on insulin secretion.

Copyright © The Japanese Pharmacological Society4-B-P-316

一般演題（ポスター）

Mechanisms of the Antioxidant and Anti-inflammatory of SMTP-44D through Its
Soluble Epoxide Hydrolase Inhibitory Action〇篠内 良介1,2、柴田 佳太1,2、蓮見 惠司3、野部 浩司1,2
昭和大・薬・薬理、2昭和大・薬理科学研究セ、3東京農工大学・院農・応用生命化学専攻

1

We have previously reported the efficacy of SMTP-44D for diabetic neuropathy (DN) through its potential
antioxidant and anti-inflammatory activities. However, the mechanisms underlying the antioxidant and antiinflammatory activities of SMTP-44D remain unclear. The present study aimed to reveal the mechanism ofthese effects of soluble epoxide hydrolase (sEH) inhibition by SMTP-44D. In theassay, SMTP-44D (30mg/kg) was administered to 200 mg/kg streptozotocin (STZ)-induced diabetic mice from the 8 to the 28 days
after the injection of STZ. In theassay, IMS32 cells were incubated in a high glucose medium for 48 hand then treated with SMTP-44D (30 μM) for 48 h. The effects of the ratio of epoxyeicosatrienoic acids(EETs) to dihydroxyeicosatrienoic acids (DHETs), oxidative stress markers, and inflammatory factors byadministration of SMTP-44D were assessed by LC-MS/MS, TBARS, and ELISA assay, respectively.
Furthermore, apoptosis was evaluated by TUNEL assay. SMTP-44D treatment considerably increased the ratioof EETs to DHETs and mitigated oxidative stress, inflammation, and apoptosis. These results suggested that
SMTP-44D can suppress the induction of apoptosis by exerting antioxidant and anti-inflammatory effects,
possibly through sEH inhibition. SMTP-44D can be a potential therapeutic agent against DN.

Copyright © The Japanese Pharmacological Society4-B-P-317

一般演題（ポスター）

The influence of testes on diazepam-induced recognition memory impairment inrats〇畑 実野里1、石井 敦大2、川口 真以子1,2、畠山 太一3
1

明治大・農・動物環境学、2明治大・院農・動物環境学、3明治大・研究知財戦略機構

Benzodiazepines, including diazepam (DZP), induce recognition memory impairment as a side effect, and in
this class of drug, there are sex differences in pharmacokinetics and pharmacodynamics. Therefore, we focus onwhether the effect of DZP on recognition memory depends on sex difference. Our previous study showed theimprovement effects of ovaries on recognition memory deficit induced by DZP in female rats. Here we
examined the influence of testes on DZP-induced recognition memory impairment in male rats. Recognition
memory was assessed by a spontaneous object recognition (SOR) and a spontaneous place recognition (SPR)
tests that utilize the natural tendency of rats to explore a novel stimulus more than the familiar one. Thus, if rats
could retain the object identity or place information, they explored a novel object or place more than the familiar
one. First, using intact male rats, we examined the effect of DZP on SOR and SPR tests. Rats treated with DZP(2.0 mg/kg, i.p.) or vehicle, explored a novel object more than the familiar one in the SOR test, but in the SPR
test, they with DZP, but not vehicle, investigated equally two objects. Second, we had castrated intact male rats,
and then, the SOR and SPR tests were conducted. Results showed that in the SOR test, they with DZP or
vehicle explored a new object more than the familiar one, but in the SPR test, they with DZP, but not vehicle,
explored equally two objects. These results suggest that male rats even with DZP could retain the object
identity, but not place information, regardless of whether or not castration was conducted.

Copyright © The Japanese Pharmacological Society4-B-P-318

一般演題（ポスター）

Investigation of the function of DGKζ depending on its localization in pancreatic
β-cells〇渡邉 直也、金子 雪子、石川 智久
静岡県立大・院薬・薬理学講座

Increasing β-cell mass is a crucial therapeutic target for diabetes mellitus. Our previous study showed that
diacylglycerol kinase δ (DGKδ) is localized in the nucleus and acts as a suppressor of β-cell proliferation.
DGKζ, a type IV DGK isoform, has been shown to localize in the nucleus of rat β-cells and be a negative
regulator of cell cycle in the fibroblasts. We therefore examined the possibility that DGKζ, similarly to DGKδ,regulates β-cell mass. The intracellular localization of DGKζ was examined in MIN6 cells, a mouse β-cellline, and in isolated mouse islets by subcellular fractionation. Insulin secretion was measured by batch
incubation and cell cycle was analyzed by flow cytometry. In MIN6 cells, DGKζ was localized in the cytoplasm
and knockdown of DGKζ caused defective insulin secretion but did not affect cell cycle. In contrast, DGKζ inisolated islets was detected in the nucleus fraction as well as in the cytoplasm fraction. EGFP-tagged DGKζoverexpressed in MIN6 cells was also distributed in the nucleus. Moreover, overexpression of DGKζ induced
cell cycle arrest in MIN6 cells. These results suggest that cytoplasmic DGKζ regulates insulin secretion,
whereas DGKζ expressed in the nucleus regulates cell cycle. Thus, DGKζ is likely to have different function
depending on its localization in pancreatic β-cells.

Copyright © The Japanese Pharmacological Society4-B-P-319

一般演題（ポスター）

Intracellular serpin A1 regulates inflammatory cytokines expression via toll-likereceptor signaling pathway in endometrial stromal cells〇草間 和哉1、里吉 彩華1、安曇 麻奈1、吉江 幹浩1、梶原 健2、田村 和広1
東京薬科大・薬・内分泌薬理学、2埼玉医科大・医・産科婦人科学

1

Endometriosis is characterized by the presence of inflamed and fibrotic endometrial tissue outside the uterine
cavity. We have previously found that serpin A1 (SERPINA1) was decreased in the endometriosis-like lesion of
the mouse model, which makes us hypothesize that a decrease in SERPINA1 may exacerbate inflammation in
the lesion. However, the molecular mechanisms by which SERPINA1 affects the expression of inflammatory
cytokines and the development of endometriosis remains unclear. To investigate the role of SERPINA1 in
endometrial stromal cells (ESCs), RNA-seq analysis was performed using RNA extracted from ESCs knocking
down SERPINA1. The analysis has identified several toll-like receptor (TLR)-related factors as the upregulated
genes. Silencing SERPINA1 expression increased the expression of TLR3, TLR4, their downstream factor
MYD88, and inflammatory cytokines in cultured ESCs. Treatment with TLR3 or TLR4 agonists enhanced theexpression of inflammatory cytokines, whereas inhibitors of TLR3 or TLR4 decreased the expression of thesecytokines in SERPINA1-silenced ESCs. Immunohistochemical analysis showed that TLR3, TLR4, and MYD88
were localized in the endometriotic lesion. Thus, our data indicate that the decrease in SERPINA1 induces theexpression of inflammatory cytokines in ESCs, accompanying with TLR3/4 signaling. The regulation of
intracellular SERPINA1 could be a potential strategy to inhibit inflammatory responses in endometriotic lesions.

Copyright © The Japanese Pharmacological Society4-B-P-320

一般演題（ポスター）

Evaluation of anti-diabetic effects of the Cyclolepis genistoides D. Don (Paloazul), an herb native to Paraguay.

〇三竿 顕也1、北島 満里子2、村木 拓斗2、林 隼太郎2、高橋 晃輝1、福島 圭穣1、北井 淳一郎3、奥村 明子3、吉田 博
也3、石川 勇人2、藤野 裕道1
徳島大・院薬・生命薬理学、2千葉大・院薬・中分子化学、3株式会社 IHM

1

D. Don ("Palo azul" meaning "blue twig"; Palo) is an herb widely used for folk remedies
against diabetes and renal diseases in Paraguay.
The EtOH extract of Palo has been previously reported that it induced the mRNA expressions of peroxisome
proliferator-activated receptor γ (PPARγ) and adiponectin in mouse 3T3-L1 preadipocytes.
Here, to identify bioactive substances in Palo, a newly EtOH extracted crude sample (Palo-E) was separated byliquid-liquid extractions, and the AcOEt fraction (Palo-A), CHCl3 fraction (Palo-C), -BuOH fraction (Palo-B),
and water-soluble fraction (Palo-W) were obtained. 3T3-L1 cells were treated with each fraction 3 times in 7days and total RNA was collected followed by mRNA expression levels of PPARγ and adiponectin were
examined by RT-PCR.
As the results, treatment with the Palo-B and Palo-W fractions significantly increased mRNA expression of both
PPARγ and adiponectin in 3T3-L1 cells, but Palo-A fraction did not show these effects. On the other hand, cell
death was observed in the Palo-C fraction treated cells.
These results suggest that the Palo-B and Palo-W fractions are likely to contain bioactive substances, which
need to be identified by further separations in near future.

Copyright © The Japanese Pharmacological Society4-B-P-321

一般演題（ポスター）

Impact of chronic social stress on physiological and behavioral functions incynomolgus monkeys〇林田 健一郎1、堀本 泰弘1,2、稲留 大輔2、宮本 真二2、中村 祐里1、西方 龍太郎1,2、清水 洋志1、澤田 和俊1、沼田 洋
輔1、角崎 英志31

新日本科学・薬効薬理研究部、2新日本科学・実験動物管理部、3新日本科学・前臨床カンパニー

Chronic social stress (CSS) stemming from a low social position can result in depressive and anxiety disorders inanimals and humans. However, the pathophysiology of the CSS-associated disorders is still unclear. The present
study examined the effects of CSS on physiological and behavioral parameters in female cynomolgus monkeys,
by comparing those parameters in subordinates and dominants housed in pairs. In the blood, subordinates had
more white blood cells, fewer reticulocytes, and higher levels of low density lipoprotein than dominants.
Subordinates showed not only anxiety/depression-like behaviors but also impaired negative feedback control of
the hypothalamic-pituitary-adrenal axis and higher body temperature compared to dominants, consistent withsymptoms in patients with depression. In addition, magnetic resonance spectroscopy revealed that subordinates
showed less myo-inositol (an astroglial marker) in the amygdala compared to dominants, consistent withprevious observations in patients with depression. These results indicate that cynomolgus monkeys with CSS
present physiological and behavioral disorders that resemble the pathophysiology of depression in humans, andmay be a useful translational animal model for further research into the mechanisms and treatments ofdepression.

Copyright © The Japanese Pharmacological Society4-B-P-322

一般演題（ポスター）

Developing an experimental autoimmune encephalomyelitis (EAE) model incynomolgus monkeys: A feasibility study〇澤田 和俊1、中村 祐里1、西方 龍太郎1、深草 翔太1、清水 洋志1、林田 健一郎1、沼田 洋輔1、角﨑 英志2
1

新日本科学・薬効薬理研究部、2新日本科学・前臨床カンパニー

EAE is an animal model of multiple sclerosis (MS), which is most commonly used in rodents. Due to the recent
diversification of therapeutic modalities, including gene modification, and given the structural similarities in thecentral nervous system (CNS) to humans, non-human primates become to be considered a better animal model
for MS research. In the present study, we evaluated neurological signs, magnetic resonance images (MRI), andhistopathology in the CNS to examine whether immunization with myelin oligodendrocyte glycolipid (MOG)
could induce EAE in cynomolgus monkeys. After MOG treatment, the animals showed motor dysfunction andvisual impairment, indicating neurological deficits. However, there were individual differences in onset andseverity of the neurological deficits. MRI and histopathological examination revealed disseminated inflammationin brain and optic nerve, similar to the pathology of MS in humans. Although further research is required toreduce the individual differences of the neurological deficits, these results indicate that MOG treatment can
induce EAE in cynomolgus monkeys, and this can be used as a translational animal model for the developmentof new therapeutic modalities for MS.

Copyright © The Japanese Pharmacological Society4-B-P-323

一般演題（ポスター）

Examination of cognitive functions in MK-801 induced schizophrenia-likecynomolgus monkey model using Cambridge neuropsychological test
automated battery〇清水 洋志1、濱口 弘嗣1、大高 優1、西方 龍太郎1、中村 祐里1、澤田 和俊1、林田 健一郎1、沼田 洋輔1、角﨑 英志2
新日本科学・安全性研究所・薬効薬理研究室、2新日本科学・前臨床カンパニー

1

Cambridge neuropsychological test automated battery (CANTAB) is a cognitive function test device thatexamines memory, learning, attention, motivation, and reaction time in animals and humans. Schizophrenia is aserious mental disorder including positive and negative symptoms as well as cognitive dysfunction. In thepresent study, we utilized a delayed matching to sample task (DMST), one of the CANTAB programs, toexamine cognitive functions in a drug-induced schizophrenia-like cynomolgus monkey model. MK-801, an Nmethyl-D-aspartate (NMDA) receptor antagonist, that is known to induce schizophrenia-like symptoms invarious species, reduced accuracy rates in the DMST in the monkeys, demonstrating the cognitive impairment
from MK-801(20 μg/kg im, n=4). This effect was reversed by D-cycloserine, a partial NMDA agonist, andA776363, a dopamine 1 (D1) receptor agonist(0.1 and 1μg/kg im, n=3). On the other hand, donepezil(100μg/kg im, n=4), a cholinesterase inhibitor approved as an Alzheimer's disease therapeutic agent, did not affect
MK-801-induced cognitive impairment, similarly to the lack of donepezil efficacy on cognitive functions in
schizophrenia patients. These results demonstrate the roles of NMDA and D1 but not cholinergic receptors incognitive functions in MK-801 induced schizophrenia-like symptoms in cynomolgus monkeys.

Copyright © The Japanese Pharmacological Society4-B-P-324

一般演題（ポスター）

An early and versatile detection scheme of adverse drug reactions using largescale administrative claims data

〇山本 浩貴1、栢沼 玄1、長島 卓也2、戸田 千尋1、永安 一樹1、金子 周司1
京都大・院薬、2日本大・医1Adverse drug reactions (ADRs) are one of the leading causes of mortality and thus should be detected as early as
possible to reduce health risks of patients. Data mining approach using large-scale medical record might be auseful method for early detection of ADRs. There have been many researches analyzing medical record to detect
ADRs; however most of the researches focused only on a narrow range of late-onset ADRs, limiting its
usefulness. In this study, we developed an early and simple detection scheme of ADRs based on association rulemining (ARM) of JMDC insurance claims data. The assessment of its ability in detecting a broader range of
ADRs was performed using a global gold standard of ADRs consisting of 92 positive control ADR-drug pairs
and 88 negative control pairs rationally selected from the statistical analyses of large-scale ADR self-reports. In
this assessment of ARM, the areas under the receiver operating characteristic curve and the precision-recall
curve were 0.80 and 0.83, respectively. Moreover, when capability of ARM to detect ADRs with varying shortperiods of accumulating data was tested, our method detected much more positive controls (65 pairs) thanconventional sequence symmetry analysis (9 pairs) frequently-used for ADR detection. These results suggestthat ARM is effective in the early and versatile detection of ADRs, complementarily to the existing
pharmacovigilance strategies.

Copyright © The Japanese Pharmacological Society4-B-P-325

一般演題（ポスター）

A exploratory study on pharmacological actions of acetaminophen on ductus
arteriosus constriction based on a systematic review and database analysis
using Japanese Adverse Drug Event Report database〇石塚 洋一1、近藤 悠希1、難波 七海1、門脇 大介2
1

熊本大・大学院生命科学研究部(薬学教育)・臨床薬理学、2崇城大・薬・医療薬剤学

Background: Acetaminophen (APAP) is widely used as an antipyretic analgesic with a higher safety profile thannon-steroidal anti-inflammatory drugs (NSAIDs), however its pharmacological mechanisms are still fully
unclear. In this study, we conducted a meta-analysis and database analysis focusing on the effect of APAP on the"ductus arteriosus," i.e., the effect on treatment of patent ductus arteriosus (PDA) in low birthweight infants
and the occurrence of adverse reactions of fetal ductus arteriosus occlusion when administered to pregnant
women. Methods: Articles on the efficacy of treatment for PDA were collected, and a meta-analysis was
performed. The database analysis was conducted by collecting records about the fetal ductus arteriosus
contraction from the "JADER" database. Results and Discussion: The combined results of the randomized
controlled trials suggest that APAP has some therapeutic effect on PDA. The results of meta-analysis should beinterpreted with caution, given the insufficient number and quality of studies analyzed. In addition, an analysis
using JADER detected an adverse event signal of fetal ductal arteriosus occlusion was detected. At present,
these findings do not support the APAP therapy for the disease, nor rule out the use of APAP in pregnant
women. However, these data suggest that APAP shows the effects on ductus arteriosus.

Copyright © The Japanese Pharmacological Society4-B-P-326

一般演題（ポスター）
2+

Development of a functional antibody against Ca permeable channel TRPV2 fortreatment of human muscle degenerative diseases〇岩田 裕子、ピアソン ジェームズ
国立循環器病研究セ・心臓生理機能部

Abnormal Ca2+ handling is essential in the pathophysiology of muscular dysgenesis such as muscular dystrophy(MD) or/and dilated cardiomyopathy (DCM). One of Ca2+ permeable channels, transient receptor potential
cation channel, subfamily V, member 2 (TRPV2) has been suggested as a principal candidate for Ca2+ entry
pathways and a potential therapeutic target for muscular dysgenesis. Recently, we produced functional
antibodies against TRPV2 with remarkable protective effects against DCM as well as MD animal models.
However, the antibodies were rodent TRPV2 specific. Here, we produced selective antibodies recognizing
humanTRPV2 from the outside of cells using phage display. One of antibodies inhibited the Ca2+ influx via
humanTRPV2 in cultured cells and caused TRPV2 to disappear from the plasma membrane via cellularinternalization. We measured the change in [Ca2+]i induced by TRPV2 activator. Although cannabidiol induced
massively increased [Ca2+]i in the Duchenne muscular dystrophy (DMD) cell line, this increase was almost
completely abolished by treatment with the functional antibody. These results suggest that the functional
antibody we developed has therapeutic potential for treating MD or/and DCM.

Copyright © The Japanese Pharmacological Society4-B-P-327

一般演題（ポスター）

Microglia engulf myelin debris in mouse cortical slice cultures〇三上 弘記1、池谷 裕二1,2、小山 隆太1,2
1

東京大・院薬・薬品作用、2東京大・Beyond AI研究推進機構

Proper myelin formation by oligodendrocytes on neuronal axons is essential for brain function. Demyelination
causes various neurodegenerative diseases, but the cellular and molecular mechanisms underlying demyelination
remain unclear. Here, we developed a slice culture system to examine neuron-glia interactions during
demyelination.
To prepare slice cultures, postnatal-6-day mouse brains were sectioned, and then the cortical slices containing
the corpus callosum were cultured. By immunostaining cultured slices for the myelin marker Myelin basic
protein and the axonal marker Neurofilament, we confirmed that axons are myelinated in the corpus callosum
until days in vitro 15. Next, we attempted to induce demyelination in the slice culture system by applying
lysophosphatidylcholine (LPC), which is an endogenous lysophospholipid, from days in vitro 14 to days in vitro
15. LPC application induced a decrease in colocalization of Myelin basic protein and Neurofilament, which we
defined as demyelination, compared to the control group. Additionally, we found that microglia engulf myelin
debris around the demyelinated axons.
Thus, we developed a slice culture system in which demyelination and surrounding microglial response to
demyelination can be investigated.

Copyright © The Japanese Pharmacological Society4-B-P-328

一般演題（ポスター）

The standardization study to apply microphysiological systems (MPS) in drug
development as humanized pharmacological evaluation systems

最上（重本） 由香里1、北村（中山） 貴美子1、山崎 大樹1、石田 誠一1,2、佐藤 薫1
国衛研・薬理、2崇城大・院・工1

Microphysiological systems (MPS)s with human cells are attracting attentions all over the world as novel
pharmacological evaluation systems to improve human predictability of safety, PK/PD, and efficiency of new
drugs at the preclinical stage. In 2017, we started AMED-MPS project, the collaborative project among industry,
academia, and regulatory, in which we established the MPS prototypes of liver, small intestine, kidney, andblood brain barrier (BBB). However, the technical requirements for respective concepts of use (COU) andregulatory acceptance have not been studied sufficiently. We therefore have started the AMED MPS regulatory
science (RS) since 2Q of 2022 as the MPS standardization study. This is the first domestic national projectaiming at the development of commercially available MPSs ʼmade in Japanʻ. For the reference, we will show theongoing standardization research for BBB MPS. We have established the standard operation procedures (SOP)sto acquire the minimal essential benchmark data that show ʼBBB likelinessʻ. Lastly we summarized the global
trend and the domestic trend in MPS development based on our studies and previous reports.

Copyright © The Japanese Pharmacological Society