1-B-YIA01-1

一般演題(YIA)

Membrane proteome analysis for clarification of sex difference formation
mechanism of pharmacokinetics in the kidney〇清水 聡史1,2、橋口 丈晃1、Pornparn Kongpracha2、宮坂 政紀2、Pattama Wiriyasermkul2、坂本 多穗1、永森 收志2、黒川 洵子1
静岡県立大・薬・生体情報分子解析学、2東京慈恵会医科大・医・臨床検査医学

1

The kidney are the major organs determining the drug elimination, and membrane transporters play a central
role. There are reports that there are sex differences in the expression levels of transporters that are important indrug elimination function. However, the mechanism of sex difference formation is unclear. The one of reason isthat the low abundance and hydrophobic features of membrane proteins, such as transporters, make it difficult
to analyze the proteins. Therefore, we developed an original membrane proteomics method to analyze
membrane proteins comprehensively with high sensitivity. In this study, we used this method tocomprehensively analyze the membrane protein expression level of the Four Core Genotypes (FCG) model, in
which thegene was moved from the Y-chromosome to an autosome, allowing distinction between gonadal

and sex chromosome effects. Membrane proteomics was performed on brush border membrane vesicles from
FCG mice and obtained a membrane protein profile according to gonad and sex chromosomes. Next,
bioinformatics analyses were conducted to identify proteins and biological pathways that are important forgonad-derived and sex chromosome-derived sex differentiation. It is expected that this result will be an
important information on sex differences of drug elimination in the kidney.

Copyright © The Japanese Pharmacological Society1-B-YIA01-2

一般演題(YIA)

A pseudo-irreversible inhibition elicits a persistent effect of the sphingosine 1phosphate receptor-1 antagonist〇丸山 祐哉1,2,4、大澤 雄亮1、鈴木 孝幸1、山内 裕子1、大野 孝介1、井上 仁史1、山本 明俊1、林 守道1、宮内 真紀2,4、
広川 貴次3、秋山 泰身2,4
1

キッセイ薬品工業・中央研究所、2横浜市立大・院生命医科学・免疫生物学研究室、3筑波大・医学医療系、4理研・生命医科学研究センター・免疫恒常性研究チーム

Sphingosine 1-phosphate receptor-1 (S1P1), a G protein-coupled receptor, is required for the egress oflymphocytes from lymphoid organs into blood. Blocking S1P1 signaling is a promising therapeutic approach toinflammatory diseases, and the understanding of the structural basis for enhancing inhibitory activity is criticalto find potent S1P1 antagonists. We discovered a novel competitive antagonist, KSI-6666, that persistently
suppresses S1P1 signaling in vivo and effectively inhibit the pathogenesis in mouse colitis models. In silico
studies of molecular docking and molecular dynamics suggested that the dissociation of KSI-6666 from S1P1 isobstructed by the interaction of its bulky substituent with a methionine residue in the ligand-binding pocket of
S1P1. In vitro analysis revealed a pseudo-irreversible inhibition of S1P1 signaling by KSI-6666 and thestructural components that hindered its dissociation from receptors, corresponding to the predicted outcomesby molecular dynamics simulation. Moreover, in vivo studies suggested that the pseudo-irreversible inhibition
contributes to the persistent effect of KSI-6666. These findings would help the rationale design of potent S1P1
antagonists for the treatment of inflammatory disorders.

Copyright © The Japanese Pharmacological Society1-B-YIA01-3

一般演題(YIA)

Extracellular vesicles expressing brain-derived TrkB is detectable in serum: Itspotential as biomarker for cognitive enhancement〇山下 怜矢1、石本 尚大1、松本 聡2、増尾 友佑1、鈴木 誠2、加藤 将夫1
1

金沢大・薬学系、2エル・エスコーポレーション（株）

Brain-derived neurotrophic factor/TrkB signaling plays important roles in cognitive enhancement by promoting
neuroplasticity such as neurogenesis. Evaluation of the TrkB activation would be a potential biomarker forcognitive enhancement although its noninvasive evaluation is quite difficult. We hypothesized that TrkB would
be detected in serum extracellular vesicles (EVs) leaked from the brain, which would be a useful biomarker forcognitive enhancement. First, we checked secretion of TrkB-expressing EVs from the neuronal cells Neuro2atransfected with a plasmid encoding a flag tagged mouse TrkB (TrkB-flag). TrkB-flag was detected in EVs
isolated from the culture medium. Interestingly, intrahippocampal injection of adeno-associated virus serotype
PHP.eB vector encoding TrkB-flag gene (AAV-PHP.eB-mTrkB-flag) in mice showed that TrkB-flag was
detected in the serum EVs, implying secretion of TrkB-expressing EVs from the brain to circulating blood.
Finally, we analyzed the correlation between TrkB activation in serum EVs and cognitive enhancement in
humans administered ergothioneine (ERGO)-containing tablets, which are reported to activate TrkB in mice.
Oral administration of the ERGO tablets increased ratio of phosphorylated TrkB to TrkB, an indicator of TrkB
activation, in serum EVs, which was correlated with plasma ERGO levels and cognitive enhancement. Thesefindings suggest that TrkB-expressing EVs would be secreted into circulating blood, and its phosphorylated
states may help quantitative evaluation of cognitive enhancement.

Copyright © The Japanese Pharmacological Society1-B-YIA01-4

一般演題(YIA)

Exploring the novel role of mitochondria dynamics in neuron andoligodendrocyte differentiation〇岩田 圭子1、松﨑 秀夫2、野口 雅史1、新谷 紀人11

和歌山県立医大・薬・薬品作用、2福井大・子どもセ・脳機能発達研

Emerging evidence indicates that cell fate is pleiotropically regulated by mitochondria, which undergo specificdynamics (biogenesis, fission, fusion, and mitophagy) in the cells. In this study, we explored the possible roles ofthe mitochondrial dynamics (mtDYN) in neuronal and glial differentiation using

models. As models, weused human-derived cell lines SH-SY and MO3.13 cells, having the potential to differentiate into neurons andoligodendrocytes (OL), respectively. First, the transcriptome analysis on the mtDYN-related genes was
performed, and a common and marked increase was highlighted in the expression levels of peroxisome
proliferator-activated receptor gamma coactivator-1 alpha (PGC-1 alpha) in both differentiation models. The
differentiation-related increase of PGC-1 alpha, a key regulator of mitochondrial biogenesis, was also confirmedby an immunoblot study, and interestingly, mRNA analysis suggested the protein was produced from a noveltranscript variant, named OL-PCC-1 alpha. Further analyses in the OL differentiation model revealed that the
mitochondrial mass was dramatically increased, and moreover, specific knockdown of OL-PGC-1 alpha resulted
in a significant decrease of the mitochondrial mass and the expression OL differentiation marker proteins.
Collectively, the present data at least indicate that OL-PGC-1 alpha-related mitochondria biogenesis plays
promotive roles in OL differentiation, and open an avenue to study OL mtDYN in the pathophysiology
underlying brain dysfunction.

Copyright © The Japanese Pharmacological Society1-B-YIA01-5

一般演題(YIA)

Multistep regulation of mammalian sleep by phosphorylation states of CaMKII〇戸根 大輔1,2、大出 晃士1,2、張 千惠1、上田 泰己1,2
1

東京大・院医・システムズ薬理学、2理研・BDR・合成生物学

The sleep-wake cycle is an organism-level physiological phenomenon conserved in a wide range of species,
however, the molecular mechanisms remain largely unexplored. We have previously shown that Ca2+/calmodulin-dependent protein kinase II (CaMKII), one of the protein kinases regulated by intracellular Ca2+,

plays a critical role in promoting sleep (Tatsuki et al, 2016). CaMKII function is precisely controlled byphosphorylation in response to neural activity, suggesting that transitions of the CaMKII phosphorylation states
are deeply involved in driving the sleep-wake cycle. In this study, we comprehensively analyzed the effects of
CaMKII phosphomimetic mutations on sleep phenotypes in mice. We found that the phosphorylation of a single
residue in CaMKII could induce sleep from wakefulness. We further revealed that additional phosphorylation
switches the CaMKII function from sleep induction to sleep maintenance. These results provide evidence that
CaMKII plays multiple roles in sleep-wake regulation, depending on its phosphorylation state.

Copyright © The Japanese Pharmacological Society1-B-YIA02-1

一般演題(YIA)

Exploration of proliferation-promoting factors for human pancreatic beta cells〇前田 貴広1,2、木村 東2、伊藤 遼2、長船 健二21京都大・院医・医科学、2京都大・iPS研・増殖分化機構研究分野

Approaches that induce the proliferation of insulin-producing pancreatic beta cells hold promising therapeuticpotential to treat both insulin-dependent and insulin-independent diabetes. However, the factors that regulatebeta cell proliferation in human remain unknown, and drugs that control proper beta cell mass are in highdemand. Here, we aimed to identify small molecules that induce the proliferation of pancreatic beta cells as anovel therapy to treat diabetes. With this aim, we focused ongene, which is an essential driver ofproliferation in beta cells, and generated multiple human iPSC lines constitutively expressing a luciferase
reporter under the control of humanpromoter. Then, we developed a luciferase-based high-throughput

screening (HTS) system to detect small molecules that directly or indirectly enhance the activity ofpromoter in beta cells derived from the human iPSCs. Screens of 5,120 compounds using this system identified
several candidates with enhanced

promoter activity, and we confirmed that one compound promotes

human iPSC-derived beta cell proliferation. We are currently examining the mechanisms of action of thecompound. Our finding could contribute to the identification of novel targets for increasing beta cell mass.

Copyright © The Japanese Pharmacological Society1-B-YIA02-2

一般演題(YIA)

Investigation of Anti-Inflammation and Metabolome Shifts induced by SGLT2
Inhibitors to Elucidate the Mechanism of Cardioprotective Effects〇豊島 拓斗千葉大・院薬・臨床薬理学研究室

To investigate the possibility that Sodium-Glucose Cotransporter 2 Inhibitor (SGLT2i) reduces the risk ofcardiovascular events by suppressing inflammation in adipose tissue, this study examined the effects of SGLT2i
on inflammatory factors (IL-6, iNOS, MCP-1) in THP-1 derived macrophages and 3T3-L1 derived white
adipocyte cells to observe the effect of SGLT2i on inflammatory factors (IL-6, iNOS, and MCP-1). We also
observed the effect of SGLT2i on the comprehensive metabolome of white adipocytes: in THP-1, dapagliflozin(Dapa) and empagliflozin (Empa) showed a tendency to decrease IL-6 mRNA. In white adipocytes, 30 μM
Dapa significantly decreased IL-6 and iNOS mRNA and significantly increased methotionine (Met) and GABA.
Metabolomic pathway analysis also showed significant variation in Glu-related metabolism. Furthermore, Met
and GABA on white adipocytes tended to decrease IL-6 and MCP-1 in a concentration-dependent manner,
similar to SGLT2i. These findings suggest that SGLT2i may act on both adipocytes and macrophages. Futurestudies are expected to elucidate the mechanism of action of SGLT2i via Met and GABA, which were found tocorrelate with the suppression of inflammation.

Copyright © The Japanese Pharmacological Society1-B-YIA02-3

一般演題(YIA)

Branched-chain amino acid metabolism regulates gluconeogenesis via
mitochondrial pyruvate transport〇西 清人1、大野 美紀子1、西 英一郎1、Tian Rong2
1

滋賀医科大・薬理、2ワシントン大・医・麻酔

While it is well established that branched-chain amino acid (BCAA) catabolism is impaired in obesity, detailed
mechanisms how BCAA catabolism regulates glucose homeostasis is still elusive. While impaired BCAA
catabolism induces glucose intolerance in skeletal muscle or brown adipose tissues, the role of BCAAs and its
catabolism on liver metabolism is under-investigated.
The rate limiting step of BCAA catabolism is positively regulated by PPM1K through the dephosphrylation of
branched-chain keto acid dehydrogenase (BCKDH) complex. We found that Ppm1k-deficient (KO) mice, amodel of impaired BCAA catabolism, showed reduced gluconeogenesis and were protected from glucose
intolerance induced by high-fat diet feeding. In primary hepatocytes, accumulation of branched chain keto acids(BCKAs), downstream metabolites of BCAAs, due to Ppm1k deficiency inhibited hepatic glucose production ina cell-autonomous manner. Interestingly, pyruvate-supported glucose production was specifically suppressed in
KO mice or hepatocytes. Mechanistically, BCKAs directly inhibited liver mitochondrial pyruvate carrier (MPC)
activity resulting in selective suppression of pyruvate-supported gluconeogenesis or mitochondrial respiration.
Moreover, in non-hepatic mitochondria, BCKA accumulation was alleviated via the reversible reaction of
branched-chain amino transferase, rendering them less susceptible to BCKA-mediated inhibition of MPC.
Together, these results suggest that BCAA catabolism regulates pyruvate and energy homeostasis in liver via
BCKAs.

Copyright © The Japanese Pharmacological Society1-B-YIA02-4

一般演題(YIA)

Specific binding of uric acid to NDFIP1 associates with hyperuricemia-induced
liver fat accumulation〇朱 秋楠、荒川 大、谷口 愛美、長尾 優里佳、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

Uric acid (UA) is uniquely maintained at high concentration in higher primates. Contrastingly, hyperuricemia(HU) is a risk factor for gout and associates with diverse diseases, including NAFLD, while its mechanism
remains unclear. In this study, we aim to explain UA effect based on the hypothesis that UA-binding proteinsfunction as UA sensors and regulate various physiological processes.
To find UA sensors, pull-down assay using UA-binding beads and cell homogenate was carried out and 6
proteins were found as UA binding proteins. Specific binding of UA to these proteins was confirmed by binding
assay tosynthesized proteins, including NDFIP1. Meanwhile, we performed a proteomic analysis ofhuman primary hepatocytes treated with and without UA, followed by an enrichment analysis which showed
NAFLD is one of the remarkable diseases. When NDFIP1 was knocked down in HepG2 cells, an increase of fat
accumulation by UA exposure was attenuated, suggesting NDFIP1 plays a role in the HU-induced NAFLD.
Additionally, the proteomic analysis also showed UA exposure stabilized PTEN, whose ubiquitination is
promoted by NDFIP1. Furthermore, a decrease in PTEN ubiquitination and an increase in the protein level
were observed after UA exposure. Since the change of PTEN expression is reported to promote insulin
resistance that induces NAFLD, binding of UA to NDFIP1 is a possible mechanism of HU-induced NAFLD.
In conclusion, NDFIP1 was found as UA binding protein and was suggested as a UA sensor that regulates HUinduced NAFLD by modulating PTEN.

Copyright © The Japanese Pharmacological Society1-B-YIA02-5

一般演題(YIA)

Wnt5a, produced by mechanically stimulated periodontal ligament cells,
modulates differentiation of trigeminal ganglion cells〇高橋 かおり1、吉田 卓史2、中村 卓史1、若森 実1
東北大・院歯、2帝京平成大・薬・薬理1

The periodontal ligaments (PDLs), located between the tooth and alveolar bones, anchor the tooth and absorb
the daily chewing force by mastication. In addition, the Ruffiniʻs corpuscles in the PDLs play a pivotal role inpressure sensing to identify various food properties and to adjust occlusal force. Although recent studies haveshown that branches of Ruffiniʻs corpuscles are not formed without mechanical stimulation in the rat PDL(rPDL), there is no report to assess the regulatory mechanisms for the peripheral axonal structure bymechanically stimulated PDL cells.
We established primary PDL cell lines derived from rat molar tooth. The rPDL cells were loaded with periodic
mechanical stimulation (0.5 Hz, 15% expansion). The supernatant media of the mechanically stimulated PDL
cells enhanced neurite elongation, sprouting, and branching in trigeminal ganglion (TG) cells. Neurotrophic
factors such as NGF and BDNF and axon guidance proteins including Wnt family are known to be involved inneurite outgrowth. The rPDL cells expressed NGF, BDNF, NT-4 and Wnt5a mRNA. The mechanicalstimulation increased only Wnt5a in rPDL cells by the qPCR analysis and ELISA. Moreover, AP7677a(neutralizing anti-Ryk antibody), or strictinin (Ror1 inhibitor) suppressed the morphological changes. Thesefindings indicate the indirect mechanisms where Wnt5a, released from the connective tissues in response tomechanical stimulation, enhances the outgrowth of the peripheral nerves.

Copyright © The Japanese Pharmacological Society1-B-YIA02-6

一般演題(YIA)

Involvement of Pgc-1α in the skeletal muscle on glucose uptake via theperipheral sympathetic nervous system in rats

〇佐藤 大介1、今泉 希1、楠 正隆2、宮本 理人3
山形大・院理工・化学・バイオ工学、2名古屋大・総合保体セ、3神奈川工大・健康医療・管理栄養学

1

It has been suggested that sympathetic nerve activity is involved with not only blood pressure control but alsoenhancement of peripheral glucose uptake. To evaluate the effects of peripheral sympathetic activation on
glucose uptake and skeletal muscle mRNA expression involved with energy metabolism, in the present study, we
detected sympathetic nervous signal with a microelectrode in the unilateral sciatic nerve under anesthetic
condition in normal and high-fat diet-fed (HFD) rats, and applied electrical microstimulation (MS) via themicroelectrode. Glucose uptake was assessed as glucose infusion rate (GIR) during the hyperinsulinemiceuglycemic clamp. The MS significantly increased the GIR in both groups (P<0.01) whereas we observed nosignificant change in plasma insulin level. Furthermore, the GIR in HFD group was lower than that in theexpressions in the bilateral soleus

normal group. As a result of the MS for 60 min, in the normal group,and extensor digitorum longus (EDL) tended to be elevated while

expression in the bilateral EDL wassignificantly suppressed (P<0.05). In contrast, these changes were not seen in the HFD group. These results
suggest that the MS enhances non-insulin-mediated glucose uptake, and the effects may be mediated by several
pathways includingin the soleus but not in the EDL.

Copyright © The Japanese Pharmacological Society1-B-YIA03-1

一般演題(YIA)

15-hydroxyeicosatrienoic acid increases vascular permeability in nasal mucosa〇尾崎 乃理子1,2、堀上 大貴1、平山 和宏2、村田 幸久1,3
1

東京大・農・農学生命科学研究科 放射線動物科学研究室、2東京大・農・農学生命科学研究科 獣医公衆衛生学研究室、3東京大・農・農学生命科学研究科 食と動物のシステム学

Upon allergic rhinitis (AR), various inflammatory mediators increase blood flow and vascular permeability innasal mucosa, which result in nasal congestion.
We here investigated the effects of 15-hydroxyeicosatrienoic acid (15-HETrE) on functional changes ofvasculature and nasal congestion in mice. In isolated mouse aorta, the treatment of 0.1-3 µM 15-HETrE itself
did not induce any contraction but inhibited the TP agonist (U46619)-induced contraction in a dose-dependent
manner. Several lipid mediators are known to cause vasodilation by activating K+ channels. Consistently, a pretreatment of BKCa/IKCa, KV, or KATP channel inhibitor inhibited the 15-HETrE-induced relaxation. In vivo, thetopical administration of 1 µg U46619 constricted vein in mouse ear. A pre-administration of 1 µg 15-HETrE
also inhibited the U46619-induced vein constriction. In modified Mileʻs assay, an intranasal administration of 20µg 15-HETrE increased dye extravasation in the nasal mucosa. This administration also induced abdominal
breathing, immobility, and lying down, which can be caused by nasal congestion. Thus, 15-HETrE induced
vasorelaxation and vascular hyperpermeability. These phenomena presumably contribute to nasal congestion.

Copyright © The Japanese Pharmacological Society1-B-YIA03-2

一般演題(YIA)

2, 5-Dimethylcelecoxib induces accumulation of anti-inflammatory macrophages
and attenuates cardiac fibrosis in a mouse model of cryoinjury-induced
myocardial infarction〇岸上 赳大、石兼 真、有岡 将基、高橋 富美
産業医科大・薬理学講座

We previously reported that 2, 5-dimethylcelecoxib (DMC), a derivative of celecoxib, suppresses cardiac
remodeling by suppression of fibroblast-myofibroblast transformation. However, its effects on the
immunoreactive responses remain unclear. As macrophages are known to play critical roles in the process offibrosis after myocardial damage, we evaluated the effect of DMC on macrophages using a cryoinjury-induced
myocardial infarction (CMI) model mouse. The anterior left ventricular was cryo-injured by a liquid nitrogencooled aluminum probe in male C57 BL/6 mice. The mice were provided feed containing DMC or vehicle
starting 3 days before the operation. Echocardiography showed that DMC attenuated the impairment of cardiac
function, and Massonʻs trichrome staining of cross-section heart showed DMC reduced fibrosis area at the 14
days post-operation. In the cryo-injured damage area, DMC increased CD163-positive anti-inflammatory (M2)
macrophages 3 days after operation, but not CD86-positive pro-inflammatory (M1) macrophages. Real-time
PCR showed that DMC suppressed mRNA expression of interleukin (IL) -1β, IL-6, and monocyte
chemoattractant protein (MCP) -1, which are known as inflammatory cytokines, in the damaged myocardium.
These results suggested that DMC could attenuate impairment of cardiac function and fibrosis after the cardiac
damage through increasing accumulation of M2 macrophages and decreasing inflammatory cytokines. Thus,
DMC has potential against cardiac fibrosis and could be useful for the treatment of cardiac remodeling.

Copyright © The Japanese Pharmacological Society1-B-YIA03-3

一般演題(YIA)

β-arrestin-biased AT1R agonist improves health- and lifespan in mice withcongenital dilated cardiomyopathy〇川岸 裕幸1,2、Ramadhiani Risa3、冨田（沼賀） 拓郎2、中田 勉4、江本 憲昭3、山田 充彦2
信州大・バイオメディカル研・バイオテクノロジー、2信州大・医・分子薬理、3神戸薬科大・薬・臨床薬学、4信州大・基盤

1

共通支援セ・機器分析

Pediatric heart failure is an important cause of death in childhood; however, therapeutic drugs specific for
pediatric heart failure have not been developed. Angiotensin II modulates cellular function by associating with
its receptor, mainly angiotensin II type 1 receptor (AT1R) and activates G protein or β-arrestin signaling. We
reported that a β-arrestin-biased AT1R agonist (BBA) peptide, TRV027 induced a strong inotropic effect on
preweaning mice suffering from congenital dilated cardiomyopathy (DCM). Remarkably, this inotropic effectwas not associated with either arrhythmia or an increase in cardiac oxygen consumption. Here, we examined theprosurvival effect of TRV027 in DCM model mice. Daily subcutaneous administration of TRV027, but not an
AT1R blocker from postnatal day 1 significantly improved the survival rate and contractility of the left ventricle.
Hematologic and pathological analyses revealed no detectable abnormalities in TRV027-treated DCM model
mice except for hypertrophic heart. These results suggested that TRV027 has a beneficial effect to prolong
healthy lifespan of pediatric heart failure patients. We also performed a high-throughput screening of one
million compounds in order to discover small molecule BBA and found some hit compounds.

Copyright © The Japanese Pharmacological Society1-B-YIA03-4

一般演題(YIA)

TMEM182 maintains the activated state of Wnt/β-catenin signaling by increasing
ILK and inhibits cardiac differentiation in human iPS cells〇森原 啓文1、若林 繁夫1,2、朝日 通雄11

大阪医科薬科大・医・薬理、2大阪青山大・健康科学・看護

Transmembrane protein 182 (TMEM182) is known to be specifically expressed in muscle and adipose tissue.
Previous reports suggest that TMEM182 plays an important role in skeletal muscle development. However, therole of TMEM182 in cardiac muscle is still unknown, and this study aims to elucidate the role of TMEM182 in
the process of cardiomyogenesis. Human induced pluripotent stem cells (hiPSCs) can be converted intofunctional cardiomyocytes along the mesoderm lineage. Therefore, we generated hiPSCs overexpressing
TMEM182 in a doxycycline-inducible manner and induced differentiation into cardiomyocytes. At day12 ofdifferentiation, the expression of cardiomyocyte markers TNNT2 and MYH6 was significantly decreased in
TMEM182-overexpressing cells. Wnt/β-catenin signaling is activated during early differentiation andrepressed after mesoderm formation during cardiac differentiation. Therefore, we investigated Wnt/β-cateninsignaling during TMEM182 overexpression. We found that phosphorylation of GSK-3β (Ser9) and β-catenin(Ser552) was increased during TMEM182 overexpression, suggesting activation of Wnt/β-catenin signaling.
We further focused on integrin-linked kinase (ILK) as mechanisms by which TMEM182 activates Wnt/βcatenin signaling. The results showed that ILK expression was increased in cells overexpressing TMEM182.
These results suggest that TMEM182 maintains Wnt/β-catenin signaling activation after mesoderm formation
by increasing ILK expression and suppresses hiPSCs differentiation into cardiomyocytes.

Copyright © The Japanese Pharmacological Society1-B-YIA03-5

一般演題(YIA)

Nucleotide metabolism as a novel and potential target to regulate cardiomyocyte
proliferation〇齋藤 祐一、木村 航
理研・生命機能科学研究センター・心臓再生研究チーム

Heart failure is a life-threatening disease with a poor prognosis. The most common cause of heart failure is
myocardial infarction, in which a large number of cardiomyocytes are lost by occlusion of coronary blood flow.
In adult mammals, injured hearts cannot regenerate themselves because of the lack of proliferative capacity incardiomyocytes. In mice, cardiomyocytes permanently exit the cell cycle within 2 weeks after birth. Therefore,
understanding what molecular mechanisms underlying this permanent cell cycle arrest would provide animportant clue to identify potential therapeutic targets to induce heart regeneration. By integrated analysis of
metabolome and transcriptome, we found a drastic increase in nucleotide metabolism in the mouse heart during
this postnatal cell cycle withdrawal. Pharmacological inhibition of nucleotide metabolism in neonatal miceextended postnatal cardiomyocyte proliferation window with reduced oxidative DNA damage. Our findings
suggest that the nucleotide metabolism is a novel regulator of postnatal cardiomyocyte cell cycle.

Copyright © The Japanese Pharmacological Society1-B-YIA04-1

一般演題(YIA)

Effects of dexamethasone on nasal allergic response and hyperresponsivenessinduced by Japanese cedar pollen in mice

〇星野 楓月1、山下 道生2、安藤 祐介3、笠井 菜穂子1、田中 淑媛1、牧野 春香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

In the present study, effects of dexamethasone (Dex) 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, the sensitized mice were challenged by intranasal (Animals were also treated with Dex (.) 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. On days 18 and 25, the histamine (Hist,.)-induced sneezing was alsocounted. 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. 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 in sneezing. 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 pretreatments with
Dex. Thus, the murine AR model used might be useful for making clear the mechanisms of theAR pathogenesis and the action of corticosteroid effects.

Copyright © The Japanese Pharmacological Society.

1-B-YIA04-2

一般演題(YIA)

Protective effect of VEGFR1 signaling on LPS-induced lung injury in mice

〇長田 真由子1、伊藤 義也1、山下 敦1、田邉 美奈1、古江 明子1、細野 加奈子1、畑中 公1、澁谷 正史2、馬嶋 正隆1,3、
天野 英樹1
1

北里大・医・薬理、2上武大・医学生理学研究所、3神奈川工科大・健康医療・病態治療

Acute respiratory distress syndrome (ARDS) is characterized by increased permeability of pulmonary bloodvessels, leading to respiratory failure. Although VEGF is responsible for the vascular permeability, it remains
unknown about the involvement of signaling for VEGFR1, a receptor for VEGF. We examined the role of
VEGFR1 in pathology of ARDS. ARDS was created by an intra-tracheal injection of LPS to wild type (WT)
mice and VEGFR1 tyrosine kinase deficient mice (TKKO). Compared with WT mice, TKKO mice displayed
lower survival rates, increases in lung injury score, total protein concentrations, and pro-inflammatory cytokines
including TNF and IL-6 in bronchial alveolar lavage fluids. Alveolar macrophages were diminished in both typesof mice after LPS injection. Instead, neutrophils were extensively accumulated, and the number of neutrophils
in TKKO mice was higher than that in WT mice. The same was true for macrophages recruited into the lungtissues. VEGFR1 was expressed in alveolar and recruited macrophages. These results suggested that VEGFR1
signaling attenuated LPS-induced acute lung injury by suppressing vascular permeability, cytokines production
and neutrophil accumulation.

Copyright © The Japanese Pharmacological Society1-B-YIA04-3

一般演題(YIA)

ACE2-like carboxypeptidase B38-CAP suppresses severe acute lung injury
induced by aspiration pneumonia and abdominal sepsis as well as SARS-CoV-2
infection〇山口 智和1、湊 隆文1、星崎 みどり2、韮澤 悟3、安 健博1、高橋 砂織4、今井 由美子2、久場 敬司1
1

秋田大・院医・分子機能学・代謝機能学、2医薬基盤・健康・栄養研感染病態制御、3国際農研生物資源・利用領域、4秋

田県総合研

Angiotensin-converting enzyme 2 (ACE2) is the carboxypeptidase to degrade angiotensin II (Ang II) toangiotensin 1‒7 (Ang 1‒7) and improves the pathologies of cardiovascular disease and acute respiratory distresssyndrome (ARDS) / acute lung injury. B38-CAP is a bacteria-derived ACE2-like carboxypeptidase as potent as
human ACE2, recombinant B38-CAP is prepared withprotein expression system more efficiently thanrecombinant soluble human ACE2. We have demonstrated that B38-CAP ameliorates hypertension, heart. 2020) and SARS-CoV-2-induced lung injury in mice (failure (2021). We show

therapeutic effects of B38-CAP on abdominal sepsis- or acid aspiration-induced acute lung injury. ACE2
expression was downregulated in the lungs of mice with cecal ligation puncture (CLP)-induced sepsis or acidinduced lung injury thereby leading to upregulation of Ang II levels. Intraperitoneal injection of B38-CAP
significantly decreased Ang II levels while upregulated angiotensin 1‒7 levels. B38-CAP improved survival rateof the mice under sepsis. B38-CAP suppressed the pathologies of lung inflammation, improved lung dysfunction
and downregulated elevated cytokine mRNA levels in the mice with acute lung injury. Thus, systemic treatment
with an ACE2-like enzyme might be a potential therapeutic strategy for the patients with severe sepsis or ARDS(

2022).

Copyright © The Japanese Pharmacological Society1-B-YIA04-4

一般演題(YIA)

Molecular basis of sex difference in sepsis〇岩鶴 果奈、坂本 多穗、竹下 舜也、山下 ほのか、黒川 洵子
静岡県立大・薬・生体情報

Male sex is more prone to septic death. However, its molecular mechanism remains unknown. Our studies
revealed that inflammatory responses in skeletal muscle play a pivotal role in sepsis. Furthermore, recent
findings suggested that septic responses in skeletal muscle is a key of the sex differences in septic death. To
elucidate if the sex differences in sepsis is from gonadal and sex chromosomal differences, cecal ligation andpuncture (CLP)-induced septic symptoms in Four Core Genotypes (FCGs) mice (XX gonadal males or females,
and XY gonadal males or females) were investigated in this study. Our survival analysis showed that XX female
mice were significantly resistant among FCGs to septic death. Furthermore, our RNA-seq analysis in skeletal
muscle of septic FCGs revealed that different activity of inflammatory pathways and four inflammation related
genes (,,,) were overexpressed specifically in XX females. In vitro analysis using C2C12

myotubes revealed that estradiol-treatment but not testosterone-treatment enhance mRNA expressions of thesegenes. Our study suggests the involvement of interactive effects of gonadal and sex chromosomal differences insex differences in sepsis. Four genes were identified as candidate genes involved in sex difference of sepsis.

Copyright © The Japanese Pharmacological Society1-B-YIA05-1

一般演題(YIA)OpTER: a low-cost method for measuring the transepithelial/endothelial
electrical resistance TER of cell layers, which reduces research costs and theburden on us all〇吉川 慧、原田 佳奈、田中 茂、秀 和泉、酒井 規雄
広島大・大学院医系科学研究科・医歯薬学専攻 神経薬理学

Transepithelial/endothelial resistance (TER) measurement is a non-invasive method to assess the integrity oftight junctions in model cells such as the intestinal epithelium and the blood-brain barrier. This technique isessential for experiments on drug kinetics and tissue damage, but commercial devices have many limitations.
Additionally, the high-grade analyzers for long-term measurements are prohibitively expensive, raising barriers
to entry into this research field.
The open source-based experimental equipment has advantages such as cost reduction and high versatility.
There have been reports of TER instruments using 'open-source way' methods, but some of these havereproducibility problems.
We propose OpTER, a reproducible, and inexpensive TER measurement method. An Arduino-based measuring
circuit can be created for less than 10,000 yen. Our method enables the recording of results equivalent to thoseof a commercially available product. Along with homemade electrodes made of biocompatible metals, thisenables continuous measurement of TER in an incubator.
The circuits and program will be available, and its simple mechanism, which can be assembled by non-experts inelectrical engineering, can easily be modified to suit the researcher's objectives. This idea is a new 'OpTion' forboth amateurs and professionals.

Copyright © The Japanese Pharmacological Society1-B-YIA05-2

一般演題(YIA)

Development of a novel mouse model of immune checkpoint inhibitor-associatedmyocarditis〇新村 貴博1,2、運天 拡人2、濱野 裕章1,3、内田 和志2、友近 七海2、宮田 晃志2、合田 光寛2,4、八木 健太1,2、相澤 風花
4、石澤 有紀5、座間味 義人2,3、石澤 啓介1,2,4
徳島大学病院・総合臨床研究センター、2徳島大・院医歯薬・臨床薬理学分野、3岡山大学病院・薬剤部、4徳島大学病1

院・薬剤部、5徳島大・院医歯薬・薬理学分野

Immune Checkpoint Inhibitors (ICI) show anti-tumor activity against various types of cancer, but they also
disrupt the balance of the immune system and cause autoimmune-like adverse events. ICI-related myocarditis,in particular, has a fatality rate of over 40%, making the development of preventive and therapeutic agents anurgent priority. In present study, we developed a simple and reproducible experimental model for ICI-associatedmyocarditis.
Myocardial myosin peptide (50 µg) was administered subcutaneously to male, 8-week-old BALB/c wild-type
and PD-1KO mice at day 0 and 7. Three weeks after the initial myosin administration, the development of
myocarditis was evaluated. HE staining and Masson trichrome staining showed inflammatory cell infiltrationand ﬁbrosis in myocardial tissue in myosin peptide-treated PD-1 KO mice. Next, the involvement of CD4⁺ andCD8⁺ cells was examined by immunostaining, and the inﬁltration of CD4⁺ and CD8⁺ cells was conﬁrmed in thehearts of myosin-treated PD-1KO mice. Finally, the results of real-time PCR showed that myosin administration
tended to increase gene expression of inflammatory cytokines and fibrosis markers in the hearts of PD-1KOmice.
It is expected that this model will be used to develop new prophylactic and therapeutic agents for ICI-associatedmyocarditis.

Copyright © The Japanese Pharmacological Society1-B-YIA05-3

一般演題(YIA)

Identification of the therapeutic target for tendinopathy through the combinationof real world data analysis and pharmacological experiments〇古田 晴香1、山田 麻莉1、長島 卓也1、松田 秀一2、永安 一樹1、白川 久志1、金子 周司1
京都大・院薬・生体機能解析学、2京都大・院医・整形外科学

1Tendinopathy is a degenerative disease characterized by rupture, pain or loss of strength at tendon tissues.
Multiple intrinsic and extrinsic factors, such as aging and fluoroquinolones, are involved in the development of
tendinopathy. However, despite much work, the exact pathophysiological mechanism remains unclear. In thisstudy, we analyzed databases of self-reported adverse events and IBM MarketScan insurance claims database to
explore a coexisting drug that reduced the incidence of tendinopathy, and found that dexamethasone prevented
fluoroquinolone-induced tendinopathy. In experimental validation of the hypothesis, chronic treatment ofpefloxacin to rats caused mechanical fragility and histological changes in tendon, which were both mitigated by
the cotreatment of dexamethasone. For its molecular mechanism, in vitro studies revealed that oxidative stress
was increased in pefloxacin-treated tenocytes, which was suppressed with the cotreatment of dexamethasone.
Also, the increase in the gene expression level of glutathione peroxidase 3 (GPX3) was observed in
dexamethasone-treated tenocytes. In fact, the overexpression of GPX3 mitigated pefloxacin-induced oxidative
stress in tenocytes. These results suggest that dexamethasone reduces risk of tendinopathy by suppressing
oxidative stress through the upregulation of GPX3. This data-driven approach based on clinical evidence will
pave the way for the identification of therapeutic target for tendinopathy with high clinical predictability.

Copyright © The Japanese Pharmacological Society1-B-YIA05-4

一般演題(YIA)

Investigating molecules that promote well-being through the brain - locomotor
system interaction〇稲田 祐奈、東田 千尋
富山大・和漢研・神経機能学領域

Subjective well-being (SWB) has been shown to correlate with longevity and healthy state. Several studies
suggested that factors influencing SWB were at least physical activity, cognitive function, and social resource.
The fact that SWB, which is an emotional issue, is influenced by physical activity and cognitive activity, and viceversa SWB affects physical activity and cognitive activity, suggests that the locomotor system and cognitive
function are closely related to mental health. However, the molecular basis of these interactions has not beenclarified. We hypothesized that some molecules responsible for these interactions circulate the brain and thelocomotor system. This clinical study aimed to find molecules responsible for controlling SWB from the blood
circulation.
Subjects were healthy elderly people over 65 years old who have no functional troubles in daily life. Evaluation
items were SWB, lifestyle, cognitive function (CF), motor function (MF) and daily activity (DA). To elucidatefeatures of elder people with high SWB, subjects were divided by their SWB scores into 4 groups. High SWB
was associated with high CF, MF and DA. Comprehensive analysis of responsible molecules in plasma forcontrolling high SWB are under investigation.
There has been no molecular explanation of why physical activity, cognitive activity, and social activity affect
SWB. The present study has the potential to answer the question and to provide a new perspective for healthand longevity research with significant impact for medical, psychological, and social sciences.

Copyright © The Japanese Pharmacological Society1-B-YIA06-1

一般演題(YIA)

Development of a novel anti-EpCAM monoclonal antibody and its application forcancer diagnosis and therapy〇李 冠傑1、鈴木 裕之1、金子 美華2、加藤 幸成1,2
1

東北大・院医・分子薬理学、2東北大・院医・抗体創薬研究分野

The epithelial cell adhesion molecule (EpCAM) is a cell surface glycoprotein, which is highly expressed on
carcinoma cells. Because EpCAM is involved in cell adhesion, proliferation, survival, stemness, andtumorigenesis, it is thought to be a promising target for cancer diagnosis and therapy. Herein, we developed
anti-EpCAM monoclonal antibodies (mAbs) using the Cell-Based Immunization and Screening (CBIS) method.
One of the established anti-EpCAM mAb, EpMab-37 (mouse IgG1, kappa), reacted with EpCAM-overexpressed
Chinese hamster ovary-K1 cells (CHO/EpCAM) or a colorectal carcinoma cell line (Caco-2) in flow cytometry.
In contrast, EpMab-37 did not react with EpCAM-knocked out Caco-2 (BINDS-16) cells in both flow cytometry
and Western blot analysis. EpMab-37 could stain formalin-fixed paraffin-embedded colorectal carcinoma tissues
by immunohistochemistry. Furthermore, we converted the subclass of EpMab-37 from mouse IgG1 into IgG2a(named as EpMab-37-mG2a), and further produced a defucosylated version (EpMab-37-mG2a-f), using FUT8deficient ExpiCHO-S (BINDS-09) cells. The EpMab-37-mG2a-f administration significantly suppressed thedevelopment of Caco-2 xenograft tumors in mice compared with the control IgG. In contrast, EpMab-37-mG2a-f
did not suppress the development of BINDS-16 xenograft tumors. These results indicated that EpMab-37 isuseful for detecting EpCAM in tumors, and EpMab-37-mG2a-f could contribute to the antibody therapy for
EpCAM-positive tumors.

Copyright © The Japanese Pharmacological Society1-B-YIA06-2

一般演題（YIA）

The role of PGD2/CRTH2 signaling in allergic reaction〇木田 美聖、中村 達朗、永田 奈々恵、村田 幸久
東京大・院農学生命科学【Background & Aim】 We previously showed that prostaglandin D2 (PGD2) promotes allergic reaction byincreasing antigen-specific IgE production. In the present study, we investigated the mechanism underlying thepromotion of IgE production by PGD2 focusing on the role of its receptor, chemoattractant receptorhomologous molecule on Th2 cells (CRTH2).
【Methods & Results】 We intradermally sensitized wild type (WT) and CRTH2 deﬁcient mice () withovalbumin (OVA). The serum OVA specific IgE level and the allergic reaction against OVA stimulation werelower inthan those of WT. Immunostaining of lymph nodes showed that dendritic cells (DCs)expressed PGD2 synthase. Consistently, bone marrow derived DCs released PGD2 in response to OVA
stimulation in vitro. The OVA-sensitization increased immune cell number in lymph node and Th2 cytokine
productions from lymphocytes in WT. CRTH2 deficiency significantly decreased the immune cell number andcytokine productions. We finally revealed that intravenous transplantation of WT DCs but not T cells or B cells
restored the serum levels of OVA specific IgE production and allergic reaction in.

【Conclusion】 In summary, antigen invasion stimulates PGD2 production from DCs which promotes Th2
cytokine production in lymph node through CRTH2 signaling. These phenomena result in promoting antigen
specific IgE production.

Copyright © The Japanese Pharmacological Society1-B-YIA06-3

一般演題(YIA)

Regulation of STING signal by advanced glycation end products〇西中 崇1、ハティポール オメル ファルク1、和氣 秀徳1、渡邊 政博2、豊村 隆男2、森 秀治2、西堀 正洋3、高橋 英夫1
近畿大・医、2就実大・薬・薬理、3岡山大・院医歯薬・創薬研究推進室

1

Cyclic GMP-AMP (cGAMP) synthase (cGAS) and stimulator of interferon genes (STING) composes signal
pathway which initiates innate immunity. 2ʻ3ʻ-cyclic GMP-AMP (cGAMP) is enzymatically produced by cGAS
and activates several signaling pathway through binding to STING. Accumulating evidence indicates that cGASSTING signaling play an important role in cancer, inflammatory disease, and senescence. Advanced glycation
end products (AGEs) are biologically reactive compounds produced by prolonged exposure of proteins tocarbonyl compounds. Chemical and physiological properties of AGEs depend on type of carbonyl compound.
Accumulation of AGEs are observed in organs and tissues according to aging and leads to induction ofproinflammatory effect. Therefore, AGEs are associated with the development of inflammatory and age-relateddiseases. However, relationship between cGAS-STING signal and AGEs remains unclear. In the present study,
we investigate the effect of different types of AGEs on STING signal in macrophage. In THP-1 cells which is ahuman monocytic leukemia cell line, cGAMP transfection increased phosphorylation of TANK-binding kinase 1(TBK1) and interferon regulatory factor 3 (IRF3), resulting in upregulation of IFNβ and CXCL10.
Glycolaldehyde-derived AGEs dose-dependently suppressed cGAMP-induced the phosphorylation of TBK1 and
IRF3. In contrast, ribose-derived AGEs enhanced the phosphorylation of TBK1 and IRF3. These results may
suggest that different types of AGEs contribute to the regulation of STING signal in macrophage.

Copyright © The Japanese Pharmacological Society1-B-YIA06-4

一般演題(YIA)

Inhibitory effects of Ninjinyoeito and Juzentaihoto on myeloid-derived
suppressor cells (MDSC) stimulated by cancer cells〇近藤 雄太1、村上 一仁1、韓 立坤2、高橋 隆二2、礒濱 洋一郎11

東京理科大・薬・応用薬理、2クラシエ製薬・漢方研究所

Myeloid-derived suppressor cells (MDSC) are heterogeneous population of immature myeloid cells, that
support tumor growth, by reducing T cell activity. Therefore, drugs which can inhibit MDSC are new predictive
immunotherapeutic medicines. We have previously shown that Kampo medicines, Ninjinyoeito (NYT) andJuzentaihoto (JTT), suppress the differentiation of MDSC. In the tumor-bearing state, MDSC migrate to thetumor microenvironment (TME). In the present study, we have investigated the effects of NYT and JTT on the
migration to TME. MDSC were isolated from C57BL/6J mice and differentiated into MDSC by the treatment
with IL-6 and GM-CSF, after which the migration activity was assessed with transwell assay. The migration of
MDSC was stimulated by treatment of 4T1 cancer cells or 4T1-conditioned media, NYT and JTT significantly
inhibited the migration. Inaddition, NYT inhibits the phosphorylation of ERK1/2 in MDSC induced by 4T1conditioned media. Furthermore, NYT considerably suppressed the expression of CCR2 in MDSC. These data
indicated that, NYT and JTT suppress not only differentiation, but also the migration of MDSC to TME. This
multi-step approach in cancer treatment may be important in the immunomodulatory effects of these Kampo
medicines.

Copyright © The Japanese Pharmacological Society1-B-YIA06-5

一般演題(YIA)

Single cell multiomic analysis of PBMCs from SARS-COV-2 infected patients〇Llamas Covarrubias Mara Anais1、衣笠 泰葉1、鈴木 穣2、今井 由美子1
医薬基盤・健康・栄養研・感染病態制御ワクチンプロジェクト、2東京大・院新領域創成科学

1

Recent advances in single cell technology now allow researchers to simultaneously profile the transcriptional
program and chromatin accessibility from individual cells, providing access to both the characterization of cell
types and states, and the exploration of gene regulatory programs at the same time and in the same cells.
Here, we present an analysis of single cell transcriptomes and chromatin accessibility profiles in PBMCs from agroup of SARS-COV-2 infected subjects with a range of disease severities. We were able to identify several
immune cell types at a coarse level based on transcriptomic profiles. By additional subclustering, we found 4
clusters of CD8T cells which show differential distribution across COVID-19 severities. Analysis of DEGs
revealed higher expression of genes associated with CD8 T cell terminal differentiation and effector functions in
the cluster enriched in mild patients. Chromatin accessibility analysis of the selected DEGs in CD8T cells
confirms higher accessibility in patients with mild disease vs severe patients. Interestingly, the transcripion
factor ZEB2 was identified as one of the top markers of the mild-severity cluster. Further motif analysis will
clarify the importance of this TF in CD8T cell differentiation and outcome in SARS-COV-2 infected subjects.

Copyright © The Japanese Pharmacological Society1-B-YIA06-6

一般演題(YIA)

The development of new therapeutic method by gene analysis using feline
mammary tumor organoids〇山本 晴、臼井 達哉、佐々木 一昭
東京農工大学・農・獣医薬理学研究室【Background and Purpose】Feline mammary tumor is known to be the third most common tumors in cats andhigh grade tumors but more effective treatment has not been established. The culture method of organoids isalso familiar with a method which can maintain the epithelial tissue structures and the characteristic like stem
cell by three-dimensional culture using a special culture medium. In previous study, we created the feline
mammary tumor organoids from tissue extracted patients and revealed the suitable culture condition. Then, we
compared original tissue and organoid, and clarified the relation with the pathological characteristics or the
expressions of hormone receptor, the differences of anti-cancer drug sensitivity in each case. In this study, we
compared the gene expression of feline mammary tumor organoids with normal mammary organoids andsearched for new therapeutic targets.
【Method】RNA was extracted from organoids produced from normal mammary glands 3 cases and feline
mammary tumor 6 cases, and RNA sequencing was performed using next generation sequencer.
【Result】In RNA sequencing, there were 112 signiﬁcant diﬀerentially genes. 81 genes were markedly
upregulated in feline mammary tumor, and 31 genes were in normal mammary glands. In addition, theexpression of X genes related with estrogen receptor was significantly upregulated in feline mammary tumor. In
GESA analysis, the pathways related to apoptosis, epithelial-mesenchymal transition, and estrogen wereactivated.
【Conclusion】In feline mammary tumor and normal mammary glands, genetic diﬀerences was cleared and astrong association with estrogen was observed. We will plan to analysis the relationship between the gene
identified in this study and feline mammary tumor, and the metabolite activity by metabolome analysis using
feline serum, search for biomarkers lead to more earlier diagnosis.

Copyright © The Japanese Pharmacological Society2-B-YIA07-1

一般演題(YIA)

Development and application of a novel probe that realize the imaging analysis
of oxytocin dynamics in brain tissue〇中村 花穂
横浜国立大学大学院・環境情報学府・自然環境

Oxytocin is a peptide hormone known for its strong central actions regulating a variety of behaviors including
parenthood and social bonding to others, in addition to the long-known peripheral actions. Despite increasing
realization of its importance, dynamics and sites of action of oxytocin in the brain are poorly understood due to alack of appropriate probe; it is too small to be tagged with bulky fluorophores for visualization. Therefore, toovercome the current technical limitation and to facilitate the understandings of oxytocinʻs action in the brain,
we tried to develop and apply a new probe. To this end, we conjugated oxytocin with “alkyne-tag” via a widely
applicable simple coupling reaction. The alkyne-tag is far smaller than oxytocin, so it is expected that its tagging
will be possible without significantly changing the original properties of oxytocin molecule. After incubation
with the living brain tissues where alkyne-oxytocin behaves similarly to endogenous oxytocin, the taggedoxytocin can be specifically visualized by a click chemistry reaction. Using this probe, we conducted various
experiments to characterize the spatiotemporal dynamics of oxytocin in brain tissues. Here, I will introduce ournovel strategy and findings brought by this probe including the region-specific binding sites and dynamics ofoxytocin.

Copyright © The Japanese Pharmacological Society2-B-YIA07-3

一般演題(YIA)PDGF-BB mediates phosphate regulation in the central nervous system〇大内 一輝、高瀬 奈央子、三島 彩音、村山 佑斗、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

Idiopathic Basal Ganglia Calcification (IBGC) is a progressive neurodegenerative disease characterized bycalcification of the basal ganglia and cerebellar dentate nucleus. Several gene mutations of IBGC are reported,which are involved in phosphate transport. IBGC is thought to be caused by abnormal

includingphosphate transport. However, its mechanism is not yet clear in central nervous system. Therefore, we elucidate
the regulation of phosphate transport in neuronal cells, focusing on PDGF-BB, which is one of the causative
gene of IBGC and activates phosphate transport in vascular smooth muscle cells.
The effect of PDGF-BB on phosphate uptake was evaluated with(knockdown) SH-SY5Y cells,

which is the human neuroblastoma cell type. To determine whether the effect of PDGF-BB is mediated byorknockdown assays were conducted. To investigate the mechanism of PDGF-BB on thephosphate uptake, the expression and membrane translocation of phosphate transporters, PiT1 and PiT2
(encoded byor) was evaluated when PDGF-BB was treated.

PDGF-BB enhanced the phosphate uptake by, not. Interestingly, the activation ofphosphate uptake by PDGF-BB was not due to increase in the expression of phosphate transporters but toactivation of membrane translocation. The activation of membrane translocation by PDGF-BB was canceledwhen Akt inhibitor was treated.
PDGF-BB enhanced the phosphate uptake by the membrane translocation of PiT1, the mechanism of which isthought to be Akt signaling.

Copyright © The Japanese Pharmacological Society2-B-YIA07-5

一般演題(YIA)

GPR143, an L-DOPA receptor, in cholinergic interneurons, modulates
haloperidol-induced extrapyramidal symptoms through coupling between
GPR143 and dopamine D2 receptor〇荒井 柾美、増川 太輝、北村 慧、大瀧 百々代、五嶋 良郎
横浜市立大・院医・分子薬理神経生物学

We propose that L-DOPA by itself is a neurotransmitter. Recently, GPR143 was identified as an L-DOPA
receptor. We previously showed non-effective dose of L-DOPA potentiates behavioral response to quinpirole, adopamine D2 receptor (D2R) agonist. However, it remains undetermined whether and how GPR143 regulates
D2R-mediated behaviors. We analyzed behavioral responses to several D2R ligands using

gene-

deficient (GPR143-KO) mice. We found that haloperidol, a D2R antagonist (0.5mg/kg)-induced catalepsy wasattenuated in GPR143-KO mice compared to wild-type (WT) mice. To clarify which neural circuits that are
responsible for this phenotype, we investigated haloperidol-induced catalepsy using conditional KO mice that
expressing cre recombinase in D2R-, adenosine A2A receptor (indirect pathway)-, and in choline
acetyltransferase (cholinergic interneuron)-positive neurons. Haloperidol-induced catalepsy was attenuated in
D2R-cre (+);

and ChAT-cre;mice. Furthermore, we found that a synthetic peptide,which inhibited the interaction between GPR143 and D2R, attenuated haloperidol-induced catalepsy. These
results suggest that GPR143 expressed in the striatal cholinergic interneurons modulates haloperidol-induced
extrapyramidal symptoms through coupling GPR143 and D2R.

Copyright © The Japanese Pharmacological Society2-B-YIA08-1

一般演題(YIA)

Development of a mutant allele-specific transcriptional repressive agent in
CAG/CTG triplet repeat diseases〇松尾 和哉1、池ノ下 侑1、矢吹 悌1、川久保 厚佑1、朝光 世煌1,2、杉山 弘3、塩田 倫史1
1

熊本大・発生研・ゲノム神経、2理研BDR・脳エピトランスクリプトミクス、3京都大・院理・生物化学

Expansion of CAG and CTG (CWG) triplet repeats is causal in a number of inherited neurological diseases. The
CWG triplet repeat diseases can be broadly classified into coding and non-coding types based on their locationin the genome, such as Huntingtonʻs disease (HD) by expanded CAG repeat in a coding region and myotonic
dystrophy type-1 (DM1) by expanded CTG repeat in a 3ʻ-untranslated region. The CWG repeat diseases arethought to induce complex pathogenic mechanisms through expanded CWG repeat-derived RNAs andpolypeptides. Here we show a CWG repeat DNA targeting compound, cyclic Pyrrole-Imidazole Polyamide(CWG-cPIP) suppresses the pathogenesis of coding and non-coding CWG repeat diseases. CWG-cPIP binds tohairpin form of the mismatched CWG DNA, interfering with transcriptional elongation of RNA polymerase in a
repeat length-dependent manner. CWG-cPIP inhibits pathogenic mRNA transcripts from expanded CWG
repeat, result in reduction of CUG RNA foci and polyglutamine accumulations in DM1 and HD patient cells
and mouse models, respectively. Treatment with CWG-cPIP also ameliorates learning and memory deficits andsynaptic dysfunction in DM1 and HD mouse models with less off-target effects. Taken together, we present a
novel candidate compound that targets expanded CWG repeat DNA independent of genomic location,
demonstrating the concept of reducing the levels of pathogenic RNAs and proteins.

Copyright © The Japanese Pharmacological Society2-B-YIA08-2

一般演題(YIA)

Inhibition of heterogeneous nuclear ribonucleoprotein U suppresses astrocyte
proliferation in astroglial scar formation after spinal cord injury〇全 麗麗、村松 里衣子
国立精神・神経医療研究セ・神経研究所・神経薬理研究部

Spinal cord injury (SCI) is a devastating trauma that results in a severe disability and irreversible motor andsensory dysfunction, whereas efficient therapies have not been fully developed. After SCI, astrocytes are thepredominant cellular component that proliferates around the lesion core and contributes to the glial scarformation, which has long been considered one of the primary causes of spinal cord regeneration failure.
However, the molecular mechanisms underlying the proliferation of astrocytes in response to central nervous
system (CNS) injury remain unclear. In this study, we found that heterogeneous nuclear ribonucleoprotein U(Hnrnpu), a DNA/RNA binding protein, regulated astrocyte proliferation after SCI. siRNA-mediatedknockdown of Hnrnpu suppressed the primary astrocyte proliferation without affecting the cell viability
Moreover,.

, inhibition of Hnrnpu expression by intraspinal injection of AAV5-Hnrnpu shRNA under thecontrol of the astrocytic glial fibrillary acidic protein (GFAP) promoter inhibited astrocyte proliferation,
increased lesion size, and suppressed functional recovery of mice after SCI. Taken together, Hnrnpu exerts a
crucial role in astrocyte proliferation, where its changes would be regarded as a hallmark of CNS diseases andinjuries in which astrocytes are involved.

Copyright © The Japanese Pharmacological Society2-B-YIA08-3

一般演題(YIA)

Developmental intracerebral hemorrhage induces microglial heterogeneity〇河野 玲奈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

Microglia arise from embryonic yolk sac and colonize the brain parenchyma, followed by the maturation of
microglia-specific gene expression. Single-cell transcriptional analysis has revealed that microglia aretranscriptionally heterogeneous. However, the environmental factors that induce microglial heterogeneity arelargely unknown. Here we found that developmental intracerebral hemorrhage induces microglial heterogeneity.
In neonatal mice, we found that a portion of microglia phagocytose the red blood cells (RBCs) and
expressed, which encodes Heme Oxygenase 1, significantly higher than non-RBC-phagocytic microglia.

To examine the effect of RBC phagocytosis on the transcriptional property of microglia, we labeled Hmox1expressing microglia with red fluorescent protein by developing a transgenic mouse line, finding that microgliathat underwent RBC phagocytosis expressed genes typical for yolk sac microglia in the second postnatal week.
Thus, this study reveals that neonatal environmental factors induce microglial heterogeneity.

Copyright © The Japanese Pharmacological Society2-B-YIA08-4

一般演題(YIA)

Pharmacological analysis of behavioral addiction by quantifying the motivation
for wheel-running in mice

〇西谷 直也1,2、谷口 遥海2、小久米 泰祐2、金田 勝幸1,2
1

金沢大・院薬・薬理、2金沢大・薬・薬理In behavioral addiction, addicts repeat certain behaviors, such as internet gaming and gambling, despite
negative consequences. No cure has been established due to the lack of animal models to explore its neuronalmechanisms. Here, by focusing on voluntary wheel-running in rodents, we have developed a novel operant
conditioning task as a model of behavioral addiction, in which male C57BL/6J mice (> 7 weeks old) wereallowed for wheel-running after certain numbers of nose pokes. In this task, the magnitude of motivation forwheel-running is quantifiable by evaluating the number of nose pokes.
We first measured dopamine (DA) release using fiber photometry with GRAB-DA sensors and found that DA
was increased in the nucleus accumbens (NAc) immediately after nose-poking. Systemic administration ofantagonists for DA D1 receptor (SCH23390; 0.025‒0.1 mg/kg), D2 receptor (raclopride; 0.1‒0.6 mg/kg), or anagonist for adenosine A2A receptor (CGS21680; 0.05-0.1 mg/kg) dose-dependently decreased the number ofnose pokes. Intra-NAc infusion of these drugs (SCH23390; 0.05 µg/side, raclopride; 0.3 µg/side, CGS21680; 1
ng/side) also reduced the number of nose pokes. Additionally, systemic administration of serotonin (5-HT)2A
receptor antagonist (volinanserin; 0.01‒0.1 mg/kg) or 5-HT2C receptor antagonist (SB242084; 0.3‒1.0 mg/kg)
dose-dependently decreased the number of nose pokes. These results suggest that neurotransmission via D1,
D2, and A2A receptors in the NAc and 5-HT2A and 2C receptors are involved in the motivation for wheelrunning.

Copyright © The Japanese Pharmacological Society2-B-YIA08-5

一般演題(YIA)

The mechanisms of which microglia are related to the pathology in the "primary
astrocytic disease" Alexander disease.

〇齋藤 光象1,2、繁冨 英治1,2、篠崎 陽一1,2、パラジュリ ビジェイ1,2、小泉 修一1,2
1

山梨大・院医・薬理、2山梨大・GLIAセンター

Alexander disease (AxD) is an intractable neurodegenerative disorder caused bymutations. AxD

astrocytes show several abnormal phenotypes. Our previous study has shown that astrocytes in AxD model miceshow aberrant Ca2+ signal that was a cause of etiology of AxD. In addition, using 2 photon imaging and Iba1GCaMP6-60TM, an AxD model for microglial Ca2+ imaging, we recently found that microglial Ca2+ signals werealso dramatically enhanced in the AxD model with more frequent Ca2+ signals in both the processes and cellbodies. Such increases in Ca2+ signals were inhibited by P2Y12R antagonist but not by TTX, suggesting thatthese enhancement should be independent of neuronal activity, but dependent on extracellular ATP-mediatedsignals. Thus, we hypothesized that these microglial abnormal Ca2+ signals would be caused by increase of ATP
amount released from astrocytes. Our analysis data of scRNAseq suggested that some astrocyte subclusters
unique to the AxD model exhibit the lower expression level of the gene of astrocyte-specific ectonucleotidase
subtype. In

ATP imaging using

injection of AAV GfaABC1D ATP1.0, the signals of locally puffed

ATP persisted longer in acute slice in AxD model than control WT mice, indicating a delay of ATP degradationin AxD brain that could cause the hyperactive Ca2+ signals in microglia. To study if these P2Y12R-mediated Ca2+
signals in AxD microglia play any significant roles in the mechanism of pathology, P2Y12R antagonist wasadministered. AxD model with treatment of P2Y12R antagonist showed an exacerbation of pathological markers.
This suggested that microglia play a protective role in AxD pathology via P2Y12R. Our findings hold promise forthe future development of therapies based on microglial manipulation.

Copyright © The Japanese Pharmacological Society2-B-YIA09-1

一般演題(YIA)

Effects of fluoxetine on stress-induced neuronal activity in the ventral
hippocampus〇塩崎 裕美1、久我 奈穂子1、池谷 裕二2,3、佐々木 拓哉1,2
東北大・院薬・薬理学分野、2東京大・院薬、3国立研究開発法人 情報通信研究機構 情報通信融合研究センター

1

Fluoxetine is a selective serotonin reuptake inhibitor (SSRI) and exerts antidepressant effects, which has been
considered to be mediated by increasing serotonin concentration in the inter-synaptic cleft. However, themonoamine hypothesis has not been fully supported as there is a time lag between transient increases in brain
serotonin levels after SSRI treatment and the expression of antidepressant effects. The temporal inconsistency
may be partly reconciled by memory mechanisms, especially in the hippocampus. A theory of memory suggeststhat learned memory needs to be consolidated into neuronal circuits by repeated reactivation of memoryencoding neuronal activity. Our previous study demonstrated that the inhibition of the ventral hippocampus
after stress experiences inhibits subsequent depression-like behavior in mice. Especially, we found that sharp
wave ripples (SWRs), which represent synchronized neuronal spikes in the ventral hippocampus, are a primaryneuronal activity pattern to mediate this effect. Here, we tested whether fluoxetine affects SWRs in the ventral
hippocampus in resting and depression model mice and found that fluoxetine administration significantly
reduced ventral hippocampal SWRs in both resting mice and mice that received social defeat stress. These
results demonstrate that fluoxetine inhibits SWR-induced neuronal reactivation, suggesting an antidepressanteffect mediated by suppressing memory consolidation.

Copyright © The Japanese Pharmacological Society2-B-YIA09-2

一般演題(YIA)

Neuronal activity of claustral populations during anxiety-related behaviors isaltered by exposure to stress〇田沼 将人1、宮司 渓汰1、大久保 仁1、野村 洋2、南 雅文3、橋本 均1,4,5,6,7、笠井 淳司1
大阪大・院薬・神経薬理学分野、2名古屋市立大・院医・認知機能病態、3北海道大・院薬・薬理、4大阪大・院連合小児1

発達・子どものこころセンター、5大阪大・先導的学術研究機構、6大阪大・データビリティフロンティア機構、7大阪大・院
医・分子医薬We have recently reported that excitatory neurons in the claustrum mediate anxiety responses to acute
psychological stressors that induce negative emotional states. However, it is unclear how claustral neurons
represent information related to anxiety responses to a stressor. To address this question, here we performed
calcium imaging of GCaMP6f-expressing claustral neurons in freely moving mice during three behavioral tests;
the elevated plus maze, the open field test, and a second open field test after an exposure to a ten-minute single
social defeat stress. Prior to exposure to a stressor, we found that a subset of claustral neurons displayed anincrease in calcium levels upon transitioning to areas associated with increased anxiety in the elevated plus mazeand the open field. In the open field test after exposure to social defeat stress, a different subset of neurons,
including neurons that were activated by of stress, exhibited sustained high levels of calcium when entering andexiting the less anxiogenic corner zones of the open field. These results suggest that stress-related anxiety
information is represented in a claustral neuronal population that is different from the population representing
anxiety under non-stressed conditions.

Copyright © The Japanese Pharmacological Society2-B-YIA09-3

一般演題(YIA)

Role of ER stress-regulated high-temperature requirement A1 (HTRA1) in thefunction of placental cells in hypertensive disorder of pregnancy〇吉田 佳乃子1、草間 和哉1、安曇 麻奈1、小島 淳哉2、吉江 幹浩1、加藤 聖子3、田村 和広11

東京薬科大・薬・薬理学、2東京医科大・医・産科婦人科学、3九州大・医・産科婦人科学

Hypertensive disorder of pregnancy (HDP) affects about 10% of pregnant women, which may be caused by
dysfunction of placental trophoblasts following impaired uterine spiral arterioles. Our previous analysis revealed
that endoplasmic reticulum (ER) stress signaling may be altered in HDP, compared to normal pregnancy. In
addition, the expression of high-temperature requirement A1 (HTRA1), a serine protease was decreased in the
HDP placenta. However, a role of ER stress and HTRAs in HDP pathophysiology remains unknown. The
relationship between of ER stress and HTRAs was explored in vitro human trophoblast cells. Treatment with ER
stress inducers thapsigargin or tunicamycin increased the expression of HTRA1 and its subtype HTRA3, but did
not alter HTRA2 and HTRA4 in trophoblasts. In vitro invasion assay revealed that either thapsigargin ortunicamycin treatment and the knockdown of HTRA1 or HTRA3 inhibited trophoblast invasion. In addition, ER
stress inducers or knockdown of HTRA1 altered the ratio of soluble fms-like tyrosine kinase-1 (sFLT1) toplacental growth factor (PGF), a severity index of HDP. The expression of HTRA1 was lower in HDP placenta
compared to the normal placenta tissues. These results suggest that ER stress may regulate trophoblast invasion
partly via HTRA1 and HTRA3 and is involved in the pathogenic mechanism of HDP. It might be possible todevelop a therapeutic means that targets HTRA1 to improve pregnancy complications such as HDP.

Copyright © The Japanese Pharmacological Society2-B-YIA09-5

一般演題(YIA)

Identifying antidepressants less likely to cause hyponatremia: triangulation ofretrospective cohort, disproportionality, and pharmacodynamic studies〇長島 卓也1、早川 隆1、秋元 勇人1、皆川 貴美乃2、高橋 泰夫2、浅井 聰1,2
日本大・医・生体機能医学系薬理、2日本大・医・臨床試験研究セ1

Antidepressants are known to cause hyponatremia, but conflicting evidence exists regarding specificantidepressants. To identify antidepressants less likely to cause hyponatremia, we conducted a triangulation
study integrating retrospective cohort, disproportionality, and pharmacodynamic studies. In the retrospective
cohort study, a significant decrease in serum sodium levels was observed for selective serotonin reuptake
inhibitors (SSRIs) and serotonin-noradrenaline reuptake inhibitors (SNRIs), whereas no decrease was found fora noradrenergic and specific serotonergic antidepressant (mirtazapine). Within-class comparison revealed no
decrease in serum sodium levels for fluvoxamine among SSRIs and milnacipran among SNRIs. In thedisproportionality analysis, a significant increase in hyponatremia reports was observed for SSRIs and SNRIs,
but not for mirtazapine, fluvoxamine, and milnacipran. Finally, pharmacoepidemiological‒pharmacodynamic
analysis revealed a significant correlation between the decrease in serum sodium levels and binding affinity forserotonin transporter (SERT), suggesting that lower binding affinity of mirtazapine, fluvoxamine, andmilnacipran against SERT is responsible for the above difference. These data suggest that mirtazapine,
fluvoxamine, and milnacipran are less likely to cause hyponatremia.

Copyright © The Japanese Pharmacological Society2-B-YIA10-1

一般演題(YIA)

Bidirectional regulation of tumor progression by the endogenous µ-opioidergic
system

〇濱田 祐輔1,2、吉田 小莉1、成田 道子2,3、須田 雪明1,2、葛巻 直子1,2、森 友久1、成田 年1,2
星薬科大・薬・薬理、2国立がん研セ・研・がん患者病態生理、3東京医科大・医学総合研・分子細胞治療

1

Opioid analgesics are widely used to manage moderate to severe pain in cancer patients. The μ opioid receptor(MOR) is expressed in not only the brain and spinal cord, but also a wide range of peripheral sites, including thegastrointestinal tract and immune cells. It has been considered that the central µ-opioidergic system is closely
associated with analgesia and euphoric effects, whereas the peripheral µ-opioidergic system is responsible forside effects such as constipation and nausea. However, little is known about the role of the endogenous µopioidergic system in the control of the innate immune response. In this study, we investigated the functionalrole of central and peripheral µ-opioidergic systems in tumor progression using a pharmacological and genetics
approach. First, we found that treatment with peripheral MOR antagonists significantly decreased Lewis lungcarcinoma (LLC)-graft compared to that in control mice. On the other hand, activation of the hypothalamic µopioidergic system using the Gq-DREADD technique significantly decreased the tumor volume compared tothat in control mice. Taken together, these findings suggest that the central and peripheral µ-opioidergic
systems may play a bidirectional role in the control of tumor progression.

Copyright © The Japanese Pharmacological Society2-B-YIA10-2

一般演題(YIA)

Bidirectional pain control by spinal noradrenaline via astrocyte-neuron
interactions〇川邉 陸1、吉原 康平1、古賀 啓祐1,2、津田 誠1
1

九州大・院薬・薬理、2兵庫医科大・神経生理

Pain transmission in the spinal dorsal horn (SDH) is regulated by descending neuronal pathways from the brain,such as noradrenergic (NAergic) neurons from the locus coeruleus. While it is known that spinal NA produces
an antinociceptive effect, we have recently shown that NA has an ability to produce pain hypersensitivity via
Hes5-expressing SDH astrocytes. However, the mechanism underlying the bidirectional effect of spinal NA
remains unknown. In this study, we showed that while intrathecal injection of NA at a low dose (NAlow) induced
pain hypersensitivity via α1A-adrenergic receptors (α1A-ARs) in Hes5+ astrocytes, the hypersensitivity was notobserved by intrathecal high-dose NA (NAhigh). The effect of NAhigh was also mediated by activation of inhibitory
interneurons via α1A-ARs. We found that NAhigh also activated β1-ARs in astrocytes that suppressed theastrocytic α1A-ARs-mediated effect. However, if α1A-ARs are expressed in inhibitory interneurons, why does
NAlow produce pain hypersensitivity? We further found that activation of astrocytic α1A-ARs increased release of
adenosine, a factor that suppresses inhibitory interneurons. Therefore, our findings indicate that NA
bidirectionally modulates pain transmission via astrocyte-neuron interactions in a concentration-dependent
manner.

Copyright © The Japanese Pharmacological Society2-B-YIA10-3

一般演題(YIA)

Regulatory mechanism of fatty acid-binding protein 3 expression via
docosahexaenoic acid during pain

〇橘 男、中本 賀寿夫、徳山 尚吾
神戸学院大・薬・臨床薬学

Fatty acid-binding protein (FABP) regulates polyunsaturated fatty acids intracellular trafficking and functions as
a signal transduction via modulation of gene expression. We have demonstrated that FABP3 protein wasobserved in microglia of the median eminence (ME) of hypothalamus and this protein was increased in the MEof pain model mice. These changes were correlated with the increment of hypothalamic docosahexaenoic acid
(DHA) levels. Here, we assessed the effect of DHA on FABP3 expression using MG6 cell, a microglia cell line.
Also, we tested the effect of FABP inhibitor on the mechanical allodynia in postoperative pain model mice. MG6
cells were cultured in Dulbecco's modified eagle medium with or without 10% fetal bovine serum (FBS) as a cellstress. FABP3 was measured by qPCR. Mechanical allodynia was assessed by von Frey test. FABP3 mRNA wasexpressed on the MG6 cell. Under the condition of serum-free media, FABP3 mRNA was also significantly
increased compared to the media with 10% FBS. This increment was suppressed by DHA (300 μM). Repeatedintraventricular injection of FABP inhibitor was significantly suppressed mechanical allodynia in postoperative
pain mice. These results indicated that DHA might be involved in the regulation of microglial FABP3, and brain
FABP might work as a regulator of pain.

Copyright © The Japanese Pharmacological Society2-B-YIA10-4

一般演題(YIA)

Mirtazapine suppresses dopamine neurodegeneration by inducing
metallothionein expression via stimulation on serotonin 1A receptor of astrocyte.

〇菊岡 亮1,3、宮崎 育子1、久保田 菜月1、前田 恵実1、香川 大樹1、守山 雅晃1、佐藤 明日香1、村上 真樹1、北村 佳久
2、千堂 年昭2、座間味 義人3、浅沼 幹人1

1

岡山大・院医歯薬・脳神経機構学、2岡山大・院医歯・臨床薬剤学、3岡山大学病院・薬剤部

Parkinsonʻs disease (PD) is one of the most prevalent neurodegenerative diseases and disease-modifying
treatment is required to inhibit the disease progression. We have previously reported that 8-OH-DPAT,
serotonin (5-HT)1A full agonist, induced expression of antioxidant metallothionein (MT) in astrocyte andexhibited protective effects against 6-OHDA-induced neurodegeneration of nigrostriatal dopamine (DA)neuron. In this study, we investigated neuroprotective effect of anti-depressant mirtazapine, as an indirect 5HT1A agonist, against dopaminergic neuronal death. Mirtazapine administration to 6-OHDA-injected hemiparkinsonian mice significantly increased MT expression in striatal astrocytes and inhibited the reduction ofnigrostriatal DA neuron. These effects were cancelled by simultaneous administration of 5-HT1A antagonist. To
explore the precise neuroprotective mechanism, we examined effects of mirtazapine using primary cultured
mesencephalic neurons and striatal astrocytes from rat fetus. Neuroprotection by mirtazapine was observed onlyin neuron-astrocyte co-cultured condition. Furthermore, MT expression in astrocyte was significantly increased
when astrocytes were treated with mirtazapine-pretreated neuronal conditioned medium (Mir-NCM).
Treatment with medium from Mir-NCM-treated astrocytes (Mir-NCM-ACM) showed dopaminergic protection
against 6-OHDA. These effects were cancelled when astrocytes were treated Mir-NCM and 5-HT1A antagonist.
Moreover, MT antibody completely cancelled the neuroprotective effects of Mir-NCM-ACM. These results
suggested that mirtazapine protected DA neurons by inducing expression and secretion in/from astrocytes via
indirect stimulation on astrocytic 5-HT1A receptor.

Copyright © The Japanese Pharmacological Society2-B-YIA10-5

一般演題(YIA)

Copyright © The Japanese Pharmacological Society3-B-HS01-1

一般演題（高得点）

Histamine promotes tube formation of vascular endothelial cells

ハティポール オメル ファルク1、西中 崇1、西堀 正洋2、渡邊 政博3、豊村 隆男3、森 秀治3、和氣 秀徳1、高橋 英夫11

近畿大・医・薬理学教室、2岡山大・院医歯薬、3就実大・薬・生体情報学【Aim】
Histamine is well known as an inflammatory mediator whereas little is demonstrated about the mechanism ofhistamine-induced angiogenesis. The role of histamine in angiogenesis was first reported in 1983 in thechorioallantoic membrane (CAM) assay. Still, the histamine-induced tube formation mechanism has not been
investigated with histamine single-acting in vitro study. In the present study, we demonstrated the effects ofhistamine on the tube formation processes of human endothelial cells.
【Method】
Histamine-induced tube formation was analyzed by the matrigel assay using the human-derived vascularendothelial cell line EA.hy926. To investigate the effects of histamine H1 receptor (H1R) antagonist and protein
kinase C (PKC) inhibitors vascular endothelial cells were stained with Calcein-AM and observed under amicroscope. In addition, the expression of angiogenesis-related factors such as vascular endothelial growth
factor (VEGF) and matrix metalloproteinases (MMPs) was analyzed by the RT-PCR method.
【Result】
Histamine concentration-dependently remarkably enhanced tube formation of EA.hy926 cells in a matrigel
assay at 16 hours. In addition, histamine-induced tube formation was entirely blocked by inhibitors of H1R and
PKC. This result suggests that H1R-PKC signaling is involved in histamine-induced tube formation. We havealso shown treatment of EA.hy926 cells with 10 µM histamine resulted in a marked upregulation of VEGF
mRNA expression with a peak at 3 hours and an inhibitor of VEGF Receptor (VEGFR) -2 suppressed
histamine-induced tube formation. Additionally, histamine stimulation induced the expression of MMP-9 and
MMP-14, which play important roles in the regulation of angiogenesis, and MMP inhibitors blocked histamineinduced tube formation.
【Conclusion】
In this study, we have shown a remarkable tube formation in vitro model induced by histamine in endothelial
cells through the H1 receptor. In addition, this action was linked to the activation of PKC, VEGF, VEGFR2, and
MMPs.

Copyright © The Japanese Pharmacological Society3-B-HS01-2

一般演題（高得点）

Epithelial BLT1 Plays an Important Role in Colonic Mucosal Wound Repair〇林 周作1、Quiros Miguel2、奥 牧人1、Parkos Charles A.2、Nusrat Asma2
1

富山大・和漢研・未病、2ミシガン大・医・病理

Acute and chronic intestinal inflammation is associated with epithelial damage, resulting in mucosal wounds inthe forms of erosions and ulcers in the intestinal tract. Intestinal epithelial cells (IECs) and immune cells in thewound milieu secrete cytokines and lipid mediators to influence wound repair. Leukotriene B4 (LTB4), a lipid
chemokine, binds to its receptor BLT1 and promotes migration of immune cells to sites of active inflammation,
however a role for intestinal epithelial BLT1 during mucosal wound repair is not known. Intestinal epithelial
BLT1 expression is increased when epithelial cells are exposed to an inflammatory microenvironment. Usinghuman and murine primary colonic epithelial cells, we reveal that LTB4-BLT1 pathway promotes epithelial
migration and proliferation leading to accelerated epithelial wound repair. Furthermore,

intestinal wound

repair experiments in BLT1-deficient mice and bone marrow chimeras demonstrate an important contributionof epithelial BLT1 during colonic mucosal wound repair. Taken together, our findings show a novel pro-repair
mechanism in IECs mediated by BLT1 signaling.

Copyright © The Japanese Pharmacological Society3-B-HS01-3

一般演題（高得点）

The prediction of therapeutic targets and microRNA network in the coronavirus
pathogenesis pathway〇田邊 思帆里1、カデール サビーナ2、小野 竜一3、カブラル オラシオ4、青柳 一彦5、広瀬 明彦1、横崎 宏6、佐々木 博
己71

国立医薬品食品衛研・安セ・安全性予測評価、2ナノ医療イノベーションセンター、3国立医薬品食品衛研・安セ・毒性、4

東京大・院工、5国立がん研・基セ・臨床ゲノム解析、6神戸大・院医・病理、7国立がん研・基セ・シーズ探索

The coronavirus pathogenesis pathway is activated in coronavirus infection. To reveal the therapeutic targets ofthe coronavirus pathogenesis, public gene expression data were analyzed in Ingenuity Pathway Analysis (IPA).
Among more than 100,000 analyses and datasets, 106 analyses and 106 datasets were related to SARS
coronavirus 2. The 49 analyses were involved in SARS coronavirus 2 and human, which comprise of 27 analyses
including 9 analyses on tissue “skin” GSE156754 and 22 analyses on lung adenocarcinoma. FOS and JUN in thecoronavirus pathogenesis pathway were activated in SARS-CoV-2 infected lung adenocarcinoma. Coronavirus
pathogenesis pathway was activated in SARS-CoV-2 infected iPS cell-derived cardiomyocytes. The molecular
networks and gene expression in diffuse- and intestinal-type gastric cancer (GC) have been analyzed as well.
Coronavirus pathogenesis pathway was activated in diffuse-type GC and inactivated in intestinal-type GC.
Telmisartan, acetaminophen and arsenic trioxide were found to interact with the coronavirus pathogenesis
pathway. NFkappaB, a target of thalidomide, was activated in diffuse-type GC. The coronavirus pathogenesis
pathway had direct relationships between microRNAs including let-7, mir-10, mir-15, and mir-155. The
molecules identified have potential to be the therapeutic targets.

Copyright © The Japanese Pharmacological Society3-B-HS01-4

一般演題（高得点）

Upregulation of neuregulin-1 in the ventricle of diabetic cardiomyopathy model
mice and its functional significance〇三上 義礼、岩瀬 奎輝、冨田 太一郎、大島 大輔、赤羽 悟美
東邦大・医・生理・統合生理

Diabetic cardiomyopathy (DMCM) is myocardium disorder in diabetic patients independent of diabetic
vascular dysfunction. It is characterized by an early diastolic dysfunction and subsequent progression to systolic
dysfunction. The underlying mechanism of the DMCM development has not yet been fully understood. We
aimed to elucidate the molecular mechanism of DMCM progression. In the streptozotocin (STZ)-induced DM
model mice 4 weeks after STZ injection (STZ-4W), diastolic function was impaired without systolic
dysfunction. In the ventricles of STZ-4W mice, the mRNA and protein expression levels of neuregulin1 (NRG
-1) turned out to be significantly higher than that of control mice. Chronic insulin administration restored theblood glucose, left ventricular diastolic function, and NRG-1 expression to the control levels. NRG-1 was
localized in the epicardium, endocardium, and endothelial cells of blood vessels in the ventricle. To clarify therole of up-regulated NRG-1 in the early stage of DMCM, we examined the effects of trastuzumab (TRZ),
antibody against NRG-1 receptor ErbB2. Not only diastolic function but also systolic function was significantly
impaired in the TRZ-injected STZ-4W mice. These results suggest that a compensatory increase in NRG-1
prevents the progression to systolic dysfunction during the early stage of DMCM.

Copyright © The Japanese Pharmacological Society3-B-HS01-5

一般演題（高得点）

Effects of esaxerenone on blood pressure and sodium balance in Dahl saltsensitive hypertensive rats

〇西山 成、服部 真依、北田 研人、ラフマン アサダ
香川大・医The nonsteroidal mineralocorticoid receptor blocker, esaxerenone, is effective in reducing blood pressure (BP)in hypertensive patients. However, the mechanism responsible for anti-hypertensive effect of esaxerenone is notclear. Therefore, we investigated esaxerenone-driven sodium homeostasis and its association with changes in BP
in Dahl salt-sensitive (DSS) hypertensive rats. BP was measured by a radiotelemetry system, and sodium
homeostasis was determined by an approach of sodium intake (food intake) and excretion (urinary excretion) in
DSS rats with a low-salt diet (0.3% NaCl), high-salt diet (HSD, 8% NaCl), HSD plus 0.001% esaxerenone(w/w), and HSD plus 0.05% furosemide. HSD-fed DSS rats showed a dramatic increase in BP with a nondipper pattern, while esaxerenone treatment, but not furosemide, significantly reduced BP with a dipper
pattern. The cumulative sodium excretion in the active period was significantly elevated in esaxerenone- andfurosemide-treated rats compared with their HSD-fed counterparts. However, a significant increase in thesodium/potassium ratio was only observed in esaxerenone-treated rats. Sodium content in the skin, skinned
carcass, and total body tended to be lower in esaxerenone-treated rats than in their HSD-fed counterparts, whilethese values were unchanged in furosemide-treated rats. Consistently, sodium balance tended to be reduced inesaxerenone-treated rats during the active period. These data indicate that esaxerenone-induced reduction in
BP is associated with improvement of body sodium homeostasis in salt-dependent hypertension.

Copyright © The Japanese Pharmacological Society3-B-HS02-1

一般演題（高得点）

mPGES-1/PGE2 axis induces recovery from ischemia via SDF-1/CXCR4 axismediated accumulation of Tregs in ischemic muscle〇天野 英樹1、江島 耕ニ2、伊藤 義也1、細野 加奈子1、畑中 公1、植松 智3、審良 静男4、成宮 周5、馬嶋 正隆6
北里大・院医療、2北里大・理・免疫学、3大阪市立大・医・ゲノム免疫学、4大阪大・生体防御部門・自然免疫学分野、5京1

都大・院医・創薬医学講座、6神奈川工科大学・臨床工学科・病態治療研究室

We have reported that mPGES-1/PGE2 induced blood flow recovery from ischemia by promoting accumulationof Tregs. Expression of chemokines and the SDF-1/CXCR4 axis has been implicated in Tregs migration inangiogenesis. Therefore, we examined whether the SDF-1/CXCR4 axis plays a role in the accumulation of Tregs
in angiogenesis.
Male 6-8 week-old wild-type mice (WT), mPGES-1-deficient mice (mPges-1-/-) were used. Recovery fromischemia was estimated by laser Doppler imaging. Contribution of Tregs was estimated by immunohistichemical
study against FoxP3.The expression of Sdf-1 and Cxcr4 in ischemic muscle and accumulated Tregs were
estimated by real time RT-PCR. The function of Tregs was estimated by in vitro suppression assay.
In WT, recovery from ischemia was significantly suppressed in CXCR4 antibody treated mice compared toControl IgG. In contrast, there was no significant changes in mPges-1-/-. In ischemic muscle tissue, expressionof Sdf-1 and Cxcr4 mRNA was significantly decreased in mPges-1-/- mice compared with WT mice. In
accumulated CD4+CD25+ Tregs in ischemic muscle tissue, expression of Cxcr4 mRNA was significantly
decreased in mPges-1-/- mice compared with WT mice. Furthermore, in vitro analysis revealed that expressionof Cxcr4 mRNA was induced in Tregs from WT mice upon anti-CD3 stimulation but not in Tregs from mPges-1-/- mice. In vitro suppression assay showed there was no difference in Treg function between WT and mPges-1-/- mice.
These results suggested that SDF-1/CXCR4 axis induces ischemic recovery by the accumulation of Tregs inischemic muscle which was depended on mPGES-1/PGE2 axis.

Copyright © The Japanese Pharmacological Society3-B-HS02-2

一般演題（高得点）

Metalloprotease Nardilysin regulates sinus node automaticity throughmodulating ion channel transcription.

〇大野 美紀子1、松浦 博2、牧山 武3、糀谷 泰彦3、西 清人1、西 英一郎11

滋賀医科大・医・薬理、2滋賀医科大・医・生理、3京都大・医・循環器内科

Nardilysin (NRDC) is a metalloprotease of the M16 family, which has multiple functions such as enhancing theectodomain shedding of membrane proteins in the extracellular space and regulating transcription in the
nucleus. NRDC-deficient mice (-/-) show various phenotypes such as hypomyelination, hypothermia, andbradycardia. In the present study, we have focused on the role of NRDC in regulating heart rate and obtained
the following results: (1) The intrinsic heart rate, determined by pharmacological blocking of the autonomic
nervous system, was significantly reduced in-/-; (2) Funny (If) current and T-type calcium current weresignificantly reduced in isolated Nrdc-/- sinus node cells; (3) Messenger RNA levels of Cav3.1 and HCN1/4, ion
channels involved in sinus automaticity, were markedly decreased in-/- hearts; (4) Gene knockdown of

NRDC in primary rat cardiomyocytes reduced HCN1/4 mRNA levels; (5) Chromatin immunoprecipitation
PCR showed NRDC binding to the promoter regions of Cav3.1 and HCN1/4; (6) Reintroduction of wild-type
NRDC, but not the enzymatically inactive mutant of NRDC (E>A mutant), into NRDC-deficient cells restored
HCN1 mRNA expression; (7) NRDC-E>A mutant knock in mice showed bradycardia and significantly reduced
intrinsic heart rate, suggesting that NRDC enzyme activity is important for the control of heart rate. Together,
our results indicate that NRDC in cardiomyocyte controls heart rate through the transcriptional regulation ofion channels critical for sinus automaticity.

Copyright © The Japanese Pharmacological Society3-B-HS02-3

一般演題（高得点）

Regulation of cardiac robustness and mitochondrial quality by sulfur metabolism〇西村 明幸1、湯 肖康1、下田 翔2、加藤 百合2、西山 和宏2、西田 基宏1,2
1

生理研・心循環、2九州大・院薬・生理学

Proper mitochondrial quality control is indispensable for cardiac homeostasis and defects in mitochondrial
dynamics are implicated in the development of cardiac diseases. Our group has investigated the molecularmechanism underlying the development of maladaptive cardiac remodeling, especially myocardial senescence,and found that mitochondrial hyperfission induced by aberrant activation of Dynamin-related protein 1 (Drp1),
a mitochondrial fission-accelerating protein, is a key determinant of cardiac remodeling and fragility.
Supersulfides have been recently recognized as a key molecule to regulate redox homeostasis and are abundantly
discovered in both prokaryotes and eukaryotes. We found that Drp1 activity is negatively regulated bysupersulfide-mediated polysulfidation of Drp1 at Cys624. Ischemic stress induced by myocardial infarction
converted supersulfides into hydrogen sulfide, and reduced supersulfides promoted Drp1 hyperactivation via
depolysulfidation of Cys624, causing myocardial senescence and cardiac fragility. Exposure of cardiomyocytes toenvironmental electrophiles such as methylmercury also induced supersulfide depletion and triggered
mitochondrial hyperfission-associated myocardial senescence.

Copyright © The Japanese Pharmacological Society3-B-HS02-4

一般演題（高得点）〇工藤 藤美1、金 恵理2、眞鍋 一郎11

千葉大・院医・疾患システム医学、2千葉大・院薬・分子心血管薬理学

The number of patients with heart failure has increased due to the aging of society. We previously showed thatcardiac Ly6Clo macrophages from aged mice lost the cardioprotective function observed in those from young
mice. Notably, our single-cell RNA-sequencing (scRNA-seq) analysis demonstrated that tissue-resident
macrophage subpopulations were replaced with aged mice-specific subpopulations. In young mice, cardiac
macrophages were constitutively expressed various chemokines, although there was little or no expression ofcytokines in the steady state. Our scRNA-seq data revealed lower expression levels
of

,

,

and

and higher expression of

in aged mice-specific cardiac macrophages. These

results suggest that alteration of the expression of chemokines in cardiac macrophages may result in changes in
the proportion of each immune cells in the heart of aged mice, triggering age-associated disease induction. We
confirm the pathophysiology of macrophage-specific-deficient mice in a transverse aortic constriction(TAC) model to elucidate the regulatory mechanism of progression of tissue fibrosis by macrophage-derived
chemokines. Furthermore, we assessed the effects of CCR5 inhibitor on tissue fibrosis in TAC mice. Thus, weclarify the relationship between aging-associated heart failure and phenotypic changes in cardiac macrophages.

Copyright © The Japanese Pharmacological Society3-B-HS03-1

一般演題（高得点）

Preceding inhibition regulates hippocampal spikes sequences〇野口 朝子1、池谷 裕二1,2
1

東京大・院薬・薬品作用、2東京大・Beyond AI研究推進機構

Neuronal spike times are essential for coding information. A subset of neurons often emits a series of action
potentials and generate sequences of spikes. The spike times of neurons during a sequence are regulated at themillisecond level while preserving flexibility to generate diverse patterns in different sequences. Spike sequences
are exemplified by memory replays during sharp wave‒ripples (SWRs). Hippocampal pyramidal cells that aresequentially activated during behavior are reactivated in time-compressed manner during SWRs while animals
are immobile or asleep. This replay of sequential activity has been believed to contribute to memory
consolidation and navigational planning. However, the mechanism for such flexible modification of spike times
remains unclear. In this study, we conducted in vivo whole-cell recordings simultaneously from up to three CA1
pyramidal cells and examined the membrane potential dynamics at the single-cell level. Neurons weretransiently hyperpolarized tens of milliseconds before SWRs. The pre-SWR hyperpolarizations varied inmagnitude across SWR events and individual neurons, and larger pre-SWR hyperpolarizations induced later
spike times during SWRs. Thus, pre-SWR inhibition coordinates the sequential spike times of CA1 pyramidalcells and diversifies the repertoire of sequence patterns.

Copyright © The Japanese Pharmacological Society3-B-HS03-2

一般演題（高得点）

Single-molecule imaging of synaptic molecules within the brain tissue〇大久保 洋平1、並木 繁行2、浅沼 大祐2、櫻井 隆1、廣瀬 謙造2
1

順天堂大・医・薬理学、2東京大・院医・細胞分子薬理学

Accumulating evidence suggests that molecular dynamics at nanometer scale is crucial for synaptic functions.
Single-molecule fluorescence imaging is a super-resolution live imaging method that enables direct tracking of
movement of individual molecules. However, conventional single-molecule imaging has been applicable only todissociated cells on coverslips due to technical limitations, preventing the analysis of events that occur only in
the intact brain tissue. In this study, we set out to develop a method for single-molecule imaging within brainslices and the brain. We developed and employed a novel chemical tag technology named De-QODE.

This technology consists of a small-molecular QODE probe and DeQODE protein tag. Non-fluorescent QODE
becomes highly fluorescent upon reversible binding to DeQODE. These properties realize fluorescent labelingof proteins of interest with extremely low-background fluorescence even within tissue samples. Furthermore,
De-QODE-based labeling is repeatable after photobleaching. Owing to De-QODE-based single-molecule
imaging, we succeeded in high-density tracking of synaptic molecules in pyramidal neurons deep within acutecortical slices.

Copyright © The Japanese Pharmacological Society3-B-HS03-3

一般演題（高得点）

Mechanism of aggregation of α-Synuclein initiated by RNA phase transition〇矢吹 悌1,2、松尾 和哉1、酒井 勇輔1,2、塩田 倫史1,2
1

熊大・発生研・ゲノム神経、2熊本大・薬The mechanism underling dysfunction of cellular proteostasis on α-synuclein (α-Syn) leading to pathogenesisof synucleinopathy remains unclear. Recently, we reported that the binding of an RNA secondary structure Gquadruplex (G4RNA) to a prion-like protein FMRpolyG causes its liquid-to-solid phase transition, leading toneurodegeneration in a hereditary neurodegenerative disease, Fragile X-associated Tremor/Ataxia Syndrome(FXTAS) (2021). Here, we introduce the possibility that G4RNA is a key pathogen on the phasetransition of α-Syn. Purified α-Syn protein binds to G4 structure formed RNA specifically, and that theaddition of G4RNA promoted the liquid-to-solid phase transition on α-Syn under molecular crowding
In mouse primary neurons, G4RNA assembly was immediately observed under cellular stress conditions,
thereafter co-aggregation of α-Syn with G4RNA was occurred. Artificial assembly of G4RNA using an
optogenetic approach initiated α-Syn aggregation, thereby elicits neuronal dysfunction in mouse primaryneurons. These results suggest that G4RNA assembly evoked by various cellular stress triggers to develop
aggregation of α-Syn, which may be a cellular mechanism underlying onset of sporadic synucleinopathy. We
now analyze relationship between G4RNA and α-Syn aggregation.

Copyright © The Japanese Pharmacological Society.

3-B-HS03-4

一般演題（高得点）

Clarification of molecular mechanisms for axonal regeneration in the brains of
Alzheimer‘s disease model mouse〇楊 熙蒙、東田 千尋
富山大・和漢研・神経機能学

Alzheimerʻs disease (AD) is a neurodegenerative disorder characterized by Aβ deposition and disruption ofneural networks in the brain. We previously found that diosgenin, a constituent of Dioscorea Rhizoma, restoredAβ-induced axonal atrophy in neurons () and recovered memory deficits in a mouse model of AD,

5XFAD. Importantly, we were the first to discover that diosgenin administration promoted long-distance axonal
regeneration in 5XFAD mice brains. In the present study, we aimed to clarify molecular mechanisms forcontrolling accurate pathfinding of injured axons in AD brains.
Axon-regenerated neurons (after diosgenin administration) in the neural circuits contributing memory
formation; from the hippocampus to the prefrontal cortex, were selectively visualized by retrograde tracings.
Naïve neurons and axon-regenerated neurons in the brain slices were separately captured by laser
microdissection to serve DNA microarray. Overexpression of the gene, whose expression level was drasticallyelevated in axon-regenerated neurons, to the hippocampal neurons promoted axonal regeneration in the brain
and recovered memory deficits in 5XFAD mice.
Our study identified key molecules for promoting axonal regeneration toward long distance away target area in
AD brains. This finding proposes a novel therapeutic strategy for AD treatment.

Copyright © The Japanese Pharmacological Society3-B-HS03-5

一般演題（高得点）

Regulation of antinociceptive tolerance to morphine by RTP4, an endogenous
chaperone protein〇藤田 和歌子1、内田 仁司2,3、川西 雅史4、黒岩 祐介4、阿部 学2,5、崎村 建司2,5
1

長崎大・院医歯薬・医薬理、2新潟大、3新潟大、4長崎大・薬・創薬薬理学分野、5新潟大

Previous studies have shown that receptor transporter protein 4 (RTP4), one of the receptor chaperone
proteins, contributes to maturation and membrane trafficking of novel opioid receptor heterodimers consistingof mu (MOPr) and delta (DOPr) opioid receptors (MOPr-DOPr). Although MOPr-DOPr is shown tocontribute to the mechanism of development of antinociceptive tolerance to morphine, the role of RTP4 in suchmechanism has not been elucidated yet. Since

will be upregulated by repeated administration of morphineespecially in the hypothalamus, here we determined the effect of knockdown of RTP4 in the selective brain
region on the development of antinociceptive tolerance to morphine by using Rtp4flox/flox mice. In this study,
Rtp4flox/flox mice were generated and infected with AAV expressing Cre recombinase. Knockdown oflevels in

hypothalamus partially but significantly suppressed the development of morphine tolerance. In addition, we
found that the induction ofgene by MOPr-activation was reversed by inhibitors of Gi and MAPK pathways

in neuronal cells. These results indicate that RTP4 in hypothalamus partly but significantly contributes to themechanism of development of morphine tolerance after repeated administration of morphine. Also theupregulation of

by morphine may be mediated by MAPK pathways.

Copyright © The Japanese Pharmacological Society3-B-HS04-1

一般演題（高得点）

Electrophysiological properties of a mutation in the human Cav3.1 T-typechannel associated with neuropsychiatry.

〇酒井 勇輔1,2、矢吹 悌1,2、塩田 倫史1,2
1

熊本大・発生医学研究所・ゲノム神経学分野、2熊本大・薬

Low-threshold T-type calcium channels (approximately at -60 mV) show unique electrophysiological featureswith the fast inactivation and slow deactivation kinetics. T-type calcium channels are expressed in the
mammalian brain and involved in the pathophysiology of epilepsy, pain and sleep. Recently, a novel de novo
mutant Cav3.1 T-type calcium channel at V1330E have been reported by meta-analyses of the exome sequencesof patients with schizophrenia (Iyegbe CO et al., 2022.

), however, its electrophysiological properties arestill unknown. In this study, we aimed to compare the electrophysiological properties of mutant Cav3.1 at
between the A961T associated with cerebellar ataxia (gain-of-function) and the V1330E. Each mutant Cav3.1 at
A961T and V1330E was generated and transiently transfected in Neuro2A cells. Using the whole-cell patchclamp technique, we successfully demonstrated that mutant Cav3.1 (A961T) displays very slow inactivation
kinetics and unique changes in the steady state kinetics compared with wild-type Cav3.1 channel as well as
previously observation. We currently analyze the property of mutant Cav3.1 (V1330E) and would like to present
the data and discuss the significance of the mutation at V1330E on site.

Copyright © The Japanese Pharmacological Society3-B-HS04-2

一般演題（高得点）

Protective effect of lansoprazole against cisplatin-induced ototoxicity〇若井 恵里1、弓削 瑞葵2、池村 健治3、奥田 真弘3、西村 有平2
1

大阪大・院医・統合薬理学、2三重大・院医・統合薬理学、3大阪大学医学部附属病院・薬剤部

Cisplatin accumulates in the inner ear cochlea via organic cation transporter 2 (OCT2) and causes ototoxicity.
Lansoprazole, a proton pump inhibitor, ameliorated cisplatin-induced nephrotoxicity via inhibiting OCT2. In
the present study, we investigated the protective effect of lansoprazole against cisplatin-induced ototoxicity. In
the zebrafish study, we compared the effect of lansoprazole on cisplatin induced ototoxicity using in vivo
fluorescence imaging of the hair cells stained with YO-PRO1. The fluorescence signals in hair cells in zebrafish
treated with cisplatin dose-dependently decreased. Co-treatment with lansoprazole significantly suppressed thedecrease of fluorescence signals in zebrafish treated with cisplatin. Knockout of a zebrafish homolog of OCT2
also ameliorated the reduction of fluorescence signals in hair cells in zebrafish treated with cisplatin. We thenretrospectively analyzed the medical records of Mie University Hospital to examine the otoprotective effect of
lansoprazole. The incidence rate of ototoxicity was significantly lower in patients co-treated with LPZ comparedto those without lansoprazole. These results suggest that lansoprazole should suppress cisplatin-induced
ototoxicity by inhibition of OCT2.

Copyright © The Japanese Pharmacological Society3-B-HS04-3

一般演題（高得点）

Upregulation of Galectin-7 in specific tumor microenvironment contributes tosquamous cell carcinoma metastasis〇安 健博1、長岐 雄志2、星崎 みどり1,3、山口 智和1、久世 裕太4、今井 由美子3、本山 悟2、鈴木 穣4、久場 敬司1
1

秋田大・院医・分子機能学・代謝機能学講座、2秋田大・院医・地域がん医療学講座、3医薬基盤・健康・栄養研、4東京

大・大学院新領域創成科学研究科・メディカル情報生命専攻生命システム観測分野

Metastasis predicts poor prognosis in patients with esophageal squamous cell carcinoma, whereas the regulatory
mechanism of metastasis remains largely unknown. In this study, by using murine tumor metastasis model andspatial transcriptome analysis, we aimed to decipher the molecular basis of metastasis. In a syngeneic mousesquamous cell carcinoma (SCC) model of NR-S1M cells, we isolated metastasized NR-S1M cells from lymph
nodes in tumor-bearing mice and established metastatic NR-S1M cells in in vitro culture. RNA-seq analysis
revealed that interferon gene signature was markedly downregulated in metastatic NR-S1M cells compared withparental cells, and in vivo NR-S1M tumors heterogeneously developed focal immunosuppressive areas featured
by deficiency of anti-tumor immune cells. Spatial transcriptome analysis (Visium) for the NR-S1M tumors
revealed that various pro-metastatic genes were significantly upregulated in immunosuppressive areas whencompared to immunocompetent areas. Notably, Galectin-7 was identified as a novel metastasis-driving factor.
Galectin-7 expression was induced during tumorigenesis particularly in the microenvironment of
immunosuppression, and extracellularly released at later stage of tumors progression. Deletion of Galectin-7 in
NR-S1M cells significantly suppressed lymph node and lung metastasis without affecting primary tumor growth.
Therefore, Galectin-7 plays a crucial role in tumor metastasis of SCC as a pro-metastatic factor in the immunesuppressed tumor areas and may be a potential target of cancer immunotherapy.

Copyright © The Japanese Pharmacological Society3-B-HS04-4

一般演題（高得点）

Effectiveness of proton pump inhibitors on obsessive-compulsive disease
discovered in real-world data and the molecular mechanism〇幡鎌 輝1、浅岡 希美2、永安 一樹1、白川 久志1、金子 周司1
京都大・院薬・生体機能解析学、2京都大・院医・システム神経薬理学

1

Obsessive-compulsive disorder (OCD) is a psychiatric syndrome accompanied with inadequate repetitive
and/or habitual actions, which often appears as an adverse effect of dopamine receptor stimulants. In this study,
to find a new therapeutic target for OCD, we analyzed human real-world data including adverse events selfreports and insurance claims and found that concomitant use of proton pump inhibitors suppressed theincidence of OCD symptoms induced by the use of pramipexole or ropinirole. In an in vivo validation of the
hypothesis, OCD-like abnormalities, such as spontaneous repetitive behavior and facilitation of habit formation
were observed in mice received repetitive injections of quinpirole (QNP; a dopamine D2 receptor agonist; 1
mg/kg; 8 days). In this OCD model, a systemic vonoprazan (Vpz; a proton pump inhibitor; 100 mg/kg i.p.) oran i.c.v. injection of Vpz (3 µg) suppressed the QNP-induced OCD-like behaviors. In ex vivo cortical slices
prepared from OCD model mice, an increase in the firing rate was observed in pyramidal neurons of lateralorbitofrontal area, which was reduced in the presence of Vpz (10 µM) or by intracellular acidification from pH
7.3 to 7.0. In primary cultured cortical neurons in which Atp4a gene (encoding proton pump) was knocked
down, a decrease in pH by Vpz was mitigated in parallel with a reduction of firing rate. These results
demonstrate that regulation of acid-base balance of orbitofrontal neurons will be a novel therapeutic target for
OCD.

Copyright © The Japanese Pharmacological Society2-B-O01-1

一般演題（口頭）

TRAb-IgM induced by Epstein-Barr virus reactivation did not inhibit TSH binding
to the receptor〇長田 佳子1、林 一彦2、市原 克則1、澤野 達哉1、プリヨノ アグン1、三明 淳一郎1、今村 武史1
鳥取大・医・薬理学・薬物療法学、2鳥取大・医・病理学

1

Objective
Thyrotropin receptor antibody (TRAb) is a causative antibody of Gravesʻ disease. Epstein-Barr virus (EBV)
persists in human B cells and occasionally reactivates. During EBV reactivation, the host B cells differentiate tobe plasma cells and produce IgM-dominant antibodies. We have previously observed that TRAb-IgM disruptes
thyroid follicular epithelial cells and does not transduce thyroid hormone-producing signals. However, it is still
unclear how it works on receptor-binding of TSH. We aimed to investigate this.
Methods
TRAb-IgM were separated from sera or EBV-reactivated culture media of peripheral blood mononuclear cells
from Gravesʻ disease patients. TSH binding-inhibitory activity of TRAb-IgM was assessed by a commercial
radio-receptor assay kit.
Results & Discussion
All TRAb-IgM samples showed gamma-ray counts that were almost twice that of the 0 standard. This meantthat two molecules of 125I-TSH bound to one TRAb-IgM binding complex because TRAb-IgM kept a TSH
receptor for the separation procedure. This result indicated that TRAb-IgM bound to almost all TSH receptors
coated in the test tubes, and did not inhibit TSH binding to TSH receptors. Although thyroid-stimulating TRAb
is IgG type, TRAb-IgM may has particular role on Gravesʻ disease.
Conclusions
TRAb-IgM did not inhibit TSH binding to the TSH receptor. TRAb-IgM does not function as an antagonist.

Copyright © The Japanese Pharmacological Society2-B-O01-2

一般演題（口頭）

Effect of high testosterone levels on endothelial function in aorta and erectile
function in rats

〇片岡 智哉1,2,3、深本 絢子2、堀田 祐志2、真川 明将2、前田 康博2、日比 陽子2,3、木村 和哲2,3
千葉科学大・院薬・薬理学分野、2名古屋市立大・院薬・病院薬剤学、3名古屋市立大・院医・臨床薬剤学

1

Testosterone is an important hormone for the physical and mental health of men. However,testosterone administration has also been suggested to adversely affect the cardiovascular system. We
investigated the effects of excessive testosterone administration on vascular endothelial and erectile function inrats.
: 12-week-old rats were divided into the following groups: Sham, castrated (Cast), castrated with
subcutaneous administration of 100 mg/kg/month testosterone (Cast+T1), and castrated with subcutaneousadministration of 100 mg/kg/week testosterone (Cast+T4). To observe the changes in testosterone level after
the administration, rats were further divided into the following groups: control; T(6.25); T(25); and T(100),
wherein the rats were subcutaneously injected with 6.25, 25 or 100 mg/kg testosterone per week. Erectile andendothelial functions were measured using intracavernosal pressure (ICP) and isometric tension.
: The ICP/MAP ratio in the Cast group (0.42 ± 0.04) was significantly lower than that in the Shamgroup (0.79 ± 0.07). The ICP/MAP ratio in the Cast+T1 group (0.73 ± 0.06) was significantly higher thanthat in the Cast group (P <0.01) and that of the Cast+T4 (0.38 ± 0.01) group was unchanged (P >0.05). The
T(25) and T(100) groups exhibited significantly lower responses to ACh than the control group at four weeks(P < 0.01). Meanwhile, the ICP/MAP ratios in the T(25) group (0.44 ± 0.07) and T(100) group (0.47 ± 0.03)
were significantly lower than that in the control group (0.67 ± 0.05) at stimulation frequencies of 16 Hz (P <0.05).
Excessive testosterone may cause endothelial dysfunction in the aorta and erectile dysfunction inrats and that the blood concentration should be monitored after testosterone administration.

Copyright © The Japanese Pharmacological Society2-B-O01-3

一般演題（口頭）

Image-based obesogenic screening using young zebrafish.

〇島田 康人1,2,3、中山 寛子3,4、臧 黎清2,3、松岡 いづみ4、幡 佳苗4、塚本 恒星5、深田 一剛5、湯浅 翔平5、熊沢 益徳5、勝崎 裕隆6、西村 訓弘3,41

三重大・院医・薬理、2三重大・先端セ・バイオインフォ、3三重大・ゼブラセ、4三重大・院地域イノベ、5ロート製薬、6三重大・院生資The obesity epidemic has been drastically progressing in both children and adults worldwide.
Pharmacotherapy is considered necessary for its treatment. Because many anti-obesity drugs have been
withdrawn from the market due to their adverse effects, the development of new drugs is still needed. Zebrafish
are ideal model animals fortesting of anti-obesity compounds, and disease models of several types ofobesity have been developed.
We developed a screening system using young zebrafish, “zebrafish obesogenic test (ZOT)”, andperformed screening using the focused natural product (NP) library. We then performed anti-adipogenic testing
using the mouse 3T3-L1 preadipocytes to make comparison with ZOT outputs.
Seven and eleven NPs reduced lipid accumulation in zebrafish visceral fat tissues and mouseadipocytes, respectively. Of these, five NPs suppressed lipid accumulation in both zebrafish and 3T3-L1
adipocytes. We confirmed that these five NPs (globin-digested peptides (GD), green tea extract (GTE), red
pepper extract, nobiletin, and Moringa (MO) leaf powder) exerted anti-obesity effects in diet-induced obese
adult zebrafish, as a conventional model (Nakayama H,.,. 2020;25:5840). In addition, we

validated that GD improved visceral adiposity in high-fat fed mice through UCP1 upregulation (Zang L,.,. 2021;8:650975). Based on the ZOT techniques, we further analyzed the gene expression profiles of
the adipose tissue in GTE-fed zebrafish (Zang L,.,

compounds in MO leaf using ZOT (Mastuoka I,.,. 2021;26:2627) and tried to discover bioactive. 2022;00:1-9.).

ZOT can be a high-throughput alternative to adult zebrafish models and can be applied for in vivo
screening to discover novel therapeutics for visceral obesity and potentially also other metabolic disorders.

Copyright © The Japanese Pharmacological Society2-B-O01-4

一般演題（口頭）

Effects of molecular hydrogen on dysbiosis and intestinal inflammation in highfat diet-loaded senescence-accelerated mice

〇高橋 さやか、中川 慶一、永田 亘、小泉 明穂、石塚 俊晶
防衛医科大【Subject】It is suggested that aging and excessive intake of fat may induce dysbiosis and intestinal
inflammatory damage additively. Previously, we revealed that the treatment with molecular hydrogen suppresses
intestinal injury in high fat diet-loaded senescence-accelerated (SAMP8) mice.
【Method】SAMP8 mice were fed control diet or high fat diet (HFD) for 14 weeks, and then the each group
was fed placebo jelly (PJ) or hydrogen-rich jelly (HRJ) for 4 weeks. After the treatment, small intestinal tissues
were harvested for morphological examination. In addition, we performed TBARS assay by using homogenizedsmall intestine to analyze peroxidation damage by measuring malondialdehyde (MDA) level. Moreover, weanalyzed alterations of microbiota composition in cecal feces by 16S rRNA gene analysis of microbiota profiling.
【Result & Conclusion】The treatment with HRJ prevented the increases of CD11b expression and MDA levelin HFD-loaded SAMP8 mice. The treatment with HRJ did not affect the abundance of Proteobacteria phylum in
HFD-loaded SAMP8 mice. However, the expressions of Gracilibacter, Lactinobactor and Marvinbryantia wereincreased in the HRJ group.
These findings suggest that treatment with molecular hydrogen may affect the microbiota profiling and suppress
intestinal inflammation and peroxidation damage in HFD-loaded SAMP8 mice.

Copyright © The Japanese Pharmacological Society2-B-O01-5

一般演題（口頭）

D-Serine increases released acetylcholine levels in interstitial fluids in rat
submandibular glands〇吉川 正信1、大久保 みぎわ2、川口 充3
1

東海大・医、2川野歯科医院、3東京歯科大

Several D-amino acids have been observed in saliva, but their origin and function remain to be clarified. In thepresent study, large amounts of D-aspartate and small amounts of D-serine and D-alanine were detected in all
three major salivary glands in rat. No other D-enantiomers were detected. Protein expression of D-amino acidoxidase and D-aspartate oxidase, the enzymes responsible for the oxidative deamination of neutral anddicarboxylic D-amino acids, respectively, and that of serine racemase, the enzyme converts L-serine to D-serine,
were detected in all three major salivary glands in rat. The N-methyl-D-aspartate (NMDA) receptor subunit
proteins NR1 and NR2D, but not NR2A, NR2B, or NR2C, were detected in all the three major salivary glands.
Perfusion of D-serine with L-glutamate through rat submandibular artery increased salivary secretion duringparasympathetic nerve stimulation in a D-serine dose-dependent manner. In vivo microdialysis applied tosubmandibular glands revealed that perfusion of L-glutamate with D-serine through the microdialysis probe
increased acetylcholine contents in interstitial fluids in the glands of anesthetized rats as compared to that with
L-glutamate alone in an NMDA receptor antagonist-sensitive manner. The present study suggests that D-aminoacids play a physiological role in salivary glands.

Copyright © The Japanese Pharmacological Society2-B-O02-1

一般演題（口頭）

TGF-β plays a role in platelet-mediated lymph-blood partitioning〇椎谷 友博、劉 歆儀、杉山 彰、吉松 康裕、平島 正則
新潟大・院医歯In the vascular system, lymphatic vessels play a cooperative role with blood vessels, but they form networks
separated from each other. It has been shown that lymph-blood partitioning requires lymphatic endothelial cell(LEC)-induced platelet activation. However, it remains unclear whether platelet activation is involved inestablishing a network of lymphatic vessels unconnected to blood vessels. In this study, we show that plateletactivation blocks misconnection of lymphatic to blood vessels in peripheral tissues. Angiography detected
lymph-blood misconnection in phospholipase Cγ2-deficient mice which lack LEC-induced platelet activation.
LEC protrusion was detected inside the blood vessel lumen which can potentially activate platelets. LEC
protrusion was immediately retracted by platelet-derived factors containing TGF-β in culture. Inhibition of
TGF-β signaling induces the formation of lymph-blood misconnection in mouse embryonic dorsal skin. Ourfindings advance understanding of peripheral lymph-blood partitioning.

Copyright © The Japanese Pharmacological Society2-B-O02-2

一般演題（口頭）

Regulation of remote spatial memory formation by neurosteroids and its diurnalchange in mice

〇清水 貴美子1,2,3、前畑 佳納子2、池野 知子2、Wang Qiuyi4、高尾 敏文4、深田 吉孝2,3
東京医科歯科大・難治研・病態細胞生物、2東京大・院理・生科、3東大・院医・疾患生命工学・動物資源、4阪大・蛋白1

研・機能発現プロテオミクス

The biological clock regulates not only sleep or hormone secretion but also higher brain functions such as
memory formation in mammals. We found that remote spatial memory was most efficient when learning
occurred at the beginning of the light period (dawn) in mice, although there was no diurnal change in recent
memory. We found that two 7α-hydroxy-neurosteroids (7α-hydroxypregnenolone [7α-OH-Preg] and 7αhydroxydehydroepiandrosterone [7α-OH-DHEA]) are involved in spatial memory maintenance. The
neurosteroids are synthesized from cholesterol by a new member of P450 hydroxylase, CYP7B1.

mRNA

was detected widely in the mouse brain with high levels in the hippocampus with diurnal change. We identified
the occurrence of 7α-OH-Preg and 7α-OH-DHEA in the mouse hippocampus after Morrisʻs water maze task
at the beginning of the light period by using LC-MS/MS.

deficiency impaired remote spatial memory,with recent memory mostly unaffected. The hippocampal dendritic spine densities were reduced inmice and no more increased by the training in-KO-KO mice. Chronic intracerebroventricularadministration of 7α-OH-Preg and 7α-OH-DHEA in-KO mice improved the spine density andremote spatial memory performance. Notably, this improvement was more significant when the mixture of 7αhydroxylated steroids was administered than the single neurosteroid administration. We concluded that the 7αhydroxylated neurosteroids are required for synaptic remodeling for long-term maintenance of spatial memory
in mice (iScience 2020).

Copyright © The Japanese Pharmacological Society2-B-O02-3

一般演題（口頭）

Plasma clearance of intravenously infused human adrenomedullin in rats withrenal dysfunction〇細田 洋司1,2、中村 任3、吉原 史樹2
1

信州大・医・分子病態学、2国循・病院・腎臓高血圧内科、3大阪医科薬科大・薬・臨床薬学教育研究セ

Aim
Plasma adrenomedullin concentrations are reportedly elevated in patients with renal failure; however, theunderlying mechanism is unclear. The aim of this study is therefore to investigate the pharmacokinetic changesof ADM in two renal dysfunction rats.
Methods and results
The pharmacokinetic parameters were calculated from individual plasma ADM concentration vs time curves
during and after 1 h intravenous infusion of hADM in rats with acute renal dysfunction by mercury chloride
treatment (RD-Ag) and bilateral renal blood flow blockage (RD-BL). At the end of hADM infusion, plasma
ADM levels in RD-Ag rats were approximately three times as high as in RD-Bl and normal control rats. We
measured a statistically significant positive correlation between ADM C60 and sCr. AUC0‒60 during continuous
hADM infusion was also significantly increased for RD-Ag than for normal controls. Plasma ADM
disappearance after the end of infusion was similar among the three groups. The total systemic clearance of RDAg was significantly lower than that of normal rats. Pharmacokinetic analysis revealed that elevated plasma
ADM in RD-Ag rats may be caused by a reduced volume of distribution.
Conclusion
These results suggest that decreased plasma ADM clearance in RD-Ag is not due to impaired renal excretionbut to a decreased volume of distribution.

Copyright © The Japanese Pharmacological Society2-B-O02-4

一般演題（口頭）

Highly sensitive detection of superoxide by the application of bortezomib〇松本 みさき1、矢部 千尋2、楳村 敦詩1
京都府立医科大・院医・病態分子薬理、2京都工場保健会総合医学研・臨床薬理学研究セ1

The specific detection of superoxide is challenging because of its higher reactivity and short-lived property.
Among the various chemical probes for the detection of superoxide, L-012, a luminol-based chemiluminescentprobe, enables the specific detection of extracellular superoxide. Here, we demonstrated that coapplication of
the peptide boronic acid proteasome inhibitor, bortezomib, with L-012 significantly increased its luminescence
without affecting the background. The increase in the ratio of L-012 luminescence by bortezomib was morethan five-fold in both NADPH oxidase-expressing cells and the xanthine oxidase-dependent cell-free superoxide
generation system. The application of MLN2238, another peptide boronic acid proteasome inhibitor, alsoenhanced the luminescence of L-012. In contrast, carfilzomib, an epoxyketone proteasome inhibitor, did notincrease luminescence, suggesting that the effects of bortezomib depend on the chemical structure of thepeptide boronic acid, but not on its pharmacological effects. The highly sensitive detection of superoxide by theapplication of bortezomib may become useful in the experimental assessment of oxidative stress and future
diagnostic applications, particularly in limited amounts of samples.

Copyright © The Japanese Pharmacological Society2-B-O02-5

一般演題（口頭）
2+

Cholinergic suppression of Ca signaling in pancreatic β-cells.

〇太向 勇、金丸 和典、飯野 正光
日本大・医Intracellular Ca2+ signal plays an essential role in insulin secretion from pancreatic β-cells. Recent reportssuggest that Ca2+ release from the endoplasmic reticulum (ER) through cholinergic receptor stimulation
mediated by parasympathetic nerves contributes to Ca2+ signals in β-cells. However, how the Ca2+ release fromthe ER shapes the intracellular Ca2+ signal remains elusive due to limitations in the methods for direct
visualization analysis. We recently developed transgenic mouse lines expressing a genetically encoded cytosolic
Ca2+ indicator, YC-Nano50, or an ER Ca2+ indicator, CEPIA specifically in β-cells. We successfully observed
periodic oscillations of both cytosolic and ER Ca2+ signals evoked by high glucose in isolated pancreatic islets.
We also confirmed a cholinergic agonist-induced decrease in ER Ca2+, i.e. Ca2+ release from the ER. Surprisingly,
during a high glucose condition, short-term cholinergic agonist application induced a transient suppression ofcytosolic Ca2+ level to the extent comparable with the resting level, despite the release of Ca2+ from the ER. Our
results suggest that parasympathetic nerves mediate suppressive regulation of Ca2+ signaling. Further analysis isrequired to reveal the physiological roles and underlying mechanisms of this unexpected Ca2+ suppression.

Copyright © The Japanese Pharmacological Society2-B-O03-1

一般演題（口頭）

Cardiac glycosides inhibit GLUT1-mediated glucose uptake and glycolysis in++
human cancer cells by targeting intracellular Na ,K -ATPase〇藤井 拓人1、加藤 瑞希1、清水 貴浩1、田渕 圭章2、清水 康晴3、竹島 浩4、酒井 秀紀11

富山大・薬・薬物生理学、2富山大・生命科学・ゲノム機能解析学、3救心製薬株式会社、4京都大・薬・生体分子認識学

Cardiac glycosides (CGs), potent inhibitors of Na+,K+-ATPase, have been used to treat congestive heart failure.
Recently, the effectiveness of CGs for cancer therapy has been suggested. Here, we examined the effects of CGs
on glucose metabolism in human cancer cells. Low concentrations (nM levels) of CGs (ouabain, oleandrin, anddigoxin) significantly decreased the expression level of glucose transporter GLUT1 in the plasma membrane of
the cancer cells. Ouabain (20-2000 nM) inhibited 2-deoxy-D-glucose uptake and lactate secretion of cancer
cells. In intracellular vesicles of human cancer cells, Na+,K+-ATPase α3-isoform (α3NaK) is abnormally
expressed. Interestingly, the knockdown of α3NaK significantly inhibited the ouabain-decreased GLUT1
expression, while the α1NaK knockdown did not. Ouabain (200 nM) inhibited the enzyme activity of α3NaK
but not α1NaK. The ouabain-induced GLUT1 decrease was significantly inhibited by a Ca2+ chelator, a Ca2+ATPase inhibitor, an NAADP antagonist, a dynamin inhibitor, and PI3K inhibitors. These results suggest that
CGs act on intracellular α3NaK and induce the NAADP-mediated Ca2+ mobilization and PI3K activation
followed by dynamin-dependent GLUT1 endocytosis. This mechanism may explain why CGs inhibit glucose
uptake and glycolysis in human cancer cells.

Copyright © The Japanese Pharmacological Society2-B-O03-2

一般演題（口頭）
2+

Cyclic AMP/PKA potentiates Ca -dependent plasma membrane translocation ofaquaporin 5〇服部 友万成、村上 一仁、礒濱 洋一郎
東京理科大・薬・応用薬理Aquaporin-5 (AQP5) is selectively expressed in the apical membrane of exocrine glands, such as salivary andlacrimal, and plays important roles to maintain their secretory functions. Because AQP5 is not regulated bystructural gating, translocation between plasma membrane and intracellular space is important for its waterpermeable function. We and other groups have been shown that intracellular Ca2+-dependent signaling increases
AQP5 translocation to plasma membrane. On the other hand, the role of cAMP/PKA-dependent signaling on
AQP5 translocation is still unclear, although phosphorylation of AQP5 by PKA has been suggested. In several
secretory cells, interaction between cAMP and Ca2+ signals regulate their function. In this study, therefore, we
examined the combined effect of Ca2+ ionophore and PKA activator on subcellular localization of AQP5. In MLE-12 cells, ionomycin alone increased AQP5 translocation to plasma membrane, whereas forskolin alone did not.
The combined effect of ionomycin and forskolin was considerably greater than that of ionomycin alone. Thisenhancement of the effect of ionomycin by forskolin was inhibited by H-89, a PKA inhibitor. Now we areinvestigating whether PKA-dependent phosphorylation of AQP5 is involved in this potentiated translocation.

Copyright © The Japanese Pharmacological Society2-B-O03-3+

一般演題（口頭）

2+

Na /Ca exchanger type 1 functions as a “brake” of hyperactivation in hamster
sperm.

〇竹井 元1、小椋 悠平2、氏原 嘉洋2、外山 史3、林 啓太郎1、藤田 朋恵11

獨協医科大・医・薬理学講座、2名古屋工業大学・大学院工学研究科、3宇都宮大学・工Mammalian sperm including human have to undergo several physiological and biochemical changes, collectively
called capacitation, to be fertilization-competent. Capacitated sperm show specialized vigorous motility called
hyperactivation. Hyperactivation is necessary to progress in the viscous oviductal environment.
We found that the fluid of oviduct, where fertilization occurs, contains higher concentration of Na+ than in the
media used for

fertilization (IVF). Therefore, we first investigated if this difference in the concentration+of Na affects hyperactivation or not. We found that increase in the Na+ concentration delays the hyperactivation
by lowering intracellular Ca2+ levels ([Ca2+]i). The Na+/Ca2+ exchanger (NCX)-specific inhibitor SEA0400
increased [Ca2+]i, and canceled the delay of hyperactivation by Na+. These results suggest that NCX is involvedin the regulation of hyperactivation. Next, we searched for the NCX isoforms expressed in hamster sperm, andfound that NCX1 mRNA and protein are expressed in the hamster testis and sperm. Lastly, we tried to detect
NCX1 activity by measuring Na+-dependent Ca2+ influx by Fura2. The Na+-dependent Ca2+ influx was detectedin hamster sperm, and was inhibited by SEA0400 at NCX1 specific concentration. Moreover, NCX1 activity was
declined in the capacitated sperm. These results showed that NCX1 functions as a “brake” of hyperactivation in
the hamster sperm, and its downregulation triggers hyperactivation. An inhibitor of NCX1 is a possible
candidate drug to facilitate IVF.

Copyright © The Japanese Pharmacological Society2-B-O03-4

一般演題（口頭）

Identification of Ryanodine receptor 1-selective agonists〇金谷 啓之1、桑島 謙1、大出 晃士1、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・合成生物学

Ryanodine receptors (RyRs) are large-conductance Ca2+ channels located at the membrane of endoplasmic
reticulum (ER) or the muscle equivalent, sarcoplasmic reticulum (SR). Three isoforms of mammalian RyRs are
expressed across tissues and regulate diverse cellular physiology. Although various chemicals have been
identified as agonists and antagonists of RyRs, the isoform-selective pharmacological manipulation is stillchallenging. In this study, we screened selective agonists of type 1 Ryanodine receptor (RyR1) and investigated
the structure-activity relationships. The ortho/meta/para-orientation and the length of alkyl chains affect theirpotency. Furthermore, experiments with RyR1 mutants revealed the amino acid residues responsible for thechannel activation by agonists. The novel agonists identified in this study are prospective tools for the isoformselective pharmacology of RyRs.

Copyright © The Japanese Pharmacological Society2-B-O03-5

一般演題（口頭）

Clarification of pharmacological features of water-soluble components isolated
from Qing-dai (Sei-tai), as a nuclear receptor AhR activator〇坪井 妙惠、村上 一仁、礒濱 洋一郎
東京理科大・薬・応用薬理

Aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor, which can suppress the
inflammatory response. Recently, several studies indicated that activating AhR can inhibits inflammatory
processes in the gut, and that AhR ligand may be a predicted drug to treat inflammatory bowel disease (IBD). In
our previous study,, a herbal medicine, considerably inhibited LPS-induced cytokine expression

in cultured gastric epithelial cells and in intestine tissue of dextran sodium sulfate (DSS)-induced ulcerative
colitis mice. Indigo, a major component of this medicine, is known to activate AhR. However, after thefractionation of, potent inhibitory effect on cytokine expression was obtained from watersoluble fraction, which doesnʻt contain indigo. This new active component ofinhibited cytokine

expression both in vitro and in vivo, as well as indigo, whereas it did not activate AhR with the same strength.
Now, we are investigating the characteristics of the pharmacological effect of this new AhR ligand isolated fromthe water-soluble fraction. It might be a new AhR ligand, which can stimulate transrepressional activity of thisreceptor.

Copyright © The Japanese Pharmacological Society2-B-O04-1

一般演題（口頭）

The involvements of sulfatide-selectin signaling in the spinal cord on
inflammatory pain

〇渡辺 俊1、森田 元樹1,2、野村 夏実1,2、尾山 実砂1,2、岩井 孝志1,2、田辺 光男1,2
北里大・薬、2北里大・薬・附属医薬研究施設

1

The altered levels of sulfatide are observed in various kinds of diseases in the nervous system. In our previous
reports, sulfatide synthase gene expression in the spinal cord was increased one day after intraplantar injectionof complete Freundʻs adjuvant (CFA). On the other hand, intrathecal injection of sulfatide led to mechanicalallodynia. Sulfatide is not only the major glycosphingolipid in the myelin sheath but also the potent selectin
activator. Thus, we investigated the effect of a selectin inhibitor, bimosiamose, on the sulfatide-induced
mechanical allodynia.
Intrathecal sulfatide injection produced an increase in mRNA expression of inflammatory cytokines such as
TNF-α and interleukin-1β in the spinal cord. Bimosiamose inhibited the increase of these cytokine gene
expressions and the mechanical allodynia produced by sulfatide, suggesting that sulfatide induced inflammatory
cytokine expression via selectin activation. Furthermore, bimosiamose also attenuated the CFA-induced
mechanical allodynia and lipid analysis using thin-layer chromatography revealed the sulfatide accumulation in
the spinal cord during inflammatory pain.
These results indicated that the accumulated sulfatide in the spinal cord enhanced selectin activation during
inflammatory pain, which resulted in mechanical allodynia.

Copyright © The Japanese Pharmacological Society2-B-O04-2

一般演題（口頭）

Orofacial neuropathic pain is elicited by structural changes in NAergic fibers〇林 良憲、篠田 雅路
日本大・歯

Microglia are known to be involved in the structural changes of synapses and axons. Microglia in the spinal cord
are critical for the development of neuropathic pain. However, it remains unclear whether microglia involve
structural changes of neurons during neuropathic pain. Here, we investigated the above possibilities using an
orofacial neuropathic pain model. After infraorbital nerve injury, the density of DbH immunofluorescence in thetrigeminal spinal subnucleus caudalis (Vc) was decreased, while DbH positive particles increased within
microglia. DbH-positive particles in microglia were positive for MHC-I. Surprisingly, MHC-I
immunofluorescence was also observed in microglial processes. To clarify whether MHC-I secretion frommicroglia induces the uptake of DbH-positive axons, exosomes were isolated from primary cultured microglia.
Intracisternal administration of exosomes from IFNgannma-stimulated microglia elicited mechanical allodynia
in the whisker pad and downregulation of DbH expression in the Vc. In contrast, exosomes from IFNgannmastimulated MHC-I knockdown microglia unchanged pain sensitivity and DbH expression in the Vc. These
results suggest that activated microglia-derived MHC-I causes the reduction of NAergic axons, culminating inenhanced neuronal activity in the Vc.

Copyright © The Japanese Pharmacological Society2-B-O04-3

一般演題（口頭）

Androgens determine sex differences of spinal microglia〇木口 倫一1、雑賀 史浩1、波多野 裕2、日野 信次朗3、鈴木 堅太郎2
和歌山県医大・薬・生体機能解析、2山梨大・生命環境・器官形成ダイナミクス、3熊本大・発生医研・細胞医

1

Several lines of evidence indicate that spinal microglia exacerbate abnormal pain processing. Recent findings
demonstrated there are significant functional differences of microglia in pain hypersensitivity between male andfemale animals, but underlying mechanisms are poorly understood. Here, we investigated whether androgens
affect sex differences of microglia in neuropathic pain model mice. Peripheral nerve injury-induced mechanicalallodynia was suppressed by the treatment of PLX3397, a microglial inhibitor, in male but not in female mice,and the effects of PLX3397 in the spinal dorsal horn of male mice was significantly greater than that of female
mice. Gonadectomy (GDX) decreased in serum testosterone concentration and mechanical pain threshold in
male mice. Susceptibility of spinal microglia for PLX3397 in GDX-treated male mice was similar to that ofnormal female mice. Moreover, intrathecal administration of colony-stimulating factor 1 (CSF1) elicitedmechanical allodynia in male mice, but not GDX-treated male mice or normal female mice. Collectively,
functional roles of spinal microglia contributing pain hypersensitivity are different between male and female,and sex-dependent characters of spinal microglia might be determined through androgen actions.

Copyright © The Japanese Pharmacological Society2-B-O04-4

一般演題（口頭）

Involvement of platelet-derived HMGB1 in oxaliplatin-induced peripheral
neuropathy (OIPN): OIPN prevention by antiplatelet agents岸本 彩野1、堂本 莉紗1、松永 浩明1、松本 亜紗菜1、坪田 真帆1、関口 富美子1、王 登莉2、西堀 正洋3、〇川畑 篤史1
近畿大・薬、2岡山大・院医歯薬・薬理、3岡山大・院医歯薬・創薬研究推進

1

HMGB1, a nuclear protein, once released extracellularly, promotes inflammation and pain. We have shown the
involvement of macrophage-derived HMGB1 in chemotherapy-induced peripheral neuropathy (CIPN) causedby paclitaxel, bortezomib or vincristine, and of non-macrophage cell-derived HMGB1 in CIPN caused byoxaliplatin (OIPN). Given the involvement of platelet-derived HMGB1 in thrombosis, we asked whether thepathogenesis of OIPN would involve platelet-derived HMGB1 in rodents. In rat platelet-rich plasma (PRP),
thrombin induced HMGB1 release from platelets. An anti-CD42b platelet-depleting antibody dramaticallydecreased platelet count and increased plasma HMGB1 levels in mice, leading to OIPN development after
subsequent oxaliplatin treatment at a subeffective dose. Splenectomy almost doubled platelet count andaccelerated OIPN development. Repeated oral administration of different antiplatelet agents, aspirin,
clopidogrel, cilostazol and ozagrel, prevented OIPN development in mice and/or rats. In rats subjected torepeated treatment with oxaliplatin, HMGB1 levels dramatically decreased in PRP, but tended to increase inplatelet-poor plasma. Together, our study provides novel evidence for a critical role of platelet-derived HMGB1
in OIPN development, and suggests the usefulness of antiplatelet agents to prevent severe OIPN.

Copyright © The Japanese Pharmacological Society2-B-O04-5

一般演題（口頭）
2+

Mirogabalin and pregabalin, α2δ subunit ligands of voltage-gated Ca channels,
suppress acute and chronic itch

〇尾山 実砂、星山 歩海、三宅 綾乃、別宮 洸亮、山田 望美、渡辺 俊、岩井 孝志、田辺 光男
北里大・薬Antihistamines and steroids, targeting peripheral tissues, have been used as first-line drugs in the treatment ofchronic pruritus. Here, we focused on mirogabalin (MGB) and pregabalin (PGB), gabapentinoids that are
widely used to treat neuropathic pain, to explore their potential antipruritic effects in several acute and chronic
itch models. MGB and PGB, when injected i.p., i.c.v., or i.t., inhibited scratching behavior induced bychloroquine (CQ; histamine-independent pruritogen) in the neck model of acute itch. MGB (i.p., i.c.v., or i.t.)
also reduced biting and licking behaviors reflecting acute itch and pain sensation, respectively, after CQ
injection into the front of the left calf, while PGB produced apparent antipruritic effects only when it was
injected i.t. in this calf model. The antipruritic effects of MGB and PGB were also examined under chronic itch
conditions induced by applying 1-fluoro-2,4-dinitrobenzene (DNFB) repeatedly to the nape of the neck. I.p.
injection of MGB and PGB, when used therapeutically or prophylactically, suppressed spontaneous scratching
behavior and the development of chronic itch, respectively. Therefore, gabapentinoids are considered to beeffective as the therapeutic agents acting on the central nervous system including the spinal cord forantihistamine-refractory chronic itch.

Copyright © The Japanese Pharmacological Society2-B-O04-6

一般演題（口頭）

Possibilities of statins in oxaliplatin-induced chronic peripheral neuropathy〇相澤 風花1、岡林 亜美2、森山 大嗣2、薗田 悠平1,2、高橋 志門1、合田 光寛1,2、八木 健太3、新村 貴博3、座間味 義人
4、石澤 有紀5、石澤 啓介1,2,3
徳島大病・薬剤部、2徳島大・院医歯薬・臨床薬理学、3徳島大病・総合臨床研究セ、4岡山大病・薬剤部、5徳島大・院医

1

歯薬・薬理学

Many patients treated with oxaliplatin experience sensory deficits in the distal extremities. This impairment may
persist for several months after treatment is discontinued, resulting in a negative impact on quality of life.
Statins, which are used to treat dyslipidemia, have a pleiotropic action, such as anti-inflammatory effect. Here
we show that statins can be a therapeutic agent for oxaliplatin-induced peripheral neuropathy via the regulationof glutathione S-transferase (GST). The mechanical hypersensitivity induced by oxaliplatin was ameliorated on
day 7 in mice repeated orally administered statins and lasted for 21 days. Furthermore, mechanicalhypersensitivity was suppressed even when statins were administered after day 7 of oxaliplatin exposure. On theother hand, statins were not effective against cold hyperalgesia. Uptake of oxaliplatin in the DRG was notinhibited by statins. Analysis of gene association databases revealed that the expression of GST family members
is regulated by statins. Co-administration of the GST inhibitor, ethacrynic acid, reversed the statin-induced
suppression of oxaliplatin-induced mechanical allodynia. Statins might be potential therapeutic agents for thetreatment of anticancer drug-induced chronic peripheral neuropathy that do not suppress the effects ofoxaliplatin.

Copyright © The Japanese Pharmacological Society2-B-O05-1

一般演題（口頭）

Visualization of water dynamics in brain tissue using multiphoton multimodal
imaging〇篠塚 崇徳1、宮澤 剛史1、唐澤 啓子1、小関 泰之2、安井 正人1、塗谷 睦生11

慶應義塾大・医・薬理、2東京大・工・電気系工学

The flow of cerebrospinal fluid is considered to be a critical factor in the clearance of wastes in the brain. The
dynamics of fluid has been investigated by several experimental methods such as fluorescence microscopy andmagnetic resonance imaging. However, because probes used for these imagings are much larger compared towater itself, the dynamics of the fluid have been poorly understood. Here, we applied a multimodal multiphoton
imaging system to the living brain tissue. Combining stimulated Raman scattering and two-photon fluorescentimaging, the system enables us to visualize spatiotemporal dynamics of deuterated water and fluorescent dyes
simultaneously at a cellular level. We demonstrate that deuterated water diffuses faster than fluorescent dyes inthe brain tissue. Detailed analysis reveals deuterated water rapidly exchanges inside and outside of cells,
whereas fluorescent dyes only diffuse through extracellular spaces. Furthermore, we find that the dynamics ofdeuterated water is robust to changes under physiological and pathophysiological conditions; there is littlechange in the spatiotemporal dynamics of deuterated water during development and ischemia whereas
fluorescent dyes are severely affected. Thus, our new approach reveals unique properties of the dynamics of thefluid in the living brain tissue.

Copyright © The Japanese Pharmacological Society2-B-O05-2

一般演題（口頭）Electrical activity imaging and drug response with super spatiotemporal
resolution in in vitro, organoid and ex vivo neural networks〇鈴木 郁郎東北工業大Electrical activities of neuronal network with high-spatio-temporal resolution is useful for understanding brain
functions and elucidating neurological disorders, and for drug screening and safety assessment of drugs.
Complementary metal-oxide semiconductor micro-electrode array (CMOS-MEA) is excellent in detecting
detailed electrical activity patterns of neural networks due to the large number of electrodes. In this study, weinvestigated the electrical activity characteristics of brain slices, brain organoids, and cultured neural networks
using CMOS-MEA with 236,880 electrodes, which have the highest specifications in the world. In themeasurement of brain slices, we succeeded in measuring the interregional propagation of the hippocampus andcerebral cortex area in detail, and detected changes of propagation patterns due to drug administration. In
sensory neuron measurement, calculation of axon conduction velocity in single neuron and drug responses
based on firing pattern of each neuron were detected. In human iPS cell-derived central nervous system
networks and human cerebral organoids, network activity was detected on a cell-by-cell basis, and changes inpropagation patterns due to drug administration were detected. It was found that CMOS-MEA with 236,880
electrodes and a large measurement area can measure the electrical activity characteristics of ex vivo and in vitro
neural networks and single neuron in detail. It was suggested that big data with high temporal resolution iseffective for elucidation of neural circuit function and drug evaluation based on new neural activity information.

Copyright © The Japanese Pharmacological Society2-B-O05-3

一般演題（口頭）

The potentiation of photic response in the suprachiasmatic nucleus bylactoferrin

高津 麻衣、佐藤 可那江、〇守屋 孝洋
奥羽大・薬Lactoferrin (LF) is reported to have various bioactivity such as bone growth stimulation and analgesic activity.
Recently, LF has been reported to recover the fatigue in humans after the shift-work. In this study, we examinedthe effect of administration of LF on the functions of the circadian clock such as the free-running rhythm as wellas the light pulse-induced phase shift of the behavioral rhythm using mouse model. Since an orally-administered
LF is known to be largely degraded in stomach, we used the enteric microcapsule bovine lactoferrin (eMC-LF)
which protects from stomach digestion, but can be absorbed in the small intestine. The wheel-running activityof mice at 6 min-bin was automatically recorded in PC and was analyzed by CLOCK Lab. The level of mRNA ofseveral clock-related genes was quantified by in situ hybridization method. We found that oral single
administration of eMC-LF at ZT24 promoted the re-entrainment of wheel-running rhythm to 8 hr advanced LD
cycle in mice. Both acute and chronic administration of eMC-LF potentiated the light pulse-induced phase shiftof the wheel-running rhythm. Furthermore, eMC-LF increased the light pulse-induced expression ofmRNA in the restricted area of the suprachiasmatic nucleus (SCN). We also found that eMC-LF potentiated themRNA upregulation by i.c.v. administration of gastrin-releasing peptide (GRP), a neuropeptide involvedin the photic signaling in the SCN. These results suggest that LF promotes the light entrainment of the mousecircadian clock.

Copyright © The Japanese Pharmacological Society2-B-O05-4

一般演題（口頭）

Engulfment of Amyloid β-protein in neurons and astrocytes mediated by MEGF10〇藤田 融1、前田 智司2、駒野 宏人3、田邉 由幸1
1

横浜薬科大・薬・薬理学研究室、2日本薬科大・薬・生命医療薬学分野、3北海道大・院医薬・認知症先進予防・解析分

野

Amyloid-β proteins (A β), including Aβ42 and A β 43, are known pathogenesis factors of Alzheimer's
disease (AD). Unwanted substances in the brain, including A β, are generally removed by microglia, astrocytes,
or neurons via a phagocytosis receptor. We observed that neurons and astrocytes engulfed A β 42 and A β 43,
which are more neurotoxic than A β 40. We previously showed that multiple-EGF like domains 10 (MEGF10)
that is the mammalian homologue of Draper, a phagocytosis receptor of apoptotic cells in, and is thetype I transmembrane protein plays an important role in apoptotic cell elimination and is expressed in
mammalian neurons and astrocytes. Therefore, we assessed whether MEGF10 is involved in A β42 and A β43
engulfment in MEGF10-expressing neurons and astrocytes. We found that MEGF10-expressing astrocytes andneurons engulfed A β42 and A β43 but not A β40. Furthermore, incubation of the neurons and astrocyteswith A β42 and A β43 augmented MEGF10 phosphorylation; however, incubation with A β40 did not havethis augmenting effect. Our findings suggest that MEGF10 plays a phagocytosis receptor function for A β42
and A β43 in neurons and astrocytes.

Copyright © The Japanese Pharmacological Society2-B-O05-5

一般演題（口頭）

Evaluation of acute toxicity of oxaliplatin and therapeutic candidates byextracellular potential measurement using rat primary dorsal root ganglion cells〇有馬 太陽1,2、寺薗 英之1,2、栗原 崇3、小八重 薫子2、武田 泰生1,2
1

鹿児島大学病院・薬剤部、2鹿児島大・院医歯薬・薬物動態制御学、3鹿児島大・院医歯薬・生体情報薬理学

It is known that peripheral neuropathy as an unwanted side effect of the anticancer drug oxaliplatin, especiallycold allodynia, which reacts hypersensitively to cold stimuli, greatly impairs the QOL of patients. Therefore, it isdesired to develop preventive or therapeutic medicines for cold allodynia. Previous studies have shown that
pituitary adenylate cyclase-activating polypeptide (PACAP) antagonists have potent effects in preventing cold
allodynia in in vivo studies. In this study, we evaluated the adverse effects of oxaliplatin and their protective
effects using an extracellular potential measurement system using rat E14 primary dorsal root ganglion cells(DRG). The effect of oxaliplatin and antagonist of PAC1, that is a PACAP receptor, on extracellular action
potential was verified. The firing frequency of extracellular action potential increased in a concentrationdependent manner with oxaliplatin. On the other hand, the addition of PAC1 antagonists abolished the firing of
the extracellular potential. Moreover, the firing of PAC1-treated DRG neuron decreased compared to before
addition of oxaliplatin. The evaluation system for peripheral neuropathy caused by anticancer drugs bymeasuring the extracellular potential of DRG was able to show the usefulness as a system that can evaluate side
effects in the acute phase.

Copyright © The Japanese Pharmacological Society3-B-O06-1

一般演題（口頭）

Effects of Monosulfide and Trisulfide on Pathological Events in Mouse Model ofIntracerebral Hemorrhage

Atef Yara1、木下 慶大1、市原 侑晟1、牛田 啓介2、倉内 祐樹1、関 貴弘1、香月 博志1
1

熊本大・院生命・薬物活性、2熊本大・薬・薬物活性Intracranial hemorrhage (ICH) is a type of hemorrhagic strokes that is caused due to bleeding in the brain
parenchyma. Here we investigated the effects of Na2S and Na2S3 on ICH in mice. Na2S and Na2S3 were injected
into two separate groups of mice (25 µmol/kg i.p.) 30 min before induction of ICH by collagenase injection into
the striatum. Both agents attenuated the neutrophil infiltration (MPO), inhibited the upregulation of
monocyte/macrophage chemokine (CCL-2) expression and maintained the axonal fibers transport function(APP). Moreover, pretreatment with Na2S improved the motor function, prevented the decrease in the neuronalcount (NeuN), maintained the integrity of axonal fiber structures (neurofilament-H) and lowered theupregulation of neutrophil chemokine (CXCL-2) expression after induction of ICH. On the other hand,
pretreatment with Na2S3 significantly attenuated the activation of microglia/macrophages (Iba-1) in the
perihematomal area. Both agents failed to prevent ICH-induced increase in the brain vascular permeability or tolower the pro-inflammatory cytokine (IL-6) expression significantly. In conclusion, these results suggest thatNa2S exhibits more potent neuroprotective effect than Na2S3 and promotes recovery of neurological functions
after ICH.

Copyright © The Japanese Pharmacological Society3-B-O06-2

一般演題（口頭）

Involvement of Protein tyrosine phosphatase delta (PTPd) in the corticalpyramidal dendritic growth of through the regulation of Signal Regulatory
Protein alpha (SIRPa) phosphorylation.

〇中村 史雄、若槻 実祐、瀧澤 光太郎、實木・高橋 葵
東京女子医科大・医We previously reported that protein tyrosine phosphatase delta (PTPd), one of type IIa receptor type protein
tyrosine phosphatases, mediates Sema3A-induced dendritic growth of cortical pyramidal neurons. However, its
endogenous substrates involved in cortical dendritic arborization have been yet identified. Phosphotyrosineproteome analysis of PTPd knockout brains revealed the hyperphosphorylation of Signal Regulatory Proteinalpha (SIRPa) at Tyr501 (Y501) residue. Immunohistochemistry with anti-phospho-Y501 SIRPa antibody
showed that olfactory epithelium, cortical II to V layers, thalamic nuclei and axon-bundles including corpus
callosum, fimbria, and pyramidal tract were hyperphosphorylated in PTPd knockout brains. Knockdown of
SIRPa by siRNA transfection or the overexpression of cytoplasmic deletion mutant of SIRPa suppressed
Sema3A-induced growth cone collapse response of mouse dorsal root ganglion neurons. Primary culture ofmouse cortical neurons revealed that Sema3A-stimulation induced the dephosphorylation of SIRPa in thedendritic growth cones of wild-type but not in those of PTPd knockouts. Overexpression of non-phosphorylated
SIRPa mutant Y501F in cultured cortical neurons attenuated Sema3A-induced dendritic growth. In utero
electroporation of SIRPa-Y501F to mouse brains showed that the apical dendrites of cortical layer II/III
pyramidal neurons were disoriented. Similar irregular projection of cortical apical dendrites was also observed in
PTPd knockout brains. These results suggest that PTPd may regulate the phosphorylation of SIRPa in corticaldendritic growth.

Copyright © The Japanese Pharmacological Society3-B-O06-3

一般演題（口頭）
2+

Development and application of label-free Ca Image Sensor to visualize
2+
extracellular Ca dynamics in hippocampus.

〇パラジュリ ビージェイ1,2、土井 英生3、繁冨 英治1,2、鈴木 秀明1,2、野田 俊彦3、高橋 一浩3、澤田 和明3、小泉 修一1,2
1

山梨大・院医・薬理、2山梨大・GLIAセンタ−、3豊橋技科大・電気・電子情報工学

Extracellular calcium ion ([Ca2+]o) change occurring during physiological and pathological conditions play
important roles in regulation of brain functions, but has received limited attention due to the lack of easy to use
device. Here, we show the development and application of highly selective calcium image sensor for the imaging
of glutamate-induced [Ca2+]o change in hippocampal slices. Using this sensor, we found that glutamate
decreased [Ca2+]o by more than 50 % with different spatiotemporal patterns. The glutamate-evoked decrease in[Ca2+]o was inhibited by a NMDA receptor antagonist D-AP5 but not AMPA receptor antagonist CNQX, andmimicked by NMDA. The spatial pattern of the glutamate-evoked [Ca2+]o decrease was associated with that of
distribution of NMDA receptors in the hippocampus. Moreover, using an ATP-imaging fluorescent probe, we
found that the stimulation with NMDA triggered increase in ATP release from astrocytes via either connexin
hemichannels or chloride channels. The NMDA-evoked ATP release was mediated by decrease in [Ca2+]obecause the astrocytic ATP release was mimicked by Ca2+-free medium. Taken together, using the newly
developed Ca2+ image sensor, we demonstrated that [Ca2+]o is dramatically decreased during excitatory synaptictransmission by glutamate, and [Ca2+]o decrease act as a signal that transmits neuronal excitation to astrocytes
via ATP release. The application of this Ca2+ sensor is expected to clarify the physiological andpathophysiological roles of [Ca2+]o , which have received limited attention so far.

Copyright © The Japanese Pharmacological Society3-B-O06-4

一般演題（口頭）

Analysis of synaptic structural changes induced by chronic social stress andtheir molecular mechanisms〇永井 裕崇1、永井 碧1、沼 知里1、大田 康平1、山下 朋美2、川島 祐介3、大野 伸彦4,5、片岡 洋祐6,7、新間 秀一8,9、清末 優子10、加藤 太朗2、曽我 朋義11、古屋敷 智之1
1

神戸大・院医、2住友ファーマ、3かずさDNA研究所、4自治医科大・医・組織、5生理研・超微形態、6理研―JEOL連携セ

ンター、7理研・生命機能科学研究セ・細胞機能評価、8大阪大・院工・生物工学、9大阪大・島津分析イノベーション協働研、10理研・生命機能科学研究セ・分子細胞動態研究チーム、11慶應義塾大・先端生命科学研

Excessive or chronic social stress induces emotional and cognitive disturbances and precipitates mental illness.
Altered neuronal morphology and functions in the medial prefrontal cortex (mPFC) underlie these behavioral
abnormalities. However, its subcellular mechanisms remain elusive. Here we examined ultrastructural andmulti-omics changes in the mPFC after social stress in mice. Social stress caused the loss of dendritic brancheswith morphological alterations of subcellular mitochondria and induced synaptic shrinkage selectively at thesynapses with mitochondria. Multi-omics and functional analyses revealed that social stress deteriorated
mitochondrial functions with altered mitochondrial proteome at synapses and dysregulated central metabolic
pathways in the mPFC. Molecular biological and pharmacological manipulation targeting central metabolism
and mitochondria attenuated the synaptic shrinkage and depression-related behaviors. These findings
demonstrate that chronic social stress alters the central metabolism at mPFC synapses, leading to neuronalpathology and depression-related behaviors.

Copyright © The Japanese Pharmacological Society3-B-O06-5

一般演題（口頭）

Analysis of neural circuit alterations caused by chronic social stress〇篠原 亮太1、奥田 裕己1、山口 真広2、小坂田 文隆2、古屋敷 智之1
神戸大・院医・薬理、2名古屋大・院創薬科学・細胞薬効解析

1

Stress caused by aversive and psychological stimuli induces sustained emotional and cognitive abnormalities andprecipitates the onset and relapse of symptoms in psychiatric illnesses, such as depression. We havedemonstrated that chronic social stress causes dendritic atrophy of pyramidal neurons in the medial prefrontal
cortex (mPFC) only in mice susceptible to chronic social stress, whereas acute social stress leads to dendritic
growth of these neurons concomitant with suppression of behavioral changes. However, it remains unknown
how stress-induced dendritic remodeling of mPFC pyramidal neurons affects neural circuits associated withdepressive behaviors. Here we utilized monosynaptic retrograde tracing with a G-deficient rabies viral vector to
identify neural projections to the mPFC that can be anatomically altered by chronic social stress. A G-deficientrabies viral vector encoding a red fluorescent protein (RFP) was unilaterally infused into the mPFC to visualize
neurons that directly input to the mPFC. Among 90 brain regions where RFP-positive cells were observed inmice subjected to chronic social stress, some particular regions showed a decrease or increase in RFP-positive
cells. This finding suggests that chronic social stress anatomically alters neural projections to the mPFC, leading
to sustained behavioral changes.

Copyright © The Japanese Pharmacological Society3-B-O06-6

一般演題（口頭）

Focus on the diabetic brain: Upregulation of orexin receptor and plasma orexin
level in obese diabetic rats

〇幸田 祐佳1、福石 信之2、松村 人志1
大阪医科薬科大・薬・薬物治療、2金城学院大・薬・薬理学

1Diabetes mellitus and brain toxicity are closely linked. Oxidative stress, obesity, insulin resistance, and glucose
toxicity can affect the brain. Orexin-A, also known as hypocretin-1, participates in many physiological processes
through its activated receptor. Orexin-A has been associated with feeding behavior, obesity, and pathogenesis of
Alzheimer's disease. We reported that high-dose thiamine in obese diabetic Otsuka Long‒Evans Tokushima
Fatty (OLETF) rats leads to reduced obesity and metabolic disorders. In addition, we found that plasma orexinA levels in OLETF rats can be modulated by thiamine supplementation under conditions of oxidative stress.
Herein, we focused on orexin-A in obese diabetic OLETF rats. At 58 weeks of age, the rats showed an increasein body weight and blood glucose levels. Plasma orexin-A was measured by ELISA and tended to be higher inobese diabetic OLETF rats than in non-obese diabetic control rats. We evaluated hypocretin receptor 1 (Hcrtr1,
also orexin-A receptor) gene expression in the brain of diabetic OLETF rats by reverse transcription-polymerase
chain reaction and found that diabetic OLETF rats exhibited higher orexin-A receptor gene expression in thebrain than controls. The results presented here are expected to provide a better understanding of the role oforexin-A and its contribution to diabetic brain.

Copyright © The Japanese Pharmacological Society3-B-O07-1

一般演題（口頭）

Changes in enteric cellular environment of the rotenone-induced parkinsonian
mice that reproduce central and enteric neurodegenerative features〇浅沼 幹人、宮崎 育子、進 浩太郎、都 明希、正井 加織、小林 壯太朗、津田 光希、小野 鈴香
岡山大・院医歯薬・脳神経機構

We previously established an animal model of Parkinsonʻs disease (PD) induced by chronic low-dose rotenone
as an environmental neurotoxic pesticide that reproducible central and enteric neurodegenerative features ofPD, and also found that rotenone-induced enteric neurodegeneration is caused by dysfunction of enteric glia
using primary cultured enteric cells. However, the mechanism of enteric neurodegeneration and inflammation
are still obscure. In this study, we examined changes in enteric cellular environment in the enteric epithelium
and myenteric plexus of the rotenone-induced PD model mice. Chronic subcutaneous administration with lowdose rotenone (2.5 mg/kg/day) for 4 weeks using an osmotic mini pump reduced the number of dopamine
neurons in the substantia nigra and the intestinal myenteric neurons and glial cells of mice. Furthermore, it
produced disruption of mucosal epithelial barrier and marked translocation of HMGB1 to the cytosol beside
nuclear membrane towards the apical lumen side. These results suggest that the rotenone-induced dysfunctionsof epithelial barrier and HMGB1 transportation are involved in the inflammatory reactions and dysfunction ofenteric glia and consequent enteric neurodegeneration.

Copyright © The Japanese Pharmacological Society3-B-O07-2

一般演題（口頭）

Role of microsomal prostaglandin E synthase-1 in hippocampal inflammation
after repetitive febrile seizures in mouse pups

〇松尾 由理1、友利 徳志1、竹本 陽祐1、植松 智2、審良 静男3、髙橋 達雄1
北陸大・薬・薬理、2大阪市立大・院医・ゲノム免疫、3大阪大・院医・免疫フロンティア研セ

1Febrile seizures (FSs) are the most common convulsive seizure in early childhood. About 30% of them are“complex”, with repetitive seizures, and related to adult temporal lobe epilepsy (TLE). In this study, weinvestigated the role of membrane-associated prostaglandin E synthase-1 (mPGES-1), an inducible terminal
enzyme for PGE2 synthesis, in hippocampal inflammation induced by repetitive FSs (RFSs) in mouse pups as amodel of complex FSs.
Wild-type (WT) and mPGES-1 knockout (ES1KO) mice at P9-11 were given intraperitoneal injections oflipopolysaccharide (100 µg/kg) and exposed to heat lamp to induce hyperthermia and FSs. The induction of FSs
was repeated twice at 4 h-interval (RFSs).
In WT mice, mPGES-1 mRNA was significantly up-regulated in hippocampus after RFSs. The production ofhippocampal PGE2 observed after RFSs in WT mice was completely absent in ES1KO mice. The seizure score
and increase in rectal temperature during the hyperthermia induction in ES1KO mice were slightly but
significantly lower than those in WT mice. The inductions of IL-1β, TNF-α and GFAP observed significantly
in WT mice were less in ES1KO mice even in which the seizure scores were almost the same level.
These results suggest that mPGES-1 contributes to inflammatory hyperthermia, convulsive events, glial
activation and production of inflammatory cytokines through PGE2 production in hippocampus. Thus, mPGES-1 may contribute to the complex FSs-induced adulthood TLE and may be a potential therapeutic target for thedevelopment of epilepsy after RFSs.

Copyright © The Japanese Pharmacological Society3-B-O07-3

一般演題（口頭）

Microvesicles are released from microglia in mouse models of both peripheral
acute inflammation and chronic skin inflammation.

〇坂井 謙斗1、小泉 修一1,2
1

山梨大・院医・山梨GLIAセンター、2山梨大・院医・薬理My research focuses on the relationship between peripheral inflammation and microglia. Lipopolysaccharide(LPS) is used to induce peripheral acute inflammation. It has been reported that LPS application induces
microvesicle (MV) release from microglia

and these MVs contain inflammatory cytokines such as IL-1b.

These data indicate that microglia detect peripheral inflammatory signals and react by releasing MVs. However,MVs released from microglia had not previously been observed
cranial windows into the heads of Iba1-GFP mice and performed. To observe microglial MVs, we madelive imaging using 2 photon-

microscopy. GFP+ MVs were observed 24h after LPS injection (1 mg/kg, ip), and the peak of the increase in
GFP+ MVs was at 48h after LPS injection. Because Iba1 is expressed in not only microglia but also somemacrophages, we treated mice with clodronate (33 mg/kg, ip) to deplete peripheral macrophages. Clodronate
treatment did not affect the increase of LPS-induced GFP+ MVs. This indicates that the GFP+ MVs werereleased from Iba1+ cells like microglia, but not macrophages. Interestingly, the GFP+ MVs were also observedin contact dermatitis model mice with chronic skin inflammation. These results provide a framework to study
the role of microglial MVs in peripheral inflammatory mouse models.

Copyright © The Japanese Pharmacological Society3-B-O07-4

一般演題（口頭）

Effects of KNT-127, a delta opioid receptor agonist, on non-REM sleep in mice.

〇岩井 孝志1、藤田 芽生1、渡辺 理紗子1、平山 重人2、尾山 実砂1、渡辺 俊1、藤井 秀明2、田辺 光男1
北里大・薬・薬理学、2北里大・薬・生命薬化学

1

Recently we have shown that delta opioid receptor agonists (DORs) shorten sleep time during the light period.
However, the impact of DOR agonists on sleep quality has not been determined. In the present study, weinvestigated the effects of the DOR agonist KNT-127 on delta wave power (1-4 Hz), which is an indicator of
non-rapid eye movement (REM) sleep depth. The vigilance states (e.g., wakefulness, REM and non-REM sleep)of the ddY-mice (6-10 weeks) were classified based on the hippocampal local field potential (LFP) and neckmuscle electromyogram. KNT-127 (10-30 mg/kg, i.p.) significantly decreased the mean REM and non-REM
sleep periods, and prolonged the mean wakefulness period during 5 hr after its injection. KNT-127 significantly
increased delta wave power during non-REM sleep compared to saline, and this effect was also observed at 3
mg/kg without the arousal. KNT-127 (3 mg/kg), when administered in the urethane-anesthetized mice,increased delta wave power, indicating that the action is not a rebound due to sleep suppression. Together, KNT-127 promotes deeper non-REM sleep independently of its arousal effects.

Copyright © The Japanese Pharmacological Society3-B-O07-5

一般演題（口頭）Propolis ameliorates cognitive decline in Alzheimer‘s disease model mice

〇稲垣 良1、山國 徹2,3、森口 茂樹1
1

東北大・院薬・医薬品開発研究センター、2東北大・院工・附属超臨界溶媒工学研究センター、3東北大・未来科学技術

共同研究センター

Propolis (Brazilian green propolis) is a chemically complex resinous substance that is expected to be beneficial
to therapy for Alzheimerʻs disease (AD). To reveal its beneficial effect on impaired cognition, we first performed
three memory-related behavior tasks in mice aged 4 and 12 months: Y-maze task, novel object recognition task,
and passive avoidance task. Oral dosages of 300-1000 mg/ kg once daily for 8 weeks, did significantly preventthe cognitive decline in the APP-KI mice aged 4 months, but not 12 months. Consistent with the observations
from behavioral tasks, impaired hippocampal long-term potentiation (LTP) was markedly ameliorated in theacute brain slices prepared from the mice that underwent the repeated propolis administration. In addition,
increased phosphorylation of CaMKII and AMPAR subunit (GluA1) was simultaneously observed in the CA1 ofthe mice. Similar to CaMKII activation, the propolis administration also increased CaMKIV and CREB
phosphorylation and BDNF production in the CA1 of the mice. Finally, we confirmed that the presence of 30
μg/ mL propolis significantly elevated intracellular Ca2+ concentration in Neuro2A cells. These findings suggestthat propolis is capable of rescuing the cognitive dysfunction via both upregulated activities of CaMKII and
CaMKIV in the CA1 of the APP-KI mice.

Copyright © The Japanese Pharmacological Society3-B-O08-1

一般演題（口頭）

Inhibitory role of small leucine-rich proteoglycans regulated by NOX1/NADPH
oxidase in cardiac fibrosis〇岩田 和実1、矢部 千尋2、楳村 敦詩1
京府医大・院医・病態分子薬理、2京都工場保健会総合医学研・臨床薬理学研究セ1

Cardiac fibrosis is a leading cause of heart failure, particularly heart failure with preserved ejection fraction(HFpEF). Currently, there is no effective treatment for HFpEF. We previously reported that doxorubicininduced cardiac fibrosis was suppressed in mice deficient in, a non-phagocytic isoform of superoxide-

producing NADPH oxidase. In this study, the role of NOX1 in the development of cardiac fibrosis wasinvestigated in cultured cells using a rat cardiomyoblast cell line H9c2 and cardiac fibroblasts isolated from adultmale mice. Increased proliferation was demonstrated when cardiac fibroblasts were exposed to homogenatesfrom wild-type H9c2. On the other hand, increased proliferation was significantly attenuated in cardiac
fibroblasts exposed to homogenates from

disrupted H9c2. In

disrupted H9c2 cells, the expression of

osteoglycin (Ogn), and podocan (Podn), which are small leucine-rich proteoglycans and known to regulate
cardiac remodeling, were up-regulated. When homogenates from-disrupted H9c2 with disruption of Ogn

or Podn exposed to cardiac fibroblasts, the proliferation of fibroblasts was significantly restored compared tothose exposed todisrupted H9c2. These findings suggest that NOX1 promotes cardiac fibrosis via down-

regulation of Ogn or Podn in cardiomyocytes.

Copyright © The Japanese Pharmacological Society3-B-O08-2

一般演題（口頭）

SGLT2 inhibitor reduces the inducibility and duration of atrial fibrillation in the
diet-induced obese mouse model.

〇澤野 達哉1、三明 淳一朗1、クルニアワン プリヨノ アグン1、岡村 昌宏2、友森 匠也2、高見 亜衣子2、長田 佳子1、今村 武史1
鳥取大・医・薬理学・薬物療法学分野、2鳥取大・医・循環器・内分泌代謝内科学分野

1

Background: Atrial fibrillation (AF) is the most common arrhythmia. AF is highly correlated with multiple risk
factors including heart failure, age, obesity, and type 2 diabetes. Among risk factors, the incidence in obesity is
increasing worldwide. Recently, it was reported that SGLT2 inhibitors reduced the incidence of atrial
fibrillation. However, it is unclear how the treatment with SGLT2 inhibitors has effects on vulnerability to AF. In
this study, we examined the effects on the inducibility and duration of AF by treatment with SGLT2 inhibitors indiet-induced obese mice.
Methods: Mice were fed a normal chow diet (NCD) or high-fat diet (HFD). Following diet-loading, we
randomly divided the animals into groups: NCD+vehicle, HFD+vehicle, and HFD+ SGLT2 treatments.
Induction of AF was performed by transesophageal atrial burst pacing. Furthermore, we evaluated cardiac
function, blood pressure, atrial fibrosis, and glucose tolerance at the end of the treatments.
Results: The results showed that HFD-fed mice increased the inducibility of AF compared to NCD mice. In
addition, treatment with the SGLT2 inhibitor in HFD-fed mice dose-dependently reduced the inducibility andduration of AF. There were no significant differences in cardiac function, blood pressure, and fibrosis among allgroups. Impairment of glucose tolerance in HFD-induced obesity was improved by treatment with the SGLT2
inhibitor.
Conclusion: Treatment with the SGLT2 inhibitor reduced the inducibility of AF and shortened the duration of
AF without affecting atrial structural remodeling, suggesting that the SGLT2 inhibitor effectively prevents AF in
obesity.

Copyright © The Japanese Pharmacological Society3-B-O08-3

一般演題（口頭）

Evaluation of antiarrhythmic agents for catecholaminergic polymorphic
ventricular tachycardia (CPVT) using multiple lines of RyR2-mutant mousemodels〇呉林 なごみ1、児玉 昌美1、村山 尚1、杉原 匡美2、石井 光一郎1、岡部 雄太3、村越 伸行3、三浦 綾4、西尾 元4、井上-上野 由紀子5、井上 高良5、野口 悟5、中村 衣里6、金井 富三夫6、多田 昇弘6、櫻井 隆1
1

順天堂大・医・薬理、2筑波大・医・循内、3順天堂大・医・臨床検査医学、4兵庫医大・医・法医、5国立精神・神経医療研

究セ、6順天堂大・院医・疾患モデル研究セ

Gain-of-function mutations in type 2 ryanodine receptors (RyR2) are known to cause severe arrhythmias suchas catecholaminergic polymorphic ventricular tachycardia (CPVT). Conventional antiarrhythmic drugs are
sometimes insufficient to suppress these arrhythmias. To seek more effective drugs, we aimed to provide a basis
for quantitative evaluation of the two important effects of antiarrhythmic drugs, i.e., prevention andtreatment/stopping of arrhythmia, using multiple RyR2 mutant mouse lines (R420W, I4093V and K4750Q)
with varying degrees of enhanced Ca2+ release activity. Short term ECG was recorded from the limb leads underisoflurane anesthesia, and arrhythmia was induced by administration of adrenaline alone or adrenaline/caffeine
mixture. For monitoring basal arrhythmia during everyday activities, long range ECG was recorded by telemetry
system. The R420W mice having moderately activated channels showed little basal arrhythmia but exhibitedsevere ventricular arrhythmias by adrenaline/caffeine induction. On the contrary, the RyR2-I4093V andK4750Q strains having highly activated channels showed frequent basal arrhythmia without adrenergic
induction. Na channel blockers, Ca channel blockers and beta blockers suppressed the induced arrhythmia andbasal arrhythmia to varying degrees. Interestingly, the preventive effect and the stopping effect seemed to differ
depending on their mechanism of action. The usage of multiple lines of mice with different degrees of activity
are useful for the evaluation of therapies for CPVT.

Copyright © The Japanese Pharmacological Society3-B-O08-4

一般演題（口頭）

Methylglyoxal-induced enhancement of uridine diphosphate-mediatedcontraction in rat femoral artery was due to activation of p38 MAPK and Syktyrosine kinase〇松本 貴之、吉岡 円、野上 あかね、工藤 美代、長田 友恵、小笠原 悠、田口 夏芽、遠藤 彩香、中村 圭吾、山田 愛
子、田口 久美子、小林 恒雄
星薬科大

Although abnormalities of function in femoral artery, including aberrant vascular reactivity to various vasoactive
mediators, are common in many chronic disorders including hypertension and diabetes, their inducible and/or
progressive factors remain unclear. On the other hand, methylglyoxal (MGO), a highly reactive dicarbonylcompound, has been implicated in the pathogenesis of several chronic disorders. Although our previous studies
demonstrated that uridine 5ʻ-diphosphate (UDP)-induced contraction in the femoral artery is increased in
hypertensive rat model, the direct relationship between MGO and UDP-mediated contraction is currently
unknown in rat femoral artery. We therefore investigated the acute effect of MGO (4.2 × 10−4 M for 60 min) on
UDP-induced contraction in the rat femoral artery. MGO amplified the UDP-induced contraction in the Wistar
rat femoral artery. This augmented response was not abolished in all conditions, including nitric oxide synthaseinhibition by L-NNA (10−4 M), cyclooxygenase inhibition by indomethacin (10−5 M), or endothelial
denudation. Moreover, in the endothelium-denuded arteries, the p38 mitogen-activated protein kinase (MAPK)
inhibitor SB203580 (10−5 M) reduced the UDP-induced contraction in both control and MGO-treated groups,
while MGO enhanced the p38 MAPK activation regardless of the UDP presence. Moreover, in theendothelium-denuded arteries, the Syk tyrosine kinase inhibitor piceatannol (10−5 M) reduced the UDPinduced contraction in both control and MGO-treated groups. These results suggest that MGO enhances UDPinduced contraction in rat femoral arteries and that this enhancement may be partly due to increases in theactivities of Syk tyrosine kinase and p38 MAPK in femoral arterial smooth muscle.

Copyright © The Japanese Pharmacological Society3-B-O08-5

一般演題（口頭）

Anti-spasmolytic effects of BAY 60-2770, a soluble guanylate cyclase activator, in
isolated coronary arteries〇田和 正志、中川 恵輔、大喜多 守
大阪医科薬科大学・薬・病態分子薬理This study investigated whether soluble guanylate cyclase (sGC) activators (activating the heme-oxidized andheme-free sGC) have potential as therapeutic drugs for coronary artery spasm. Isolated canine and porcine
coronary arteries were suspended in organ chambers for isometric tension recording. The contractile responsesof canine coronary arteries to potassium chloride (3×10-2 M), endothelin-1 (3×10-11 to 3×10-8 M),
prostaglandin F2α (10-8 to 10-5 M), and 5-hydroxytryptamine (10-9 to 10-6 M) were suppressed by previous
exposure to the sGC activator BAY 60-2770 (10-10, 10-9 and 10-8 M). In porcine coronary arteries, the addition of
BAY 60-2770 (10-10, 10-9 and 10-8 M) concentration-dependently prolonged the cycle length of 3,4diaminopyridine (10-2 M)-induced phasic contractions and reduced the peak tension. As for vessel-sizedependent difference in vasoreactivity, BAY 60-2770 (10-12 to 10-7 M) caused a greater relaxation of porcine
coronary arteries precontracted with endothelin-1 (3×10-8 M) in small arteries (#9 in AHA classification) thanin large arteries (#6 in AHA classification). These findings suggest that sGC activators are beneficial for thetreatment of vasospastic angina. In addition, anti-spasmolytic efficacy of sGC activators may be expected to beobserved even in microvascular angina.

Copyright © The Japanese Pharmacological Society3-B-O08-6

一般演題（口頭）Electropharmacological analysis of vernakalant as an anti-atrial fibrillatory drugusing the isoflurane-anesthetized beagle dogs and the rat aorta〇後藤 愛1、神林 隆一1、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

Vernakalant was approved as an anti-atrial fibrillatory drug by EMA and Health Canada but not by
FDA due to its cardiovascular adverse events including bradycardia and hypotension. We characterized its
electropharmacological profile

with the isoflurane-anesthetized beagle dogs andwith the rat

aorta.
Vernakalant (0.3 and 3 mg/kg/10 min) was intravenously administered to the dogs in theabsence (n=5) and presence (n=4) of α-adrenoceptor blocker phentolamine. Next, thevascular effectof vernakalant on the rat aorta was assessed by its cumulative application in concentrations of 0.001-100μmol/L (n=13 preparations).
Vernakalant suppressed the sinus automaticity, ventricular contractility and atrioventricular nodal aswell as intraventricular conduction, whereas it increased the total peripheral vascular resistance, preload to theleft ventricle and mean blood pressure. It delayed the ventricular repolarization in a reverse frequencydependent manner; the extent of prolongation of early and late repolarization was similar. It also prolonged theatrial and ventricular effective refractory period similarly. Pretreatment of phentolamine hardly affected thoseresults. Meanwhile, vernakalant did not induce the contraction of aorta.

Vernakalant exerted α-adrenodceptor-independent vasoconstrictor action only

. It

also showed electrophysiological effects on the atria and ventricles to a similar extent, which resembles thoseof-sotalol and bepridil.

Copyright © The Japanese Pharmacological Society3-B-O09-1

一般演題（口頭）Runx1 is upregulated by STAT3 and promotes cell proliferation in neonatal rat
cardiomyocytes.

〇鈴木 翔大1、亀谷 祐介1、梅田 綾香1、西中 康介1、田中 翔大1、尾花 理徳1,2、岡田 欣晃1、藤尾 慈1,2
大阪大・院薬・臨床薬効解析学分野、2大阪大・先導的学際研究機構生命科学融合フロンティア研究部

1【Background】
Mammalian cardiomyocytes (CMs) retain proliferative capacity shortly after birth, though these cells aredifferentiated with contractile activity. By using Runx1 as a dedifferentiation marker, recent studies havedemonstrated that CMs are dedifferentiated prior to cell proliferation; however, biological significance of Runx1
remains to be fully elucidated. The aim of this research is to clarify the regulatory mechanisms of Runx1
expression and its biological functions in CM proliferation.
【Method/Result】
CMs were prepared from neonatal rats. Cell proliferative activity was estimated by immune-fluorescent
microscopic analysis with anti-Ki-67 antibody. CM exhibited proliferative activity in response to fetal bovine
serum (FBS). Previously, since we demonstrated that STAT3 plays an important role in CM proliferation, theeffects of STAT3 on CM proliferation was analyzed by using siRNA. STAT3 knockdown reduced the frequencyof Ki-67+ CM, accompanied by the decrease in Runx1 expression. Importantly, Runx1 knockdown also
suppressed CM proliferation in response to FBS.
【Conclusion】
Runx1 expression is regulated by STAT3, and promotes CM proliferation, indicating the functional importanceof Runx1 in cardiac proliferation.

Copyright © The Japanese Pharmacological Society3-B-O09-2

一般演題（口頭）

2,5-Dimethylcelecoxib prevents cardiac remodeling associated with myocardialischemia/reperfusion injury in mice

〇石兼 真1、幾島 栄悟1、岸上 赳大1、井川 和宣2、友岡 克彦2、有岡 将基1、高橋 富美1
1

産業医科大・医・薬理、2九州大・先導物質化学研・分子集積化学

Myocardial ischemia/reperfusion (MI/R) injury leads to aggravated cardiac remodeling and heart
failure. Previously, we reported that 2,5-dimethylelcelecoxib (DMC), a derivative of celecoxib without
cyclooxygenase-2 inhibition, prevents cardiac remodeling in a non-ischemic cardiac fibrosis model. In this study,
we examined whether DMC inhibited myocardial remodeling associated with MI/R injury. The left anterior
descending coronary artery was ligatured for 0.5 hours and subsequently subjected to reperfusion for MI/R
injury in male C57 BL/6 mice. Vehicle or DMC was administered orally (DMC: 150 mg/kg) immediately after
awakening and followed by feeding (DMC: 1000 ppm). Echocardiographic evaluation showed significant
improvement of left ventricular ejection fraction in the DMC-treated group compared to the Vehicle-treatedgroup at 1-4 weeks after MI/R injury. In MI/R-injured hearts, protein expression of alpha-smooth muscle actin(myofibroblast marker) was significantly reduced by DMC treatment, as were mRNA expressions of fibronectin,
connective tissue growth factor, and matrix metalloproteinase-9, 3 days after injury. Masson trichrome staining
indicated that DMC significantly reduced cardiac fibrosis area 4 weeks after MI/R injury. This study revealed
that DMC decreased myofibroblast appearance, and suppressed fibrosis and cardiac dysfunction associated with
MI/R injury. DMC might be useful for preventing the development of heart failure associated with reperfusion
therapy for acute myocardial infarction.

Copyright © The Japanese Pharmacological Society3-B-O09-3

一般演題（口頭）

Establishment of a novel method for analysis of biological functions of myeloid
cell subpopulation in cardiovascular diseases using TRECK system.

〇横田 洸成1、冨松 聖史1、小南 春祐1、梶浦 僚太1、田中 翔大1、尾花 理徳1,2、岡田 欣晃1、藤尾 慈1,2
大阪大・院薬・臨床薬効解析学分野、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］Though the importance of myeloid cells in the cardiac remodeling after myocardial infarction(MI) is widely accepted, it remains to be fully elucidated how myeloid cells regulate post-infarct inflammation,
at least partially, because subpopulation-specific cell knock-out methods are not available.
［Methods and Results］We generated transgenic mice expressing diphtheria toxin receptor (DTR)/GFP
fusion protein under the control of CD11b promoter in a Cre recombinase-expressing cell-specific manner(CD11b-DTR TG mice). Double TG mice (DTG mice) were generated by crossing CD11b-DTR TG mice with
LysM-Cre mice that express Cre recombinase preferentially in monocytes/macrophages. The MI model was
created in DTG mice by ligation of the left anterior descending branch. Flow cytometry analysis revealed thatmonocytes were labeled with GFP in the peripheral blood 4 days after MI. Consistently, immunofluorescent
microscopic analysis showed that GFP⁺ cells inﬁltrated into the infarcted heart. Importantly, the administrationof diphtheria toxin resulted in the depletion of GFP⁺ cells in peripheral blood and post-infarct myocardium.
［Conclusion］CD11b-DTR TG mice are useful for labeling and/or depleting subpopulation of myeloid cells in
MI model.

Copyright © The Japanese Pharmacological Society3-B-O09-4

一般演題（口頭）

Gas phase extract of mainstream smoke derived from heated tobacco productscauses iron-dependent, ferroptosis-independent cell injury in vascularendothelial cells.

〇堀之内 孝広1、三輪 聡一2
1

北海道大・院医・細胞薬理、2公立豊岡病院

Ferroptosis is defined as an iron-dependent regulated necrosis that is caused by massive lipid peroxidationmediated cell membrane damage. The present study examined whether ferroptosis is involved in endothelial
dysfunction by nicotine- and tar-free cigarette smoke extract (CSE) prepared from heated tobacco products(HTPs). The CSE of HTPs (Ploom X, IQOS 3, and IQOS ILUMA) and a combustion cigarette (1R6F) was
prepared according to Health Canada Intense smoking method (55 mL puff volume, 2 sec puff duration, and 1
puff every 30 sec) using an analytical vaping machine LM5E (Borgwaldt KC GmbH). The cytotoxicity of CSE of
HTPs and 1R6F to human umbilical vein endothelial EA.hy926 cells was evaluated by measuring mitochondrial
metabolic activity and lactate dehydrogenase (LDH) leakage. CSE from cigarettes except for Ploom X, anderastin, a ferroptosis inducer by inhibiting cystine-glutamate exchange transporter (system XC-), triggered adecrease in mitochondrial metabolic activity and an increase in LDH leakage. The cytotoxic effects of CSE of
IQOS 3 and 1R6F were reduced by an iron chelator deferoxamine mesylate (DFO), but not by a ferroptosis
inhibitor UAMC-3203, which scavenges lipid reactive oxygen species (ROS). On the other hand, erastin
cytotoxicity was inhibited by both DFO and UAMC-3203. These results suggest that erastin-induced, irondependent ferroptosis leads to cell damage characterized by a decrease in mitochondrial metabolic activity andan increase in LDH leakage, in EA.hy926 cells. CSE of IQOS 3 and 1R6F causes iron-dependent mitochondrialand cell membrane damage, both of which are independent of lipid peroxidation by ROS.

Copyright © The Japanese Pharmacological Society3-B-O09-5

一般演題（口頭）

Glucocorticoid first increases urinary sodium excretion and urine volume,
leading to skin sodium and water loss in mice

〇北田 研人、中野 大介、西山 成
香川大・医・薬理学

We previously revealed that glucocorticoids such as cortisol and cortisone may also regulate sodium and fluidbalance in healthy subjects. We also reported that dexamethasone administration to mice initially increased
urinary sodium excretion and urine volume and reduced skin sodium and water content in the early phase ofadministration and that chronic dexamethasone injection only decreased skin sodium and water content.
However, it remains to be clarified which organ, kidney or skin, is responsible for the initial sodium and water
loss at initial dexamethasone injection. In the present study, we examined the effects of dexamethasone on skinsodium and water content in bilateral nephrectomized mice. In the sham-operated group, dexamethasone (1mg/kg/day, s.c.) significantly increased urinary sodium excretion and urine volume and decreased skin sodium
and water content 24 hours after the injection. Dexamethasone did not affect plasma sodium concentration andosmolarity. In bilateral nephrectomy groups, dexamethasone did not alter skin sodium and water content. Thesefindings suggest that glucocorticoid originally increases urinary sodium excretion and urine volume, which
decreases skin sodium and water loss to compensate for renal sodium and water loss. In order to elucidate themechanisms of sodium and fluid homeostasis, it may be necessary to examine skin and glucocorticoids inaddition to the known hormones and kidney systems.

Copyright © The Japanese Pharmacological Society3-B-O10-1

一般演題（口頭）

Aquaporin-5 in airway epithelial cells enhance LPS-induced cytokine expressionin vivo experiment.

〇石井 慎也1、岡谷 拓海2、内山 雄太2、礒濱 洋一郎2、村上 一仁2
東京理科大・院薬・応用薬理、2東京理科大・薬・応用薬理1

Copyright © The Japanese Pharmacological Society3-B-O10-2

一般演題（口頭）

Copyright © The Japanese Pharmacological Society3-B-O10-3

一般演題（口頭）

Effect of local IL-10 replacement therapy on severe asthma in mice

〇松田 将也、大森 美侑、北谷 和之、奈邉 健
摂南大・薬・薬効薬理

Subgroups of severe asthma patients with marked increases in sputum eosinophils and neutrophils areinsensitive to corticosteroids. Exogenous administration of IL-10 negatively regulates both migration ofeosinophilic and neutrophilic into tissues. This study evaluated whether intratracheal IL-10 administration
suppresses asthmatic responses in a severe model of mice. Ovalbumin (OVA)-sensitized mice wereintratracheally challenged with OVA. Dexamethasone (DEX, 1 mg/kg, intraperitoneal) or IL-10 (25 ng/mouse,
intratracheal) was administered during the challenges. The number of leukocytes, expressions of adhesion
molecules and IL-10 receptor, and development of airway hyperresponsiveness (AHR) were evaluated after thechallenges. Although DEX hardly suppressed the development of AHR, the infiltration of eosinophils andneutrophils, and the development of AHR were significantly inhibited by intratracheal IL-10 administration.
Moreover, IL-10 administration markedly decreased the numbers of ICAM-1+ and VCAM-1+ pulmonary
vascular endothelial cells, which express IL-10 receptor 1. IL-10 could suppress eosinophil and neutrophil
infiltration by inhibiting the proliferation of ICAM-1+ and VCAM-1+ pulmonary vascular endothelial cells,
resulting in inhibition of AHR in severe asthmatic mice.

Copyright © The Japanese Pharmacological Society3-B-O10-4

一般演題（口頭）

Role of histone ubiquitination in SARS-CoV2 and influenza virus infection〇衣笠 泰葉、星崎 みどり、Llamas Covarrubias Mara、今井 由美子
医薬基盤研・感染病態制御ワクチンPJ

Virus infection may affect the epigenetic regulations in host cells, including post-translational histone
modifications. Ubiquitination of histone H2B, has been reported to be involved in transcription activation.
However, it remains unknown the role of histone ubiquitination in the pathology of virus infection (e.g.
influenza virus, SARS-CoV2). CNOT4 that is a component of the CCR4-NOT complex has a ubiquitintransferase activity at the RING domain (L16). Here we show that CNOT4 is responsible for histone H2Bubiquitination in the host cells, which was linked to H3K4 methylation. Upon influenza virus or SARS-CoV2
virus infection CNOT4 interacted to virus protein, resulting in the loss of H2B ubiquitination and H3K4
methylation, which suppress interferon-related gene expression. The cells with a ubiquitination activity site of
L16 of CNOT4, have increased virus replication. These results suggest that the CNOT4 is involved in the virus
replication through histone H2B ubiquitination.

Copyright © The Japanese Pharmacological Society3-B-O10-5

一般演題（口頭）

Chemokine CCL28 suppresses liver fibrosis in a mouse model of carbon
tetrachloride-induced chronic hepatitis〇原 雄大、細川 隼矢、奥村 遼平、松尾 一彦、中山 隆志
近畿大・薬・化学療法

The chemokine CCL28 is mainly expressed in mucosal tissues such as colon and salivary gland, and migrates
IgA-secreting cells via its receptor CCR10. Because IgA protects mucosal tissues from pathogenic
microorganisms, CCL28 is thought to play an important role in mucosal immunity. However, CCL28 has beenshown to be also expressed in non-mucosal tissues. Although some previous studies reported that CCL28 wasincreased in liver of patients with chronic liver disease, the detailed involvement in the pathology remains
unclear. In this study, we examined the influence of CCL28 deficiency on carbon tetrachloride (CCl4)-induced
chronic hepatitis in mice. Chronic treatment with CCl4 increased CCL28 expression levels in the liver. CCL28deficient mice showed increased serum ALT levels and fibrotic areas of the liver. CCl4 treatment also increased
IgA-secreting plasma cells (PC), which expressed CCR10, in the liver of wild-type mice, but not CCL28deficient mice. The hepatic IgA-secreting PC, but not IgA-negative PC, expressed IL-10, FasL, and PD-L1.
Furthermore, CCL28-deficient mice showed decreased apoptosis and activity of hepatic stellate cells which are a
key player in the progression of liver fibrosis. These findings suggest that CCL28 would suppress the pathology
of chronic hepatitis through the migration of IgA-secreting PC.

Copyright © The Japanese Pharmacological Society3-B-O10-6

一般演題（口頭）

Intraperitoneal injection of the chemokine CX3CL1 improves aged recognition
memory〇武井 義則、後藤 愛、神林 隆一、中瀬古（泉） 寛子、杉山 篤
東邦大・医・薬理学講座

Aging is associated with a progressive decline in cognitive function. While obesity accelerates aging process ofthe brain,physical activity preserves and improves cognitive performance in the elderly. The expression of achemokine CX3CL1 has been reported to be increased in both conditions. CX3CL1 is known to chemoattract T
cells and monocytes. Moreover, it promotes survival of neurons in the brain and b-cells in the pancreas. In
adipose tissues, it is expressed in adipose cells and attenuates effects of obesity-induced chronic
inflammation. However, the role of the lifestyle-induced CX3CL1 expression in cognitive aging is unknown.
Here, we administered CX3CL1 into the peritoneal cavity of aged mice (15-16 months old) to investigate its
impact on the aging process of cognition. In the hippocampus, CX3CL1 increased the number of Type-2 neuralstem cells and promoted brain-derived neurotrophic factor (BDNF) expression. This treatment, furthermore,
improved novel object recognition memory impaired with advancing age. Intraperitoneal transplantation of
peritoneal cells from CX3CL1-treated aged mice improved novel object recognition memory in recipient aged
mice. Vagotomy inhibited the CX3CL1-induced increase in BDNF expression. Thus, our results demonstratethat a novel connection among peritoneal cells, the vagal nerve and the hippocampus can reverse the ageassociated decline in recognition memory.

Copyright © The Japanese Pharmacological Society3-B-O11-1

一般演題（口頭）

IL-34 inhibition attenuates renal fibrosis induced by unilateral ureteralobstruction in mice

〇和田 幸寛1、本田 浩一2、伊藤 義也1、細野 加奈子1、畑中 畑中1、天野 英樹1
1

北里大・医学部薬理学講座、2昭和大・医・内科学講座腎臓内科学部門

Interleukin (IL)-34, a macrophage (Mø) mediator, is expressed by tubular epithelial cells.
However, the influence of IL-34 on tubulointerstitial fibrosis remains to be fully elucidated. We investigated theeffect of IL-34 on renal fibrosis caused by unilateral ureteral obstruction (UUO).

10-

week-old male C57BL/6 (B6) mice (n=16) were induced UUO. Groups of animals were given either antimouse IL-34 antibody (UUO+anti-IL-34 Ab, 400 ng/kg, n=8) or vehicle (UUO+V, equal volume of saline,n=8) daily by intraperitoneal injection. Four age-matched male B6 mice received sham operation as control. All
Compared to the control, the UUO+V mice exhibited remarkablemice were sacrificed on day 10.

intrarenal expressions of IL-34 and its two receptors (cFMS and PTP-ζ), which were significantly suppressedby anti-IL-34 Ab treatment. Compared to the UUO+V mice, tubular injury and sirius red positive area weresignificantly attenuated in the UUO+anti-IL-34 Ab mice. Treatment with anti-IL-34 Ab significantly
suppressed the number of F4/80+ Mø and α-SMA+ myofibroblast in damaged kidneys of UUO. The renal
cortical transcript levels of TGF-β, COL-1, TNF-α, IL-6, MCP-1/CCL2, and MIP-1/CCL3 were significantly
lower in the UUO+anti-IL-34 Ab mice.

Elevated IL-34 expression was related to renal fibrosis.

Inhibition of IL-34 with neutralizing Ab suppressed expressions of inflammatory cytokines and fibrogenetic
genes via reducing the Mø infiltration, which might lead to attenuate the development of renal fibrosis.

Copyright © The Japanese Pharmacological Society3-B-O11-2

一般演題（口頭）

Bipolar effect of cannabinoid CB2 receptors to peripheral neuroinflammation〇野崎 千尋1、Zimmer Andreas2、柴田 重信31

早稲田大・国際理工学センター・Major in Bioscience、2University of Bonn・Institute of Molecular Psychiatry、3早稲田

大・理工・電気・情報生命工学科

It is widely known that cannabinoid type 2 (CB2) receptor deficiency enhances neuroinflammation and pain
development in the animal model of nerve injury-evoked neuropathic pain. We previously proposed theupregulated leptin signaling at the peripheral nerve as one of the underlying molecular mechanism, as nerveinjured CB2 receptor knockouts (CB2-KO) displayed robust upregulation of leptin receptors in both injured
and non-injured nerve tissue. Those leptin receptors seemed to be expressed on the macrophages which is
recruited to the nerve by the tissue injury, indicating the infiltration of leptin receptor-expressing macrophages.
Thus, Due to these past results we also hypothesized that lack of CB2 receptor might also enhance the high fat
diet (HFD)-induced peripheral neuroinflammation. However, surprisingly, CB2-KOs showed the significant
resistance to the HFD-induced neuroinflammation. Namely, 5-week feeding of HFD induced substantial
hypersensitivity in WT mice, while tactile sensitivity of HFD-fed CB2-KO remained intact. In the same animals,
we further found the significant upregulation of infiltrated macrophages and chemokine receptor CXCR4
expression in HFD-fed WT animals, but not in either HFD-fed CB2 knockout mice or standard fat diet (SFD)fed WT and CB2-KO controls. Based on these results, we will propose that CB2 receptors might have thebipolar regulatory role to chemokine receptor-mediated inflammatory response, which in the end enhance orinhibit the development of neuroinflammation depending on its cause.

Copyright © The Japanese Pharmacological Society3-B-O11-3

一般演題（口頭）

Effect of berberine on dermatitis and itch-related responses in mice with atopylike dermatitis〇安東 嗣修1、田渕 圭章2、清水 忠道3
金城学院大・薬・病態薬理、2富山大・遺伝子実験施設、3富山大・医・皮膚科学

1Atopic dermatitis (AD) is a chronic skin disease with severe inflammation and pruritus. Traditional Kampo
medicine Orengedokuto improves AD symptom in human patients and atopy-like symptoms in AD mousemodel. Berberine is a major component of Orengedokuto. This study investigated the effects and molecularmechanisms of berberine on AD-like symptoms in mice. In NC/Nga mice with atopy-like dermatitis (dermatitismice), intermittent oral administrations of berberine inhibited skin symptom, itching, cutaneous infiltration ofeosinophils and mast cells, and the cutaneous expression of eotaxin, macrophage migration inhibitory factor(MIF) and IL-4. Berberine also inhibited both IL-4/MIF-induced eotaxin in fibroblasts and allergen-induced
MIF and IL-4 in mast cells. In mast cells, the GeneChip® microarray analysis showed that antigen increased theexpression of EIF3F and MALT1, inhibited by berberine. The regulation of these factors by siRNAs for themshowed antigen-induced the expression of MIF and IL-4. These results suggest that berberine inhibits AD-like
symptoms through at least downregulation of EIF3F and MALT1 in mast cells.

Copyright © The Japanese Pharmacological Society3-B-O11-4

一般演題（口頭）

Acquired oral immune tolerance was overridden by exposure of food antigen via
skin to use murine food allergy models〇山下 弘高1、伊波 幸紀1、田中 宏幸2,3、稲垣 直樹4、筒井 正人1
琉球大・院医・薬理、2岐阜薬科大・院薬・免疫生物、3岐阜大・院連合創薬、4岐阜医療科学大・薬・薬理1

Oral tolerance (OT) was immune regulatory system for foods. Food allergy (FA) patients are induced allergy byinhibition of acquiring OT or overriding OT. Recently, it has been hypothesized that immune tolerance is
induced for foods taken orally and that allergy is caused by exposure via skin. In this study, we tried to override
OT through exposure of food antigen to skin. We established three murine FA models to analyze therelationship between OT and percutaneous sensitization. IP model: Mice were injected ovalbumin (OVA)intraperitoneally (IP) followed by induction of food allergy. EC model: Mice were pasted a filter paper
containing OVA on shaved back skin for epicutaneous sensitization (EC). ID model: Mice were injected OVA
intradermally (ID) for sensitization via skin. OT was induced by oral OVA treatment before the sensitization ineach model. FA was estimated by drop of body temperature, diarrhea, and OVA-specific IgE level in plasma. In
all FA models, we confirmed FA symptoms involving elevation of OVA-specific IgE. OT induction inhibited theincreasing in IgE level and suppressed FA in IP model. In EC model with OT induction, we detected increasing
in IgE level. In ID model with OT induction, we confirmed FA symptoms and increasing in IgE level. The data
indicated that exposure of OVA via skin could override OT.

Copyright © The Japanese Pharmacological Society3-B-O11-5

一般演題（口頭）

Effect of Japanese cedar pollen sublingual immunotherapy on allergic rhinitis
symptoms during the Japanese cypress pollen dispersal period〇土井-大橋 雅津代1、松原 弘季1、大久保 公裕2、岡本 美孝3、前川 祐理子1
鳥居薬品（株）、2日本医科大学大学院医学研究科、3ちば労災病院

1

Background: The clinical efficacy of Japanese cedar (JC) pollen SLIT tablets for allergic symptoms during
Japanese cypress (JCY) pollen dispersal has been controversial in actual clinical settings. JC and JCY bothbelong to thefamily and the major allergens of the two species have strong amino acid homology.

The efficacy of JC pollen SLIT tablets against seasonal allergic rhinitis during the JC pollen and JCY pollen
dispersal periods was investigated.
Methods: A

analysis was conducted in a phase II/III study (JapicCTI no. 142579). Patients with JC

pollinosis (aged 5-64 years) were included (placebo n=159, 5000 JAU n=158). Patients in the active 5000 JAU
treatment group was treated with JC SLIT tablet daily by self-administration for the duration of the trial.
Clinical efficacy was evaluated by the total nasal symptom and medication score (TNSMS) during the peak
symptom periods of each pollen season over 3 years.
Results: The daily average TNSMS in the 5000 JAU group was consistently lower than in the placebo groupduring both the JC and JCY pollen dispersal periods in all three seasons. The larger reduction in TNSMS in the5000 JAU group compared with the placebo group was observed in a treatment duration-dependent manner.
Conclusions: JC SLIT tablet treatment showed sustained clinical efficacy on allergic symptoms during 3
consecutive JCY pollen seasons. Further studies are required to examine the immunological responses andextent of antibody cross-reactivity towards the homologous major allergens from different
species including JC and JCY.

Copyright © The Japanese Pharmacological Society

family4-B-O12-1

一般演題（口頭）

Involvement of inflammatory cells in the dorsal root ganglion in oxaliplatininduced neuropathic pain

〇大澤 匡弘1、岩城 杏奈1、梅澤 直樹2、樋口 恒彦2、杉山 洋介4、祖父江 和哉3、粂 和彦1
名古屋市立大・院薬、2名古屋市立大・院薬・精密有機反応学分野、3名古屋市立大・院医・麻酔科学・集中治療医学分

1野、4名古屋市立大・院医・臨床薬剤学分野

Oxaliplatin is a platinum-based chemotherapeutic agent treating colorectal and gastric cancer. The mostcommon dose-limiting adverse event of oxaliplatin treatment is chronic neuropathic pain, while the detailed
mechanisms remain unknown. The present study investigated the molecular mechanisms of microglial
regulation of the oxaliplatin-induced neuropathic pain. Intrathecal (I.t.) treatment with inducible nitric oxide
synthetase (iNOS) inhibitor 1400W attenuated the oxaliplatin-induced cold and mechanical hypersensitivity. In
addition, pharmacological and chemogenetical inhibition of macrophage/microglia also attenuated theoxaliplatin-induced cold and mechanical hyperalgesia. I.t. treatment with STAT3 inhibitor stattic attenuated theoxaliplatin-indued cold and mechanical hypersensitivity. Oxaliplatin induced the increased phosphorylation ofneural STAT3 in the dorsal root ganglion (DRG), which was attenuated by i.t. pretreatment with 1400W. Since
STAT3 phosphorylation was regulated by PTEN, effects of oxaliplatin on the PTEN activity in the DRG were
examined. PTEN expression was decreased by oxaliplatin treatment. Our present study suggested thatoxaliplatin induces neural STAT3 activation through the iNOS induction by macrophage/microglial activationin the DRG, which resulted in the oxaliplatin-induced neuropathic pain.

Copyright © The Japanese Pharmacological Society4-B-O12-2

一般演題（口頭）

Extrinsic ribosome stimuli triggers development of glioblastoma stem like cells〇城野 博史1,3、白川 裕貴2、三宅 俊介1,3、金丸 歩美3、米丸 興3、内野 翔太3、秀 拓一郎4、武笠 晃丈5、太田 訓正6、齋藤 秀之1,3
熊本大・大学病院・薬剤部、2順天堂大・院医、3熊本大・大学院薬学教育部、4北里大・大学病院、5熊本大・院生命科

1学、6九州大・基幹教育院: Although glioblastoma (GBM) stem like cells (GSCs), which exhibit chemo-radio resistance andrecurrence, are key prognostic factors in GBM patients, molecular mechanisms of GSCs development are largely
unknown. Recent studies revealed that extrinsic ribosome incorporation into somatic cells played key roles incell reprogramming process towards stem cell properties. In this study, we sought to elucidate the mechanisms
underlying GSCs development by focusing extrinsic ribosome incorporation into GBM cells.
: The ribosome incorporation into GBM cells significantly increased ribosome induced cancer cellspheroid (RICCS) formation, and showed the stem like cell characters. In RICCS, phosphorylation and protein
expression of RPS6, an intrinsic ribosomal protein, and STAT3 phosphorylation were involved in regulating cellspheroids formation. Interestingly, glioma-derived extrinsic ribosome also promoted GBM-RICCS formation
through the intrinsic RPS6 phosphorylation. Moreover, in glioma patients, RPS6 phosphorylation was observedin higher grade glioma tissues, and predominantly up-regulated in GSCs niches, such as perinecrosis niche andperivascular niche.
: Our results suggest the potential biological & clinical significance of extrinsic ribosomal proteins in
GSCs development.

Copyright © The Japanese Pharmacological Society4-B-O12-3

一般演題（口頭）

Gene regulation during early and late phase of hypoxic response in cancer cells〇中山 恒
旭川医科大・医・薬理学

Cells in our body are often exposed to hypoxic environment. In such environment, cells regulate respiration andmetabolism for adaptation (hypoxic response). Hypoxic response not only plays a key role in maintaining
homeostasis, but also in different diseases, such as cancer. Hypoxia-Inducible Factor (HIF) is a central
transcription factor in hypoxic response. HIF is promptly up-regulated in hypoxia, and forms adapted cellularstate. HIF has multiple target genes, and they coordinately regulate hypoxic response. Recently, HIF specificinhibitor was developed, and it is currently under clinical trial. Alternatively, we have demonstrated that HIF is
downregulated, and CREB and NF-kB become activated during chronic phase of hypoxia. However, it remains
unclear how the expression pattern of hypoxia-responsive genes changes depending on the timing.
HIF becomes activated by a heterodimer formation of a and b subunits. There are three a subunit isotypes, andthey commonly bind to b subunit, ARNT. We have established ARNT knockout (KO) colon cancer HCT116
cells. These cells showed a clear inhibition of typical HIF target gene

. In contrast,, which isinduced under chronic hypoxia, was equally up-regulated in both wild type and KO cells, but its expression levelis significantly reduced in KO cells. Altogether, these results indicate that induction of hypoxic genes isdependent on transcription factors which are activated during chronic phase, however, basal expression of themis mediated by a HIF-dependent machinery.

Copyright © The Japanese Pharmacological Society4-B-O12-4

一般演題（口頭）

Inhibition of the mitochondrial shaping protein OPA1 restores lungadenocarcinoma cells sensitivity to gefitinib.

〇野口 雅史1,2、河野 晋3、ペラティエロ アンナ2、町田 雪乃4、柴田 圭太郎2、新谷 紀人1,5、河野 隆志6、後藤 典子3,7、高橋 智聡3,7、平尾 敦3,8、スコラーノ ルカ2、笠原 敦子3,7,8
1

和歌山県立医科大・薬・薬品作用学研究室、2パドヴァ大学・生物学部門・ベネト分子医学研究所、3金沢大・がん進展制御研究所、4日本獣医生命科学大・獣医病理学研究室、5大阪大・院薬・神経薬理、6国立がん研、7金沢大・新学術創成研究機構、8金沢大・ナノ生命科学研究所

Despite notable advances in chemotherapy protocols and targeted therapies, ensuing drug resistance limits theefficacy of cancer treatments, calling for the identifying druggable targets that can overcome chemo-resistance.
Here we show that the mitochondria-shaping protein OPA1 takes part in the resistance against the tyrosine
kinase inhibitor gefitinib in lung adenocarcinoma cells. In gefitinib-resistant lung cancer cells, OPA1 levels wereincreased, mitochondrial cristae structures were narrower and mitochondrial respiration increased. Genetic andpharmacological OPA1 inhibition in the resistant lung cancer cells sensitized them to gefitinib-induced
cytochrome c release and apoptosis.

, orthotopic tumors formed by the injection of gefitinib-resistantlung cancer cells were insensitive to gefitinib treatment, but a combination of gefitinib and OPA1 inhibitor
reduced tumor size and increased apoptosis. Our data identify the mitochondrial protein OPA1 as a downstream
factor that sustains gefitinib resistance and can be targeted to overcome chemo-resistance.

Copyright © The Japanese Pharmacological Society4-B-O12-5

一般演題（口頭）

Antitumor Activities by a Defucosylated Mouse–Dog Chimeric Anti-EGFR
Antibody in Canine Tumor Xenograft Models〇鈴木 裕之1、李 冠傑1、浅野 禎三2、田中 智大2、金子 美華2、加藤 幸成1,2
東北大・院医・分子薬理学分野、2東北大・院医・抗体創薬

1

The epidermal growth factor receptor (EGFR) contributes to tumor malignancy via gene amplification andprotein overexpression. Previously, we developed an anti-human EGFR (hEGFR) monoclonal antibody, EMab
-134, which detects hEGFR and dog EGFR (dEGFR) with high sensitivity and specificity by flow cytometry,western blotting and immunohistochemistry. In this study, we produced a defucosylated mouse‒dog chimericanti-EGFR monoclonal antibody, E134Bf. Kinetic analysis of the interactions of E134Bf with the canine
osteosarcoma cell line (D-17) and canine fibroblastic cell line (A-72) cells was conducted by flow cytometry.
The

D

for the interaction of E134Bf with the D-17 and A-72 cells was 5.5 × 10−10 M and 6.0 × 10−10 M,

respectively, indicating that E134Bf exhibits high affinity for D-17 and A-72 cells. Furthermore, E134Bf highly
exerted antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity against D-17 and A-72 cells.

administration of E134Bf significantly suppressed the development of D-17 and A-72

compared with the control dog IgG in mouse xenografts. These results indicate that E134Bf exerts antitumor
effects against dEGFR-expressing canine cancers and could be valuable as part of an antibody treatment
regimen for dogs.

Copyright © The Japanese Pharmacological Society4-B-O12-6

一般演題（口頭）

Effect of hyaluronic acid nanogels on the drug delivery of cyclosporine
administered subcutaneously in rats

橋本 桂樹1、宇佐美 歩樹1、森 理人1、伊藤 宏輔1、薄田 菜々子1、曽田 翠1、勝又 徹1、薮内 昂平2、
中井 貴士2、〇北市 清幸11

岐阜薬科大・薬・薬物動態学、2旭化成株式会社

Hyaluronic acid (HA) nanogel is derived by the partial modification of hyaluronic acid with cholesterol, causing
the formation of self-assembled nanometer-scale hydrogel in water. Thus, HA nanogels might contribute to alter
drug delivery of poorly water-soluble drugs. In this study, we tried to investigate the effect of HA nanogels byusing cyclosporine (CyA) in rats. HA nanogels (Asahi Kasei Co, 5 mL/kg) containing CyA (2.5-15 mg/kg) were
subcutaneously administered in male SD rats (Charles River) and whole blood samples were collected at thedesignated time-points up to 28 days. The concentrations of CyA were measured using LC-MS/MS 6045
(Shimadzu). In rats administered CyA without HA nanogel (control group), the concentration of CyA was
below the detection limit (3 ng/mL) on 7 days after administration. Some formulas of HA nanogel showed thesustained-release properties of CyA. Interestingly, one formula could be detectable up to at least 10 days afteradministration. HA nanogels would be useful as a tool to alter drug delivery of poorly water-soluble drugs suchas CyA.

Copyright © The Japanese Pharmacological Society4-B-O13-1

一般演題（口頭）
2+

Establishment of reconstituted depolarization-induced Ca release platform for
drug discovery of skeletal muscle diseases〇村山 尚1、呉林 なごみ1、冨田（沼賀） 拓郎2、小林 琢也1、中田 勉2、石田 良典3、森 修一3、影近 弘之3、山田 充彦2、
櫻井 隆1
順天堂大・医・薬理、2信州大・医・分子薬理、3医科歯科大・生材研1

In skeletal muscle, depolarization of the plasma membrane triggers Ca2+ release from the sarcoplasmic reticulum(SR),referred to as depolarization-induced Ca2+ release (DICR). DICR occurs via the type 1 ryanodine receptor(RyR1), which physically interacts with the dihydropyridine receptor Cav1.1 subunit in specific machinery
formed with additional essential components including β1a, Stac3 adaptor protein and junctophilins. It has
recently become clear that mutations in these components cause various skeletal muscle diseases. However, no
specific treatment has been developed yet. In this study, we established a high-throughput platform of thereconstituted DICR in HEK293 cells. The essential components were effectively transduced using baculovirus
vectors, and Ca2+ release was quantitatively measured with R-CEPIA1er, a fluorescent ER Ca2+ indicator. High[K+] depolarization triggered rapid Ca2+ release, indicating successful reconstitution of DICR. We tested several
known drugs modulating DICR. Whereas RyR1 inhibitors, dantrolene and Cpd1, suppressed DICR, twitch
potentiators, e.g., perchlorate, accelerated DICR. These results well reproduced the findings with the muscle
fibers and the cultured myotubes. The reconstituted DICR platform will be highly useful for drug discovery forskeletal muscle diseases.

Copyright © The Japanese Pharmacological Society4-B-O13-2

一般演題（口頭）

Actomyosin structure in smooth muscle cell treated with arachidonic acid〇田中 秀幸1、片山 豪2
1

帝京大・医、2高崎健康福祉大・人間発達

We have been searching for mechanism to induce smooth muscle contraction that is not associated withphosphorylation of the regulatory light chain (RLC) of smooth muscle myosin. We report that arachidonic acid
(AraA) stimulates ATPase activity of unphosphorylated smooth muscle myosin with maximal stimulation(Rmax) of 6.84 + 0.51 relative to stimulation by the vehicle and with a harf-maximal effective concentration(EC50) of 50.3 + 4.2 μM. In presence of actin Rmax was 1.72 + 0.08 and EC50 was 26.3 + 2.3 μM. Our
experiments with eicosanoids consisting of the AraA cascade suggested that they neither stimulated nor
inhibited the activity. Under conditions that did not allow RLC to be phosphorylated. AraA stimulated
contraction of smooth muscle tissue and culture cells with an Rmax of 1.45 + 0.07 and EC50 of 27.0 + 4.4 μM.
In addition to the ATPase activities of the myosin, AraA stimulated those of heavy meromyosin, subfragument 1(S1), S1 from which the RLC was removed, and a recombinant heavy chain consisting of the myosin head. The
stimulatory effects of AraA on these preparations were about two fold. The site of AraA action was indicated tobe the step-releasing iorganic phosphate (Pi) from the reaction intermediate of the myosin-ADP-Pi complex.
The enhancement of Pi relese by AraA was supported by computer stimulation indicating that AraA docked in
the actin- binding cleft oh the myosin motor domain. The stimulatory effect of AraA was detectable with both
unphosphorylated myosin and the myosin which RLC was fully phosphorylated. The AraA effect on both myosinforms was suggested to cause excess contraction and such as vasospasm.

Copyright © The Japanese Pharmacological Society4-B-O13-3

一般演題（口頭）
2+

Regulation of voltage-induced Ca release machinery by cell-cell fusion inskeletal myogenesis〇冨田 太一郎、山口 君空、三上 義礼、大島 大輔、赤羽 悟美
東邦大・医・統合生理One of the most dynamic processes during skeletal myogenesis is cell-cell fusion, which promotes skeletal
myoblast to form a large multi-nucleated myofiber equipped with EC-coupling machinery. Recently discovered
transmembrane protein Myomixer (Mymx) executes cell-cell fusion, although the physiological significance of
Mymx-mediated fusion on myogenic cell differentiation remains largely unknown. The current study focuses on
the intracellular Ca2+ signaling mechanism to understand the physiological importance of Mymx mediated cellfusion in regulating voltage-induced Ca2+release in the developing skeletal muscle cells. C2C12 cells were used
as an in vitro myogenesis model of skeletal muscle cells. We investigated intracellular Ca2+ release upon electricfield stimulation applied to differentiated Mymx-KO cells and Mymx-rescued cells. We found that the efficiency
of Ca2+ response was dependent on the Mymx gene expression. Importantly, expression levels of MyoD andmyogenin were almost unaltered by the gene rescue of Mymx, suggesting that the Mymx-dependent Ca2+

response is regulated independently of these transcription factors. In conclusion, we found a novel regulatory

linkage between Mymx expression and the voltage-induced Ca2+ release essential for EC coupling.

Copyright © The Japanese Pharmacological Society4-B-O13-4

一般演題（口頭）

Systemic administration of lipopolysaccharide derived from Escherichia coli, but
not Porphyromonas gingivalis, inhibits novelty-induced hyperlocomotion in mice

〇斉藤 幸治1、青野 悠里1、泉福 英信2、三枝 禎1
1

日本大・松戸歯・薬理、2日本大・松戸歯・感染免疫

Lipopolysaccharide (LPS), a component of the Gram-negative bacterial cell wall, activates Toll-like receptors(TLRs).

(Pg) appears to play a role in the development of periodontal disease.

studies have suggested that unlike LPS derived from

(Ec-LPS), which stimulates TLR4,

LPS derived from Pg (Pg-LPS) may inhibit the TLR4. Mice exposed to a novel environment show
hyperlocomotion that is inhibited by systemic administration of Ec-LPS. However, whether Pg-LPS influencesnovelty-induced locomotion is unknown. Therefore, we carried out an open field test to analyze the effects of
Pg-LPS. For comparison, effects of Ec-LPS were also studied. Male ddY mouse (25-30 g) were used. The
movement of each mouse in the open field was recorded for 30 min using a commercially available behaviouralanalysis system and the distance travelled (cm) was determined. Each compound was given intraperitoneally 4h
before the open field test. Ec-LPS 500 and 840 µg/kg, but not 100 µg/kg, inhibited novelty-induced increases in
distance travelled. Inhibition of hyperlocomotion by 840 µg/kg Ec-LPS was counteracted by co-administrationof the TLR4 antagonist TAK-242 (3.0 mg/kg). Pg-LPS (100, 500 or 840 µg/kg) failed to alter novelty-induced
locomotion. The present results provide

evidence that Ec- and Pg-LPS induce different effects. Thus,

Ec- but not Pg-LPS inhibits novelty-induced locomotor activity in mice by activating TLR4.

Copyright © The Japanese Pharmacological Society4-B-O13-5

一般演題（口頭）

DIF-1 promotes glucose uptake in mouse C2C12 myotube cells〇福永 優子1、中西 晃太郎1、杉田 光1、久保原 禅2
1

千葉科学大・危機管理学部・動物危機管理学科、2順天堂大・院スポーツ健康科学研究科・健康生命科学研究室

Differentiation-inducing factor-1 (DIF-1) isolated from the cellular slime moldhasbeen shown to promote glucose uptake in mouse 3T3-L1 fibroblasts and differentiated 3T3-L1 adipocytes (1).
DIF-1 promotes glucose uptake in the cells, at least in part, via an AMP kinase-dependent pathway (a PI-3
kinase/Akt-independent pathway), which is different from the insulin-induced glucose uptake pathway. In thisstudy, we investigated the actions of DIF-1 in skeletal muscle, the largest glucose-metabolizing tissue, using
C2C12 myotube cells.
During 15 h of incubation, DIF-1 at 1‒20 μM promoted glucose consumption (uptake) in a dose-dependent
manner in C2C12 myotube cells, while DIF-1 at 10‒20 μM was slightly toxic to the cells. DIF-1 (2 μM)induced glucose consumption was hardly inhibited with wortmannin (0.1 μM), a PI3K inhibitor, but waspartially inhibited with compound C (30 μM), an AMP kinase inhibitor. These results suggest that DIF-1
promotes glucose uptake in skeletal muscle cells via the same mechanisms as those in adipocytes and also thatDIF-1 may have therapeutic potential in the treatment of obesity and/or diabetes.
(1) Omata, W. et al. FEBS J. 274, 3392-3404. (2007).

Copyright © The Japanese Pharmacological Society4-B-O13-6

一般演題（口頭）

Assessment of learning achievement using self-assessment rubric in the roleplay for pharmacological education.

〇中村 正帆1、木村 武司2、長沼 史登1、吉川 雄朗3、岡村 信行1、柳田 俊彦41

東北医薬大・医・薬理、2京都大学医学部附属病院・総合臨床教育・研修センター、3東北大・院医・機能薬理、4宮崎大・

医看・臨床薬理Background. The role-play for pharmacological education (RPPE) provides a practical training focused on thebasis of drug therapy. Learning achievement in RPPE, however, has not been researched precisely. In this study,
we assessed the learning achievement in RPPE by using a self-assessment rubric.
Methods. Participants: Fourth grade medical students at Tohoku Medical and Pharmaceutical University who
took the online RPPE (oRPPE) in 2022. Measurements: Self-assessment rubric was presented to participants on
the first day of the course. Four following categories were scored on a five-point scale: I) Understanding: a.
health problems of the case in charge; b. drug therapy of the case in charge; c. role-play performed by theothers, II) Preparation, III) Performance and IV) Discussion and dialogue. Students were directed to assessthemselves during preparation and performance of oRPPE. Data collection and analysis: Scores were collected
after course completion. All data are presented as the mean ± standard error of the mean (S.E.M.).
Results. The self-assessment showed high scores in the following 3 categories: I) Understanding (a. 4.10 ±0.06; b. 4.10 ± 0.06; c. 3.94 ± 0.07), II) Preparation (4.32 ± 0.06) and III) Performance (4.21 ± 0.07). These
results indicate that RPPE could provide basic practice for drug therapy. It also implies that specification of
expected outcomes through rubric beforehand might be useful to guide self- and peer-learning during
preparation and performance. On the other hand, the score in IV) Discussion and dialogue (3.75 ± 0.09) mightreflect difficulties to discuss the whole process of clinical practice in each case, which requires advanced
professional competencies.
Conclusion. The self-assessment rubric is a useful tool for assessing achievement of learning in RPPE.

Copyright © The Japanese Pharmacological Society2-B-SS01-2

学生セッション（優秀発表賞候補）

CaMKII inhibition prevents the Dox-induced mitochondrial dysfunction without
the involvement of Drp1 or MCU〇プリヨノ アグン、三明 淳一朗、市原 克則、澤野 達哉、長田 佳子、今村 武史
鳥取大・医・薬理学・薬物療法学: Doxorubicin (Dox), an anticancer drug, is known to induce cardiac toxicity by causing
mitochondrial dysfunction. Although CaMKII and its phosphorylation targets, Drp1 to control mitochondrial
fission and MCU to control mitochondrial Ca2+ uptake, regulate mitochondrial homeostasis, the involvement ofthese molecules in the Dox-induced mitochondrial dysfunction remains unclear.
: To study the effects of Dox on mitochondrial homeostasis, we evaluated mitochondrial membranepotential (MMP), mitophagy, and mitochondrial Ca2+ content ([Ca2+]m) in H9C2 cells with the following
fluorescent dyes, JC-1, Mtphagy, and Rhod2-AM, respectively. To examine the activating effect of Dox on
CaMKII, we evaluated the phosphorylation levels of CaMKII by western blotting. To test the involvement of
CaMKII, Drp1, and MCU in the Dox-induced mitochondrial dysfunction, the specific inhibitors, KN-93, Mdivi-1, and Ru360, respectively, were used.
: Dox treatment dose-dependently reduced MMP and increased the number of cells with mitophagy and2[Ca +]m (p<0.05 in all). Dox treatment significantly increased the phosphorylation levels of CaMKII (p<0.05).
The inhibition of CaMKII suppressed the effects of Dox on the MMP and the mitophagy (p<0.05), but not on[Ca2+]m. Contrarily, the inhibition of Drp1 and MCU failed to suppress the decrease in MMP by Dox.
Similarly, the inhibition of Drp1 did not reverse the increase in mitophagy by Dox, nor did the inhibition of
MCU suppress the elevation of [Ca2+]m by Dox.
: These results suggested that activated CaMKII, but not Drp1 and MCU, is involved in the
impairment of MMP leading to Dox-induced mitochondrial dysfunction and that the excessive fission by Drp1
and the increased uptake [Ca2+]m by MCU are not the mechanism for the Dox-induced MMP reduction.

Copyright © The Japanese Pharmacological Society2-B-SS01-3

学生セッション（優秀発表賞候補）

Bioimaging analysis of a mouse model of atherosclerosis using radioisotopelabeled oxidized LDL as a probe targeting on foamy macrophages〇井澤 満1、河嶋 秀和2、奥野 結衣1、中谷 純菜1、武田 麻友子1、石原 慶一3、秋葉 聡3、高田 和幸1
1

京都薬大・統合薬科学、2京都薬大・放射性同位元素研セ、3京都薬大・病態生化学

Atherosclerotic plaques are formed by the accumulation of foamy macrophages, which phagocytose oxidized
low-density lipoprotein (LDL) in the intima of blood vessels. Although bioimaging to detect the accumulationof foamy macrophages is thought to be an effective tool for the prevention and treatment of atherosclerosis, it
has not yet been established. In this study, we examined the bioimaging of atherosclerotic plaques using humanoxidized LDL (oxLDL) labeled with 125I in apolipoprotein E knockout (ApoE-KO) mice, a mouse model ofatherosclerosis. Mouse bone marrow-derived macrophages markedly phagocytosed oxLDL but not intact LDL(LDL). The Oil Red O staining revealed that massive amounts of atherosclerosis plaques were formed in theaortic arch and aortic valve in ApoE-KO mice. In the study of single photon emission computed tomography(SPECT), distinct signals were detected in the aorta of ApoE-KO micetreated with 125I-labeled oxLDL but notwith 125I-labeled LDL. The local distribution of radioactivity was also detected by autoradiography. We further
confirmed the uptake of DiI-labeled oxLDL by macrophages accumulated in atherosclerotic plaques in ApoEKO mice. A possibility is suggested that bioimaging for the diagnosis of atherosclerosis could be developed withthe strategies such as application of radioisotope-labeled oxLDL.

Copyright © The Japanese Pharmacological Society2-B-SS01-4

学生セッション（優秀発表賞候補）

Protective effect of nobiletin against doxorubicin-induced cardiotoxicity inhuman iPS cell-derived cardiomyocytes.

〇岩下 賢士郎1、藤井 萌功1、稲井 誠2、坂本 多穂1、黒川 洵子11

静岡県立大・薬・生体情報分子解析学分野、2静岡県立大・薬・医薬品製造化学分野

Human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs) are increasingly being used for
cardiac safety evaluation, disease modeling, and regenerative medicine. To date, the majority of cardiotoxicity
studies have examined the acute drug effects. However, these studies lack information on the chronic effectsof cardiotoxic compounds. Here, motion vector prediction (MVP) method was employed to invasively quantify
contractile function over 10 days, then to test whether nobiletin has a protective effect against thecardiotoxicityof 8-days exposure to doxorubicin and erlotinib. The MVP method showed that doxorubicin (0.1 0.3 μM) significantly reduced contractility compared to erlotinib (0.3 - 3 μM), which has no cardiotoxicity,when administered for more than 5 days, and that the simultaneous addition of 0.03 μM nobiletin significantly
reduced this cardiac depression by doxorubicin (0.1 - 0.3 μM). These results suggest that nobiletin is
protective against the cardiotoxic effect of doxorubicin, and are promising for future drug discovery
applications.

Copyright © The Japanese Pharmacological Society2-B-SS01-5

学生セッション（優秀発表賞候補）

Neurofunctional phospholipids for inhibition of α-synuclein aggregation: A novel
therapeutic target for α-synucleinopathies〇唐木 達哉1、羽二生 久夫2、松田 佳和3、塚原 完1
1

長崎大・院医歯薬・創薬薬理学分野、2信州大・医・バイオメディカル研究所、3日本薬科大・薬・臨床薬理

Accumulation and aggregation of α-synuclein (α-Syn) are hallmarks of α-synucleinopathies such as dementia
with Lewy bodies. Therefore, aggregation of α-Syn is considered a priority target for drug development, andaggregation inhibitors are expected to reduce α-Syn toxicity and serve as therapeutic agents. Here, we reportthat certain lysophospholipids (LPLs) species behave as inhibitors for α-Syn aggregation. The LPLs are small
bioactive lipid molecules characterized by having a single carbon chain and a polar head group. The LPLs weused were extracted from Porcine Liver Decomposition Product (PLDP) which was previously reported toenhance cognitive function in healthy older adults. We found that the LPLs extracted from PLDP (PEL)
reduced α-Syn aggregation in cellular model. Especially, four species of LPLs contained in PEL strongly inhibit
α-Syn aggregation. Furthermore, we revealed that PEL increased normal cell viability in SK-N-SH cells. Finally
we approached the mechanism of the LPLsʻ inhibitory effect forα-Syn aggregation using

assay andevaluated influences to various cellular functions known to be disordered in lesion. Taken all together, thesestudies indicate that the LPLs would be beneficial as a possible therapeutic target in the treatment of αsynucleinopathies.

Copyright © The Japanese Pharmacological Society2-B-SS01-6

学生セッション（優秀発表賞候補）

Modulation of astrocyte activation by sphingomyelin〇門脇 凌、本田 拓哉、中村 浩之
千葉大・院薬・薬効薬理学

Astrocytes constitute about 20-40% of glial cells, making up the central nervous system (CNS). In the CNS,
astrocytes supply neurons with nutritional factors and maintain homeostasis of the extracellular environment
through the uptake and efflux of neurotransmitters. In neurodegenerative diseases, astrocytes change theirmorphology to the activated state and release inflammatory cytokines, which induce CNS inflammation andaggravate neurodegenerative diseases. Sphingolipids, one of the lipids, have been reported as a molecule
associated with astrocyte activation, but their involvement remains unclear. In this study, we evaluated astrocyte
activity by changing the levels of sphingomyelin (SM), one of the most abundant sphingolipids, using humanastrocyte/conditionally immortalized clone 35 (HASTR/ci35). Reduction of SM levels by knockdown of
sphingomyelin synthase 1 and/or 2 in HASTR/ci35 attenuated HASTR/ci35 activation by treatment ofinflammatory cytokines. Furthermore, selectively reducing SM levels by knockdown of ceramide transport
protein also attenuated the activation of HASTR/ci35. On the other hand, increasing the levels of SM byinhibition of neutral sphingomyelinase or addition of SM exogenously promoted activation of HASTR/ci35 bytreatment of inflammatory cytokines. These results suggest that SM positively regulates astrocyte activation.
Thus, regulating SM levels may provide a therapeutic target for astrocyte-induced CNS inflammation and a new
approach to treating neurodegenerative diseases.

Copyright © The Japanese Pharmacological Society2-B-SS01-7

学生セッション（優秀発表賞候補）

Neuroprotection and detection of Aβ by a low molecular weight compound
derived from natural product〇福田 愛菜1、中嶋 聡一2,6、尾田 好美2,6、西村 周泰1、花木 葵1、河嶋 秀和3、木村 寛之4、中村 誠宏2、松本 崇宏5、高田 和幸1
京都薬科大・薬・統合薬科学、2京都薬科大・薬・生薬学、3京都薬科大・薬・放射性同位元素研セ、4京都薬科大・薬・代1

謝分析学、5京都薬科大・薬・公衆衛生学、6NPR医薬資源研

In Alzheimerʻs disease (AD), acetylcholinergic (ACh) neurons are impaired at early pathological stage of AD,
and amyloid β (Aβ) oligomers are thought to be a crucial molecule for triggering neurodegenerative processes
in AD. In the present study, we first established anAβ oligomer-induced neuronal cell death modelusing human induced pluripotent stem cell (iPSC)-derived ACh neurons and-acyl isopeptide of Aβ, whichreverts to natural form of Aβ under the neutral pH conditions. In the processes of neurodegeneration in thismodel, Aβ was tightly attached dendrites and mitochondrial dysfunction was induced. We next identified an
Aβ-binding low molecular weight compound, plantainoside B, from the herbal extract of,which is used for memory enhancement in Ayurvedic medicine. Plantainoside B attenuated mitochondrial
dysfunction and exert neuroprotection against Aβ neurotoxicity. Moreover, the 125I-labeled plantainoside B
showed a high affinity to brain sections obtained from a model mouse of Aβ plaque formation and Aβoligomers in gel-loading experiments. Results indicate a possibility for the development of neurotheranostics
approach for the strategy of AD treatment.

Copyright © The Japanese Pharmacological Society2-B-SS02-1

学生セッション（優秀発表賞候補）

Impaired dendritic development is a common phenotype observed in primarycultured Purkinje cells expressing various SCA-causing proteins.

〇植田 恵梨香1、今野 歩2、平井 宏和2、倉内 祐樹1、香月 博志1、関 貴弘1,3
熊本大・院生命・薬物活性、2群馬大・院医・脳神経再生、3姫路獨協大・薬・薬理1

Spinocerebellar ataxia (SCA) is a group of autosomal-dominantly inherited ataxia and classified into SCA1-49
by the difference of causal genes, whose mutations include polyglutamine (polyQ)-expanded and missense
mutations. Purkinje cells (PCs) are neurons with highly developed dendrites and important for cerebellar
functions. We have previously revealed that impaired dendritic development is observed in primary cultured
PCs expressing missense mutant SCA14- and SCA21-causing proteins. We assume that various SCA-causing
proteins commonly impairs dendritic development in cultured PCs. In the present study, we expressed polyQexpanded (SCA1, 3 and 6) and missense (SCA34, 38, 41) mutant SCA-causing proteins in primary cultured PCs
and evaluated their dendritic development. Cerebellar primary cultures were prepared from E16 embryos of
Wistar rats and cultured for 3 weeks. SCA-causing proteins were expressed using adeno-associated viral vector.
Compared with wild-type proteins, all 6 SCA-causing proteins, including polyQ-expanded and missense
mutants, impaired dendritic development of primary cultured PCs. These findings indicates that impaired
dendritic development of cultured PCs is induced by various SCA-causing proteins and would be a common invitro phenotype of SCA and available for the exploration of novel SCA therapeutics.

Copyright © The Japanese Pharmacological Society2-B-SS02-2

学生セッション（優秀発表賞候補）

Involvement of PRMT5 in the activation of hepatic stellate cells〇山下 日菜子1、山口 桃生1、刀坂 泰史2、大岡 央1、森本 達也2、石川 智久11

静岡県立大・薬・薬理学、2静岡県立大・薬・分子病態

Liver fibrosis is a significant consequence of chronic liver diseases, where excess deposition of extracellularmatrix is caused by the activation of hepatic stellate cells (HSCs). The suppression of HSC activation istherefore regarded as a therapeutic target of liver fibrosis. The present study investigated the involvement ofprotein arginine methyltransferase 5 (PRMT5), which mediates genome organization and cell cycle regulation,in HSC activation. LX-2 cells, a human HSC cell line, were treated with TGF- β1 for 48 h in the presence of
PRMT5 inhibitors (EPZ015666 and JNJ64619178). The expression of α-smooth muscle actin (α-SMA) andtype I collagen α1 (COL1A1), activated HSC markers, were markedly increased by the TGF-β1 treatment.
PRMT5 inhibitors suppressed the increased expression of α-SMA and COL1A1 in a concentration-dependent
manner. Knockdown of PRMT5 also suppressed the TGF-β1-induced COL1A1 expression in LX-2 cells. RNAsequencing analysis showed that GO terms related to ECM production and SMAD signaling were enriched withRNA of LX-2 cells treated with the PRMT5 inhibitor JNJ64619178. These results suggest that PRMT5
promotes HSC activation, possibly depending on the SMAD signaling pathway, and therefore might be a target
for the prevention and treatment of liver fibrosis.

Copyright © The Japanese Pharmacological Society2-B-SS02-3

学生セッション（優秀発表賞候補）

Establishment of assessment system for anti-fibrotic activity using an ex vivo
hepatic fibrosis model〇髙橋 亮汰、山口 桃生、森野 純鈴、岡部 磨幸、大岡 央、石川 智久
静岡県立大・薬・薬理

Currently, there is no effective treatment for liver fibrosis.

studies on the activation of hepatic stellate

cells (HSCs), which is responsible for liver fibrosis, have been used as a drug screening system for it. However,even if some drug shows an inhibitory effect on HSC activation, its anti-fibrotic effect is required to be
confirmed in liver fibrosis animal models, which further requires a lot of time and cost. In the present study, we
tried to establish anmodel of liver fibrosis using precision-cut liver slices (PCLSs) to solve suchproblems. PCLSs of 250 µm thickness were prepared from male C57BL/6J mice using a vibratome and cultured
in RPMI medium in a 5% CO2 incubator. Although cellular ATP content was decreased on day 1 compared to
day 0, it was then maintained until day 5, suggesting that the ex vivo model is viable for at least 5 days.
Treatment with Et-OH (50, 100 mM), one of the liver injury stimuli, for 5 days increased mRNA expression of
Acta2 and Col1a1, liver fibrosis markers, in PCLSs. DIF-1 (50, 100 µM), which has an anti-fibrotic effect,
significantly suppressed the Et-OH-induced increases in the markers. These results suggest that themodel using PCLSs is useful as a drug screening system for the development of drugs for treatment of liver
fibrosis.

Copyright © The Japanese Pharmacological Society2-B-SS02-4

学生セッション（優秀発表賞候補）

The development of biologics inhibiting IL-33 signaling on the basis of the signaltransduction mechanisms〇山本 里彩1、安藤 智暁1、貝谷 綾子1、伊沢 久未1、前原 明絵1、北浦 次郎11

順天堂大・院医・アトピー疾患研究センター、2順天堂大・医・5年生

IL-33 contributes to the pathogenesis of allergic diseases. Upon the binding of IL-33 to a membrane receptor
ST2, IL-1 receptor accessory protein (IL-1RAcP) is recruited to form a heterodimeric receptor complex whichtransmits a signal. On the other hand, a soluble form of ST2 acts as an endogenous inhibitor of IL-33
signaling. Here, we aimed to develop the biologics inhibiting IL-33 signaling. First, we generated IL-33 reporter
cell lines, in which IL-33 stimulation induced NFkB-driven expression of DsRed. We then generated IL-33trapFc which is composed of extracellular domains of IL-1RAcP and ST2 fused to the Fc portion of human IgG.
Notably, IL-33trap-Fc more strongly suppressed IL-33-stimulated DsRed expression in the reporter cells thanST2-Fc which is composed of an extracellular domain of ST2 fused to human IgG Fc. Consistently, IL-33trap-Fc
remarkably inhibited IL-33-stimulated IL-6 production of bone marrow-derived mast cells as compared with
ST2-Fc. Moreover, intraperitoneal administration of IL-33trap-Fc efficiently inhibited IL-33-stimulated
eosinophil accumulation in vivo. Taken together, these results indicated that IL-33trap-Fc was effective ininhibiting IL-33 signaling. We also attempt to generate the biologics that bind to ST2 but fail to recruit IL-1RAcP, thereby blocking IL-33 signaling.

Copyright © The Japanese Pharmacological Society2-B-SS02-5

学生セッション（優秀発表賞候補）

L-Carnitine supplementation attenuates lenvatinib-induced muscle impairment
without diminishing its anti-angiogenesis efficacy〇靖 崢1、射場 智大2、内藤 尚道2、許 平平1、盛重 純一1、長田 直人1、大久保 裕直3、安藤 仁1
金沢大・院医薬保健・細胞分子機能学、2金沢大・院医薬保健・血管分子生理学、3順天堂大学医学部附属練馬病院・

1

消化器内科

Lenvatinib (LEN), an oral tyrosine kinase inhibitor, is widely used to treat several types of advanced
cancers but often causes muscular adverse reactions. Our previous study revealed that LEN reduces L-carnitine(L-CAR) content, expression of carnitine-related genes, and mitochondrial function in the skeletal muscle.
Therefore, the present study aimed to investigate whether L-CAR supplementation prevents LEN-induced
muscle impairment without affecting its anti-angiogenesis effect.
Eight-week-old male Wistar rats were divided into four groups and administrated orally once dailyfor 2 weeks with vehicle, LEN (2 mg/kg/day), LEN + L-CAR (150 mg/kg/day), or LEN + L-CAR (300 mg/kg).
In the in vitro studies, differentiated C2C12 myocytes, HUVECs, and mouse aorta were treated with LEN (0.1
and/or 1 μM) and L-CAR (1.6, 6.4, and 25.6 mM).
L-CAR supplementation significantly attenuated LEN-induced deleterious effects on L-CAR content,
expression of carnitine-related (OCTN2, CPT1, CACT, and CPT2) and OXPHOS genes in the skeletal muscleof rats. In addition, L-CAR prevented LEN-induced reductions in mitochondrial function (ATP content andmembrane potential) in C2C12 myocytes. Furthermore, L-CAR did not affect the anti-angiogenesis action of
LEN assessed by the tube formation and ring assays.
These results suggest that L-CAR supplementation can alleviate the adverse reactions of LEN in
the skeletal muscle without reducing its antineoplastic effect.

Copyright © The Japanese Pharmacological Society2-B-SS02-6

学生セッション（優秀発表賞候補）

Development of fast-dissociating recombinant antibody probes for multiplexed
super-resolution molecular mapping〇張 千里1、宮本 章歳1、有森 貴夫2、高木 淳一2、渡邊 直樹1,3
京都大・院生命科学・分子動態生理学、2大阪大・蛋白質研究所・分子創製学研究室、3京都大・院医・神経薬理学

1

We report a mutagenesis strategy that can effectively increase the dissociation rate of antibodies by orders ofmagnitude without compromising the binding specificity. Single-molecule localization microscopy greatly
surpasses the diffraction limit of conventional optical microscopy. The imaging fidelity and labelling density,
however, are limited by spatial interference between bulky antibodies in a confined resolved area. IRIS has
overcome the problem using exchangeable probes that transiently bind to endogenous targets. In our previous
research, generation of fast-dissociating IRIS probes has been challenging. In the present study, we havedeveloped a new mutagenesis strategy that make it feasible to generate IRIS probes from the repository of offthe-shelf antibodies. We successfully generated dozens of IRIS probes and demonstrate multiplexed localizationof endogenous proteins in primary neurons that visualizes small synaptic connections. Our fast-dissociatingprobes achieved 4-fold higher label density than conventional super-resolution approaches. Thus, IRIS could
visualize the feature of synaptic components with higher fidelity. In addition, the mutagenesis strategy will
provide more applications for high affinity antibodies developed in pharmaceutical research, such as superresolution imaging based disease diagnosis and biomarker identification.

Copyright © The Japanese Pharmacological Society1-B-SS03-1

学生セッション

Effects of 2-carba- cyclic phosphatidic acid derivatives on IL-1β-stimulated
human chondrocytes〇吉井 雅起1、諸星 俊郎2、塚原 完1
1

長崎大・院医歯薬・創薬薬理学分野、2SANSHO(株)Osteoarthritis (OA) is a common joint disease characterized by the breakdown of subchondral bone andcartilage damage, most often affecting middle-aged and elderly people. Although the etiology of OA is still
unknown, some reports suggest that inflammatory factors such as interleukin (IL)-1β mediate the progression
of OA. In order to investigate the effect of IL-1β and the possibility of treatment for OA, we used 2-carba-cyclic
phosphatidic acid (2ccPA) and its derivatives on human chondrocytes. 2ccPA is a synthesized phospholipid
based from a bioactive phospholipid mediator: cyclic phosphatidic acid (cPA). It is previously reported that
2ccPA exhibits anti-inflammatory and chondroprotective effects on an OA animal model. 2ccPA and its ringopened body (ROB) derivative significantly suppressed IL-1β-induced upregulation of IL-6, matrix
metalloproteinase-13, and cyclooxygenase-2, as well as the degradation of type II collagen and aggrecan.
However, the other two derivatives, the deacylated body and the ring-opened deacylated body showed little
effect on IL-1β-exposed human chondrosarcoma cell-line. These data suggest that acyl chain of 2ccPA and
ROB is essential for anti-inflammatory effect on OA. Taken together, this study provides evidence that 2ccPAand ROB would be a novel therapeutic agent for OA.

Copyright © The Japanese Pharmacological Society1-B-SS03-2

学生セッションCan Cannabidiol affect the peripheral circadian clock in PER2::LUC mice?

〇WANG HSIAO HSIEH3、野崎 千尋1、柴田 重信31

早稲田大・国際理工学センター・Major in Bioscience、2University of Bonn・Institute of Molecular Psychiatry、3早稲田

大・理工・電気．情報生命工学科

Cannabidiol (CBD) is the second major cannabinoid which is often said to improve anxiety and sleep with nopsychoactive effect. It is said that CBD can promote the production of wake-related neurotransmitters such as
dopamine. Further, co-injection of CBD and THC increased sleeping time in rats. However, no study has been
done for the in-vivo effect of CBD to circadian clock. Present study aimed to elucidate whether CBD can modify
peripheral circadian rhythm.
PER2::LUC knock-in female mice were used to determine the effect of CBD to the peripheral clock. Mice weredivided into six groups: CBD isolate (99% crystalline) in MCT oil, CBD isolate in 5% EtOH/5%
cremophor/water, and water-soluble CBD nanopowder in water, with respective vehicle controls. Each drug wasorally administered at ZT4 or ZT16 for three days, then PER2 gene expression in the liver, kidney, andsubmandibular gland is observed by in-vivo imaging.
We found that phase advance in the liver and the submandibular gland only happens when MCT or CBD in
MCT (CBD/MCT) was administered at ZT4. However, the phase advance did not differ between the MCT and
CBD/MCT groups. Furthermore, neither the vehicle nor the CBD affected the peripheral clock whenadministered at ZT16.
This study suggested that not CBD but rather MCT oil affect the circadian clock in mice. As MCT oil iscommonly used as a base for CBD products, we propose that MCT might be the possible factor that affects thecircadian clock and cause rhythm improvement. Therefore further detailed studies on the effect of CBD
products will be needed.

Copyright © The Japanese Pharmacological Society1-B-SS03-3

学生セッション

Derivatized-imaging mass spectrometer revealed the effect of theanine formonoamine metabolism〇武藤 誠1、鹿野 仁美1、安部 綾2、小関 誠2、平 修1
福島大学・農学群・食農学類、2太陽化学株式会社・ニュートリション事業部

1

L-Theanine (LTE) is a derivative of glutamic acid, which is abundant in tea leaves and contributes to the umami
and sweetness of tea. In recent years, its effects on the brain, such as relaxation, have been attracting attention.
In this study, we used imaging mass spectrometry (IMS) to visually analyze the changes in the catecholamine
system in the brain after the administration of LTE. In IMS, we applied derivatization reagent to improvedetection limit. Simultaneous imaging of catecholamines, LTE and g-aminobutyric acid (GABA) is particularly
useful to understand a metabolic pathway. We investigated whether symptom of depression is improved or not
by free drinking of LTE water. The mice that are freely drinking of theanine (Group 1), symptoms of depression
was milder than that of drinking of water (Group 2). IMS showed dopamine (DA) marginally produced from
caudate putamen from Group 2, but DA was produced from the Group 1. Interestingly, GABA increased at
hypothalamus nucleus paraventricularis (PVN), which controls eating amount, from Group 1 compared withGroup 2. We hypothesized an increasing of GABA at PVN works an appetite stimulation. Practically, anappetite of mouse in Group 1 was not decreased compared with that in Group 2. IMS visually gives us thepathway of catechol amine and improvement of depression by theanine.

Copyright © The Japanese Pharmacological Society1-B-SS03-4

学生セッション

The Hippo pathway kinase Lats1/2 inhibition increases slow- and fast-twitch
fibers via activation of Tead cofactors.

〇堀 直人、鈴木 孝行、荒木 拓郎、石川 千夏、南須原 杏律、高野 博之、山口 憲孝
千葉大・院薬・分子心血管薬理学研究室【Background】
Skeletal muscles are composed of slow-twitch fiber and fast-twitch fiber. Transcriptional factor Tead induces
muscle differentiation with its cofactor Taz. However, whether Tead controls slow- and fast-twitch fiber is
unknown. Thus, we focused on Vgll3, another cofactor of Tead, and investigated its role in muscle
differentiation.
【Result】
We established Vgll3 expressing C2C12 cells and differentiated these cells. Vgll3 expressing cells showed
upregulation of muscle differentiation markers and an increase in slow-twitch fiber marker Myh7 with adecrease in fast-twitch fiber marker Myh4. These results suggest that Vgll3 increases slow-twitch fiber.
Recent studies reported Taz activation induces Vgll3 expression. Thus, we used the Lats1/2 inhibitor, which
induces Taz activation. Lats1/2 inhibitor induces Vgll3 expression and upregulation of Myh7 and Myh4. This
result suggests that Lats1/2 inhibition induces Taz activation to increase fast-twitch fiber and Vgll3 expression
to increase slow-twitch fiber simultaneously.
【Conclusion】
Our results suggested that Vgll3 increases slow-twitch fiber, and Lats1/2 inhibition increase slow- and fasttwitch fiber via activation of Vgll3 and Taz. Aging significantly decreases fast-twitch fiber and causes older adults
injury. Thus, Tead and its cofactors are important for aging treatment.

Copyright © The Japanese Pharmacological Society1-B-SS03-5

学生セッション

Identification of novel effects of Eliglustat and investigation its affects against
idiopathic pulmonary fibrosis (IPF)〇久留宮 慧臣、本田 拓也、中村 浩之
千葉大・院薬・薬効薬理学【Background, Purpose】
Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, irreversible, and ultimately fatal lung disease. IPF
occurs due to TGF-β1/Smad signaling activation.
Previously, we observed that eliglustat, glucosylceramide synthase inhibitor exhibited anti-fibrotic effects against
nomal human lung fibroblast. So, we checked whether eliglustat exhibits anti-fibrotic effects also against IPF
patient-derived cells (IPF cells), and tried to elucidate its mechanism.
【Results, Discussion】
First, we tested the anti-fibrotic effects of eliglustat in IPF cells. Treatment of IPF cells with eliglustat similarly
suppressed the up-regulation of fibrotic proteins such as α-SMA and collagen by TGF-β1. Eliglustat had noeffects on phosphorylation and translocation to the nucleus of Smad. The knockdown of glucosylceramide
synthase did not inhibit the up-regulation of α-SMA by TGF-β1. These results suggest that eliglustat inhibits
fibrotic protein transcription by Smad independently of its inhibitory effect against glucosylceramide synthase.
Next, we focused on sterol regulatory element-binding protein2 (SREBP2) to elucidate the anti-fibrotic
mechanism of eliglustat. SREBP2 regulates intracellular cholesterol levels and is known to inhibit the transcript
activity of Smad. Treatment of IPF cells with eliglustat induced translocation of SREBP2 to the nucleus and upregulation of downstream genes of SREBP2. Inhibition of SREBP2 attenuated the eliglustat-induced downregulation of α-SMA expression. These results suggest that eliglustat exhibits anti-fibrotic effects throughactivation of SREBP2.

Copyright © The Japanese Pharmacological Society1-B-SS03-6

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Regulation of neuron-astrocyte communication via ATP/P2Y1 signaling by
microglia〇鈴木 秀明1,2、繁冨 英治1,2、平山 幸歩1、高橋 由香里3、池中 一裕4、田中 謙二5、加藤 総夫3、尾藤 晴彦6、小泉 修一
1,2
1

山梨大・院医・薬理、2山梨大・院医・山梨GLIAセンター、3慈恵医大・神経科学、4生理研・分子神経生理、5慶應大・医・

先端研・脳科学、6東京大・院医・神経生化学

P2Y1 receptor is upregulated in astrocytes in many neurological diseases. We have previously shown thatelevated P2Y1 receptor expression in astrocytes causes neuronal hyperexcitability by enhancing neuronastrocyte communication in the hippocampal CA1 region. However, contribution of microglia to the astrocytic
P2Y1 receptor-mediated neuron-astrocyte communication is not known, despite the fact that microglia are also
activated in such pathological conditions. To this end, we attempted to investigate the role of microglia in
astrocyte P2Y1 receptor signaling by depleting microglia with a CSF1 receptor antagonist, PLX5622. The
results are summarized in the following two points: 1) Microglia depletion increased

gene expression inastrocytes and enhanced Ca2+ signal via P2Y1 receptor, indicating that microglia would have a role to inhibit
P2Y1 receptor expression in astrocytes. 2) Microglia depletion prolonged the time required for degradation ofexogenously applied ATP. Because microglia highly express an ATP degrading enzyme CD39, they would play acentral role in shutting-off of P2Y1 receptor signals by metabolizing ATP. Taken together, it is suggested thatmicroglia would also play an important role in neuron-astrocyte communication via 2 different modes, i.e.,
inhibition of P2Y1 receptor expression and degradation of extracellular ATP.

Copyright © The Japanese Pharmacological Society1-B-SS04-1

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Mechanistic analysis for suppression of neuropathic pain in circadian clock genedeficient mice

〇山川 稚葉1、安河内 冴1、鶴留 優也4、牛島 健太郎4、鶴田 朗人1,2、松永 直也3、小柳 悟1,2、大戸 茂弘11

九州大・院薬・薬剤学、2九州大・院医薬・グローカルヘルスケア、3九州大・院薬・薬物動態学、4山口東京理科大・薬・薬剤学，製剤学

Because the expression of up to 10% of genes is under the control of the circadian machinery consisting of clock
genes, it should not come as a surprise that the disfunction of clock gene affects the onset and/or state of various
disease. Diurnal variations in pain hypersensitivity are common in chronic pain disorders, but pathologicalrelevance of clock genes in neuropathic pain hypersensitivity remains unknown. In this study, we investigated
the threshold of mechanical pain hypersensitivity in peripheral sciatic nerve-ligated (PSL) animals and foundthat clock gene deficient mice (observed in wild-type mice, PSL-mice) failed to develop the neuropathic pain hypersensitivity. As
mice also activation of glial cells in the dorsal horn of the spinal cord,

as well as increased expression of pain-related molecules. On the other hand, the descending pain suppressor
system and endocannabinoid system were upregulated inmice, suggesting that the suppression

mechanism against neuropathic pain is enhanced by dysfunction of clock gene. Therefore,

mice are lesslikely to develop pain hypersensitivity even when peripheral nerves are injured. These findings indicate that
endogenous pain suppression system are under the control of circadian clock. Identification of circadian clock
controlled pain suppressor molecule would be a therapeutic target for treatment of neuropathic pain.

Copyright © The Japanese Pharmacological Society1-B-SS04-2

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A subset of spinal dorsal horn inhibitory interneurons crucial for analgesic effectassociated with spinal noradrenaline〇末藤 大智1、石橋 忠幸1,2、吉川 優1、山浦 健2、津田 誠1
九州大・院薬・薬理学分野、2九州大・院医・外科学講座 麻酔・蘇生学

1

Pain information transmission/processing in the spinal dorsal horn (SDH) is strongly controlled by descending
neurons from the brain. One of the major neurotransmitters of descending pathways is noradrenaline (NA).
Descending NAergic neurons from the locus coeruleus (LC) is known to produce analgesic effects via activationof inhibitory interneurons in the SDH. However, the identity of the inhibitory interneuron subset in the SDH ispoorly understood. Recently, we have found a subset of the SDH inhibitory interneurons captured by adenoassociated viral (AAV) vectors incorporating a neuropeptide Y promoter (AAV-NpyP+) that is crucial forneuropathic allodynia. Here, we showed that this neuronal subset is a major target of spinal NA to inhibit pain
information transmission/processing. Whole-cell patch-clamp recordings using spinal cord slices revealed that
NA predominantly depolarizes AAV-NpyP+ neurons. This effect was suppressed by a pharmacological blockade
and genetic knockdown of α1B-adrenoceptor (AR) in AAV-NpyP+ neurons in the SDH. Furthermore, we foundthat the analgesic effect of duloxetine on neuropathic pain which is associated with an increase in the spinal NA
level by inhibiting NA reuptake into presynaptic terminals is reduced by AAV-NpyP+ neuron-selective
knockdown of α1B-ARs. These results indicate that α1B-ARs expressed in AAV-NpyP+ neurons would be atarget of spinal NA presumably from descending LC neurons and contribute to the analgesic effect of
duloxetine. Thus, spinal α1B-ARs could be a new therapeutic target.

Copyright © The Japanese Pharmacological Society1-B-SS04-3

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Paclitaxel-induced peripheral neuropathic pain formation contributes to synaptic
plasticity in the spinal dorsal horn.

〇山田 果琳、澤幡 雅人、久米 利明、歌 大介
富山大・院医薬・応用薬理学研究室

Paclitaxel (PTX) is a typical anticancer drug that induces peripheral neuropathy and significantly
reduces patients' quality of life. Previous our study showed gabapentinoids, such as mirogabalin, attenuate PTXinduced peripheral neuropathic pain by acting on the spinal dorsal horn. PTX-induced peripheral neuropathy
has so far focused on peripheral inflammation, but it is possible that changes in the spinal dorsal horn may
contribute to PTX-induced peripheral neuropathy. In this study, we investigated the mechanism of synaptic
plasticity in the spinal dorsal horn by using electrophysiological and immunohistochemical analysis.
We administered a single intraperitoneal dose of PTX 5 mg/kg to C57BL/6NCr mice. We analyzed
the frequency of spontaneous and von Frey filament (vFF; 0.69 mN) evoked firing in spinal dorsal horn neurons
by using

extracellular recording. Immunohistochemical staining was performed on spinal cord (L4-6)

slices.
Electrophsiological data showed the frequency of spontaneous and vFF evoked firing in spinal dorsal
horn neurons were significantly enhanced in PTX model mice.
The levels of the neuronal activation marker c-fos were increased with mechanical allodynia formation in PTXmodel mice. Glia-associated makers Iba1 and GFAP also showed chronological changes after PTX-treatment.
The present study suggests that synaptic plastic changes occur not only in the periphery but also inthe spinal dorsal horn in the PTX-induced peripheral neuropathic pain model.

Copyright © The Japanese Pharmacological Society1-B-SS04-4

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The effect of REV-ERB agonist on nociceptive hypersensitivity inmonoiodoacetate-induced osteoarthritis model〇橋爪 宥樹、元成 初寧、中村 庸輝、中島 一恵、森岡 徳光
広島大・院医系 (薬)・薬効解析

Osteoarthritis (OA) is characterized by pain caused by inflammation and degradation of cartilage matrix in
joint. Although the number of patients is expected to increase due to the aging of the population, the use ofanti-inflammatory drugs, which is the main therapeutics, may not be effective in pathology of OA. REV-ERBs
are one of nuclear receptors involved in a wide range of physiological functions. We have previously shown that
REV-ERBs were expressed in primary cultured chondrocytes, and REV-ERB agonist suppresses theupregulation of proinflammatory cytokines and matrix degradation enzymes in these cells under inflammatory
conditions. However, the role of REV-ERBs in pathogenesis of OA is not clear. Thus, we investigated the effectof REV-ERB agonist on nociceptive hypersensitivity in monoiodoacetate (MIA)-induced OA model. SR9009, a
REV-ERB agonist, was administered intraarticularly twice a week, starting 3 days after MIA administration.
Mechanical thresholds were measured by the von Frey test. MIA induced mechanical hypersensitivity from day
3 after administration, which persisted at least until day 28. Administration of SR9009 significantly ameliorated
mechanical hypersensitivity from day 14 after MIA administration. These results suggest that activation of REVERB might induce an analgesic effect on OA pain.

Copyright © The Japanese Pharmacological Society1-B-SS04-6

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A prototype of the microsensing system for in vivo drug monitoring in the skinwith diamond electrode〇Ahmad Norzahirah Binti1、澤村 晴志朗1、緒方 元気2、栄長 泰明2、日比野 浩1
1

大阪大・院医・統合薬理学、2慶應義塾大・理工・化学

Monitoring of plasma drug concentrations is required for effective pharmacotherapy. Repetitive collection ofwhole blood followed by analysis of plasma samples with conventional methods delays representation of crucial
results. Skin is an easily accessible organ; a portion of systemically circulating drug molecules is diffused to thedermal interstitial fluid. Thus, the compoundʻs pharmacokinetics (PK) in the fluid mirrors the plasma PK. To
approach such local dermal space, here we describe a microsensing system with a needle-type boron-doped
diamond (BDD) electrode, which detects chemical compounds by redox reaction. As a test analyte we chose an
anticancer drug, doxorubicin. In an in vitro experiment with a BDD microsensor, doxorubicin elicited a current
in response to applied negative potential. Calibration curve covered the therapeutic window (10−100 nM). The
sensorʻs performance was also tested in the collected interstitial fluids. Finally, the sensor was inserted into thedermis layer in anesthetized live rats; after doxorubicin was intravenously injected, the local PK was tracked for>1 hour with the Cmax and Tmax 3.1 ± 1.4 nM and 33.6 ± 20.6 mins, respectively (n = 7). By combining aformula linking the local measurements to plasma data, this microsensing system may be applicable to real-timemonitoring of systemic PK.

Copyright © The Japanese Pharmacological Society1-B-SS05-1

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Three-dimensional structural analysis of pharmacokinetics-related membraneprotein P-glycoprotein using cryo-electron microscopy〇濱口 紀江1,2、安達 成彦3、守屋 俊夫3、川崎 政人3、安田 賢司2、千田 俊哉3、小笠原 諭2、村田 武士2
千葉大・院医薬・薬理学、2千葉大・理・構造生物学、3高エネ機構・物構研・構造生物

1P-glycoprotein (P-gp) is mainly found in the cell membrane of the small intestine and blood-brain barrier invivo, and is responsible for the extracellular transport of cytotoxic hydrophobic compounds. P-gp is known totransport many pharmaceutical compounds as substrates. If we can understand the substrate recognition
mechanism of P-gp, it will be possible to design pharmaceutical compounds that are not recognized by P-gp.
Recently, the complex structures of human P-gp with substrates and inhibitors have been reported by singleparticle analysis using Cryo-EM, and the differences in the binding pockets of substrates and inhibitors havebeen clarified. However, a detailed understanding of how P-gp can identify compounds as substrates orinhibitors has not been achieved. In this study, we aim to elucidate the detailed substrate recognition
mechanism by elucidating and comparing multiple complex structures of P-gp and compounds. First, weestablished a system for expression and purification of human P-gp. Further, we have established a simple
system for reconstitution into Nanodisc. Recently, we succeeded to obtain the 3D structure at the highest
resolution (2.93 Å) as human P-gp. In this presentation, we will introduce the expression, purification, andNanodisc reconstruction systems of P-gp and the obtained 3D structures.

Copyright © The Japanese Pharmacological Society1-B-SS05-2

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Caveolin-1 regulates ATP signaling mediated by P2X7 receptor in proinflammatory macrophages.

〇澤井 優輝、鈴木 良明、近藤 るびい、今泉 祐治、山村 寿男
名古屋市大・院薬・細胞分子薬効解析[Background] Macrophage (Mφ) plays crucial roles in immunity and its dysfunction leads to the chronic
inflammatory diseases such as arteriosclerosis. Several Mφ functions are modulated by the activation ofionotropic purinergic P2X7 receptor. Caveolin-1 (Cav-1) enables effective intracellular Ca2+ signaling byaccumulating ion channels within caveolae domain. In this study, we analyzed the functional coupling between
Cav-1 and P2X7 receptor using Cav-1 knockout (Cav-1 KO) mice.
[Methods] In murine bone marrow-derived Mφ (BMDM), the expression of Cav-1 was analyzed by real-time
PCR and Western Blotting. Interaction of Cav-1 and P2X7 receptor was analyzed by proximal ligation assay. Ca2+

influx, K+ efflux and reactive oxygen species (ROS) production were measured with confocal microscopy. Cell

death was analyzed by LDH assay.
[Results] The expression of Cav-1 was increased by LPS (lipopolysaccharide)-induced inflammatory stimulationin BMDM. Cav-1 was interacted with P2X7 receptor. Thereafter, ATP-evoked Ca2+ influx and K+ efflux wereincreased in Cav-1 KO BMDM. ROS production and cell death evoked by ATP were also enhanced in Cav-1 KO
BMDM.
[Conclusion] Cav-1 suppresses the activation of P2X7 receptor and modulates immune responses in Mφ. This
study may lead to the development of novel drugs for chronic inflammatory diseases.

Copyright © The Japanese Pharmacological Society1-B-SS05-3

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Pore opening, not voltage sensor movement, underpins the voltage-dependenceof facilitation by a hERG blocker.

〇河野 諒太朗1、古谷 和春1,2、一藁 南1、足立 亮1、Clancy Colleen2、Sack Jon2、喜多 紗斗美11

徳島文理大・薬・薬学科、2カリフォルニア大学デービス校・医・生理学

A drug that blocks the cardiac myocyte voltage-gated K+ channels encoded by the hERG carries a potential riskof long QT syndrome and life-threatening cardiac arrhythmia. Interestingly, certain hERG blockers can also
facilitate hERG activation to increase hERG currents, which may reduce proarrhythmic potential. However, themolecular mechanism remains unclear. The hallmark feature of the facilitation effect by hERG blockers is that adepolarizing preconditioning pulse shifts voltage-dependence of hERG activation to more negative voltages.
Here we utilize a D540K hERG mutant to study the mechanism of the facilitation effect. D540K hERG is
activated by not only depolarization but also hyperpolarization. With D540K hERG, we find that nifekalant, ahERG blocker and Class III antiarrhythmic agent, blocks and facilitates not only current activation by
depolarization but also current activation by hyperpolarization, suggesting a shared gating process upon
depolarization and hyperpolarization. Moreover, in response to hyperpolarizing conditionings, nifekalantfacilitates D540K hERG currents but not wild-type currents. Our results indicate that induction of facilitation iscoupled to pore opening, not voltage per se. We propose that gated access to the hERG central cavity underlies
the voltage-dependence of induction of facilitation.

Copyright © The Japanese Pharmacological Society1-B-SS05-4

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Development of a novel drug targeting TRPC3/C6 channels〇白戸 真美子
京都大・大学院工学研究科・合成生物・化学専攻

A group of TRPCs, TRPC3, TRPC6, and TRPC7, form Ca2+-permeable channels directly activated bydiacylglycerol (DAG) and play important roles in regulating neuronal survival and dendritic growth,cardiovascular fibrosis

andthrough regulation of Ca2+signaling. Various compounds targetingthese TRPC channels have been developed for the treatment of serious diseases such as sudden pulmonary
fibrosis and chronic nephropathy. However, none of these compounds have yet reached clinical application, andtherefore development of new TRPC3/C6/C7 inhibitors has been much-needed. Here, we have developed apiperazine derivative targeting TRPC3/C6 channels. This compound suppressed receptor-activated Ca2+influx
in a dose-dependent manner in human embryonic kidney cells 293 expressed with human TRPC3 or TRPC6(TRPC3, IC50 = 0.086: TRPC6, IC50 = 0.034µM). This drug showed no significant inhibitory or stimulatory
effect on other TRPs including TRPC7. Interestingly, during isolation of human TRPC7, we obtained a new
splice variant of human TRPC7; we are in the process to characterize biophysical and pharmacologicalproperties of the variant that has a deletion in one of the functionally critical domains.

Copyright © The Japanese Pharmacological Society1-B-SS05-5

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2+-

TMEM16A-mediated Ca -activated Cl currents is increased in portal vein smoothmuscle cells from caveolin 1-deficient mice

〇川田 成紀、近藤 るびい、鈴木 良明、山村 寿男
名古屋市大・院薬・細胞分子薬効解析

In vascular smooth muscles, the activity of Ca2+-activated Cl- (ClCa) channels regulates the membrane excitability
and myogenic tone. TMEM16A channels are predominantly form ClCa currents in vascular smooth muscles
including portal vein smooth muscles (PVSMs). Caveola is a cholesterol-rich membrane invaginations andstructurally contributes to effective and efficient signal transduction. Caveolin 1 (Cav1) is accumulated in the
caveolin and plays a key role in forming the functional complex among enzymes, receptors, and ion channels. In
this study, the functional roles of Cav1 on the expression and activity of TMEM16A ClCa channels were
examined in portal vein smooth muscle cells (PVSMCs) from wild-type (WT) and Cav1-knockout (KO) mice.
Spontaneous contractions of PVSMs were recorded using an isotonic transducer. TMEM16A-mediated ClCa
currents were recorded by whole-cell patch-clamp configurations. The expression of TMEM16A channels wasquantitatively analyzed by real-time PCR. The amplitude of spontaneous contractions of PVSMs was larger in
Cav1-KO mice than WT mice. Whole-cell ClCa currents were also larger in Cav1-KO PVSMCs than WTPVSMCs. Importantly, Ani9 (a specific blocker for TMEM16A channels)-sensitive currents were increased in
Cav1-KO PVSMCs compared to WT PVSMCs. The expression of TMEM16A channels was higher in Cav1-KO
PVSMs than WT PVSMs. The present data strongly suggest that the caveola structure formed by Cav1
negatively regulates the expression and activity of TMEM16A-mediated ClCa channels in vascular smooth muscle
cells.

Copyright © The Japanese Pharmacological Society1-B-SS05-6

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The L-DOPA receptor GPR143 in the indirect pathways regulates an anxiety-likebehavior through GPR143-DRD2 coupling〇田近 伶、増川 太輝、内村 放、五嶋 良郎
横浜市立大・院医・薬理We propose that L-DOPA by itself is a neurotransmitter. Recently, a G-protein coupled receptor GPR143, agene product of ocular albinism1, was identified as a receptor for L-DOPA. In this study, to identify thephysiological role of GPR143, we performed phenotypic analysis using Gpr143-gene deficient (GPR143-KO)
mice. To assess anxiety- and exploration-related behaviors, we employed zero-maze test, and found that timespent in open arms was decreased in GPR143-KO mice when compared to wild-type (WT) mice. The timespent in open arms was also decreased in striatal indirect pathway specific GPR143-KO mice. To investigate theinvolvement of endogenous L-DOPA, we examined the effect of alpha-methyl-para-tyrosine, a synthetic
inhibitor of L-DOPA on mouse behavior. We found that administration of α-MPT at the dose of 3mg/kg (i.p.)decreased the release of L-DOPA without affecting that of dopamine from the dorsal striatum. The
administration of α-MPT decreased the time spent in open arms in WT mice, while this effect was notobserved in GPR143-KO mice. Furthermore, intraventricular administration of a synthetic peptide, which
inhibited the interaction between GPR143 and dopamine D2 receptor (DRD2), increased anxiety-like behavior.
These results suggest that L-DOPA regulates anxiety-like behavior through GPR143 and DRD2 coupling in the
striatal indirect pathway.

Copyright © The Japanese Pharmacological Society1-B-SS06-1

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Inhibition of amino acid transporter LAT1 drastically suppresses the transport oflarge neutral amino acids and induces the downregulation of global translationin cancer cells〇西窪 航1、大垣 隆一1,2、岡西 広樹1、奥田 傑3、徐 旻憓1、遠藤 仁4、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研究部門、3東京大・

1

院農学生命科学・食品生物構造、4ジェイファーマ

Nutrient uptake is essential for maintaining the enhanced growth and proliferation of cancer cells. LAT1
(SLC7A5), which preferentially transports large neutral amino acids, is highly expressed in various cancers.
LAT1 inhibitors are preclinically shown to suppress the cancer cell proliferation and tumor growth, and a
representative compound JPH203 is under clinical evaluation. However, detailed pharmacological influence of
LAT1 inhibition on the overall uptake of large neutral amino acids and the protein synthesis in cancer cells thatare thought to be crucial for its anti-cancer effects have not been elucidated yet. Here, we showed that JPH203
dramatically inhibits the uptake of all the large neutral amino acids in multiple pancreatic cancer cell lines. We
also found that JPH203 significantly inhibits the amino acid uptake even in cell culture media containing highconcentrations of various amino acids. Analyses of the protein synthesis activity based on the binding state ofmRNA with ribosomes (Polysome analysis) and the incorporation of puromycin into nascent polypeptides(SUnSET) revealed that JPH203 suppresses global translation. These results advance our understanding ofpharmacological activities underlying the anti-cancer effects of LAT1 inhibitors, further supporting theadequacy of cancer treatments targeting LAT1.

Copyright © The Japanese Pharmacological Society1-B-SS06-2

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Iron tablets delay gastric emptying, which is ameliorated by 5-HT3 receptor
antagonist.

〇山田 舜己1、三原 大輝1、丸山 彩2、水戸部 裕子2、大橋-土井 雅津代2、堀 正敏1、黒澤 珠希1、岸 和寿1、神﨑 萌絵1
1

東京大・農・獣医薬理、2鳥居薬品

Gastrointestinal symptoms, including nausea and vomiting, are common adverse effects of oral iron tablets, but
the mechanism of iron-induced nausea and vomiting is not yet known. Studies have shown that there are close
relationships between gastrointestinal motility and gastrointestinal symptoms such as nausea, vomiting, anddiarrhea, with more than 90% of patients with delayed gastric emptying experiencing nausea and vomiting.
However, the effect of iron on gastrointestinal motility has not yet been investigated. In the present study, we
aimed to elucidate the effects of iron on gastrointestinal motility using sodium ferrous citrate (SFC), the mostcommonly used iron tablets. Gastric emptying in mice was assessed by 13C-octanoic acid breath test to examine
the effect of SFC (3-30 mg Fe/kg, p.o.) with or without the 5-HT3 receptor antagonist, palonosetronhydrochloride (5 mg/kg. s.c.). Colon transit was also measured by the beads method. The results showed thatSFC delayed the gastric emptying, which was ameliorated by administration of palonosetron hydrochloride. It
was also confirmed that ingredients of the tablets had no effect on gastric emptying. SFC also had no effect on
the colon transit. These results lead the possibility that the iron-induced delayed gastric emptying may

be mediated through nausea and vomiting.

Copyright © The Japanese Pharmacological Society1-B-SS06-3

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Regulatory system of cannabinoid type 1 receptors in the basolateral amygdalaon the place preference and anxiolytic-like behaviors〇徳竹 伯洸、浅野 昂、中家 茂俊、宮西 肇、泉尾 直孝、新田 淳美
富山大・大学院医学薬学研究科・薬物治療学研究室

Cannabis is the most widely used addictive drug following alcohol and tobacco. However, the mechanisms
involved in the mental effects and dependence formation are unclear. Δ9-tetrahydrocannabinol (THC), themain active substance in cannabis, binds and affects cannabinoid type 1 receptors (CB1R) in the brain. The
mice wereadministered arachidonylcyclopropylamide (ACPA), a CB1R-selective agonist, and then two

behavioral experiments were performed. Treatments of ACPA induced the anxiolytic-like behavior in theelevated plus maze test. ACPA increased place preference in the conditioned place preference test. The BLA ofmice highly expresses CB1R in the GABAergic interneurons. We aimed to reveal the role of CB1R in BLA for
ACPA-induced behaviors. AM251, a CB1R selective antagonist, was administered intra-BLA before
administration of ACPA. Intra-BLA administration of AM251 inhibited ACPA-induced anxiolytic-like behavior
and place preference. Furthermore, in vivo microdialysis was performed to measure basal GABA levels in the
BLA. Acute administration of ACPA had significantly increased basal GABA levels. Chronic administration of
ACPA didʻt affect basal GABA levels. These results suggest that CB1R in the BLA contributes to behavior
disorders caused by the acute or chronic use of cannabis and these behaviors maight be through a complex
control system involving GABA. This study suggests that CB1R in the BLA may lead to new therapeutic targets
in the treatment of cannabis-induced adverse effects.

Copyright © The Japanese Pharmacological Society1-B-SS06-4

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The involvement of the medial prefrontal cortex in nicotine-induced facilitation of
object recognition memory retrieval〇江崎 博仁1、北中 貴紀2、西谷 直也1,2、出山 諭司1,2、金田 勝幸1,2
1

金沢大・院薬・薬理、2金沢大・薬・薬理We have previously reported that nicotine (Nic) facilitates object recognition memory (ORM) encoding via
activation of excitatory neurons in the medial prefrontal cortex (mPFC) in mice. In this study, we investigatedwhether Nic also facilitates ORM retrieval using the novel object recognition test (NOR). Male C57BL/6J mice
(7 ‒ 12 weeks) received Nic before the test session, which was performed 24 hours after the training session of
the NOR. Systemic administration (0.1 mg/kg; s.c.), but not intra-mPFC infusion (0.3 μg/side), of Nicenhanced ORM, suggesting that Nic facilitates ORM retrieval by acting on brain regions other than the mPFC.
However, suppression of mPFC neuronal activity with inhibitory DREADD hM4Di, which was specifically
expressed in excitatory neurons using an AAV vector, significantly inhibited the systemic Nic-induced
facilitation of ORM retrieval. Moreover, activation of mPFC excitatory neurons with excitatory DREADD
hM3Dq significantly facilitated retrieval of ORM. These data suggest that Nic facilitates ORM retrieval throughthe indirect activation of mPFC excitatory neurons.

Copyright © The Japanese Pharmacological Society1-B-SS06-5

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Involvement of cannabinoid receptor type 2 (CB2) to dendritic cell population on
inflammatory and allergic response〇細木 春花1、野崎 千尋2、古谷 彰子1,3、柴田 重信1
早稲田大・理工・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、3愛国学園短期大学・1

家政科

Cannabinoid receptor type 2 (CB2) is one of the major receptors for cannabis, which is expressed all over the
body especially on the immune-related cells. It is considered that CB2 can work as a regulator in the immune
system, particularly to suppress inflammatory responses in macrophages and microglias when activated.
However no other immune cells have been studied whether and how they get modulated by CB2 activity. The
objective of this experiment is to search for the other immune cell that can be affected by CB2 activity, andreveal the detailed mechanism of CB2-involved immune regulation.
To characterize the CB2-regulated cells under the inflammatory state, we injected LPS (1 mg/kg, i.p.) to CB2
deficient mice (CB2-KO) and wild-type (WT) controls. Spleen has been harvested from these mice 2 hours after
the LPS administration, and immunophenotyping by flow cytometry has been conducted. In this experiment, wefound the particular increase of the type 2 classical dendritic cell (cDC2) population in LPS-treated CB2-KO.
As past studies reported that cDC2 recruits helper T cells when the body is exposed to an allergen, we further
induced allergic rhinitis by OVA sensitization to investigate in-vivo effect of cDC2 increase in CB2-KO and WT
animals. As expected, CB2 deficiency resulted as the significant exacerbation of allergic symptoms compared to
WT mice. These results suggest that CB2 activity may suppress the allergic response by the reduction of cDC2
recruitment. Pharmacological effect of CB2 agonists to these symptoms will be studied as the future experiment.

Copyright © The Japanese Pharmacological Society1-B-SS06-6

学生セッション
2+

A novel T-type Ca channel inhibitor, KTtp38, developed by structuralmodifications of pimozide, a typical antipsychotic agent: Evaluation of thechannel selectivity, electrophysiological characteristics and analgesic activity〇笠波 嘉人1、高島 康宏1、木野 貴博1、石川 千浩2、長南 百香3、豊岡 尚樹2,3、関口 富美子1、坪田 真帆1、川瀬 篤史
4、大久保 つや子5、吉田 繁6、岡田 卓哉2,3、川畑 篤史1
近畿大・薬、2富山大院・理工、3富山大・工・生命工学、4近畿大・薬・生物薬剤、5福岡看護大・基礎・基礎看護、6近畿

1

大・理工・生命科学

In this study, we examined the selectivity, electrophysiological properties and analgesic activity of KTtp38, anovel inhibitor of T-type Ca2+ (Cav3) channels, developed by structural modification of pimozide, a typicalantipsychotic agent. The IC50 value (μM) of KTtp38 was 0.0934 and 1.109 for inhibiting Cav3.2-dependent
currents in response to a test pulse of -20 mV from holding potentials (HPs) of -80 and -110 mV, respectively,
indicating a state dependency. The IC50 of KTtp38 for inhibiting Cav3.1-depedent currents caused by the test
pulse from HP of -80 mV was 0.217 μM. Pimozide, but not KTtp38, at 1 μM completely inhibited the specificbindings of [3H]-spiperone to D2 and D3 receptors in rat striatal membrane fractions. In isolated rat jugular veinrings, the 5-HT2 receptor-mediated contraction was inhibited by pimozide, but not KTtp38, at 10 μM. In mice,i.p. administration of pimozide, but not KTtp38, caused catalepsy. KTtp38 abolished somatic and visceral pain
caused by an H2S donor, known to enhance Cav3.2 activity, in mice. KTtp-38 also reversed oxaliplatin-induced
peripheral neuropathy in wild-type, but not Cav3.2-null, mice. The T1/2 (h) of KTtp38 and pimozide in the blood
was 2.42 and 2.47, respectively. Collectively, KTtp-38 is considered a state-dependent, selective Cav3 inhibitor
and useful as an analgesic.

Copyright © The Japanese Pharmacological Society1-B-SS07-1

学生セッション

Ameliorating effects of corosolic acid in monocrotaline-induced pulmonary
hypertensive rats

〇川出 有希子1、山村 彩2、近藤 るびい1、鈴木 良明1、山村 寿男11

名古屋市大・院薬・細胞分子薬効解析、2愛知医大・医・生理

Pulmonary arterial hypertension (PAH) is a progressive and fatal disease of the cardiovascular system. PAH is
characterized by thickening of the pulmonary artery wall (remodeling) and often causes inflammation aroundthe pulmonary artery. It has been reported that corosolic acid (CRA), is a pentacyclic triterpene acid containedin banaba leaves, has anti-inflammatory, anti-diabetic, and anti-cancer effects. In the present study, the effects
of CRA on the pathogenesis of PAH were examined using monocrotaline (MCT)-induced pulmonary
hypertensive (PH) rats. Male SD rats (4 weeks-old) were injected subcutaneously with vehicle (saline; control)or MCT (60 mg/kg). CRA (1 mg/kg) was administered intraperitoneally daily from 1 week after MCT injection.
At 3 weeks after MCT injection, the effects of CRA on theparameters of PAH pathogenesis wereanalyzed. Hematoxylin and eosin (H&E) staining revealed that CRA clearly improved PAH remodeling in MCTinduced PH rats. The treatment with CRA also reduced the Fulton ratio (an index of right ventricular
hypertrophy) in MCT-induced PH rats. Furthermore, CRA significantly lowered right ventricular systolicpressure (RVSP) in MCT-induced PH rats. In contrast, CRA did not affect these parameters in control rats.
Taken together, CRA may be useful as a novel therapeutic candidate for PAH.

Copyright © The Japanese Pharmacological Society1-B-SS07-2

学生セッション

Honokiol Preserves Mitochondrial Sirtuin 3 And Suppresses Hypoxiareoxygenation Injury in Cultured Myocytes〇谷島 旭、小原 幸、鳥羽 裕恵、中田 徹男
京都薬科大・薬・病態薬科学系臨床薬理学分野[] Honokiol is a small-molecule polyphenol isolated from the genus Magnolia and known as anactivator of sirtuin 3, a mitochondrial deacetylase. We examined whether honokiol attenuates mitochondrial
injury, leading to the attenuation of cell death in hypoxia-reoxygenation (H/R) injury in cardiomyocytes.
[] Neonatal rat cultured cardiomyocytes were subjected to 5 hours of hypoxia followed by

30 minutes of reoxygenation in the presence or absence of honokiol (30 μmol/L). Lethal myocyte injury wasassessed by LDH activity in culture medium and myocyte apoptosis was examined by nuclear staining withDAPI and caspase 3 activity. H/R significantly increased LDH activity and apoptotic myocytes, and treatment
with honokiol significantly attenuated these indices of myocyte death. In mitochondrial apoptotic pathway,
reduction of mitochondrial membrane potential plays critical roles, and ATP is mainly produced by
mitochondria. After H/R mitochondria lost their membrane potential, detected by TMRM fluorescence, leading
to reduction of ATP content in myocytes, and honokiol recovered them. After H/R protein expression of sirtuin
3 was significantly restored by honokiol. Sirtuin 3 is known to deacetylate Mn-SOD. After H/R honokiol
decreased acetylation levels of Mn-SOD and tended to attenuate mitochondrial hydrogen peroxide production.
[] These results indicate that Honokiol protects mitochondria via enhancement of sirtuin 3, leading

to attenuation of H/R-induced myocyte lethal injury.

Copyright © The Japanese Pharmacological Society1-B-SS07-3

学生セッション

Vascular endothelial growth factor inhibitor increases the incidence of aortic
dissection in mice

〇辻中 海斗1,2、石澤 有紀3、大峯 航平1、西 穂香1、吉岡 俊彦1,2、近藤 正輝2、糸数 柊人1、新村 貴博4、相澤 風花2、八木 健太4、合田 光寛1,2、石澤 啓介1,2
1

徳島大・院医薬・臨床薬理学分野、2徳島大学病院・薬剤部、3徳島大・院医薬・薬理学分野、4徳島大学病院・総合臨床研究センター

Aortic dissection is highly lethal, and the risk factors such as hypertension, aging, and atherosclerosis arethought to contribute to its onset. Recently, there has been increasing reports that vascular endothelial growth
factor (VEGF) inhibitors can induce the aortic dissection as an adverse event. However, the association between
VEGF inhibitors and aortic dissection has been unclear. Therefore, we investigated if VEGF inhibitor increases
the onset of aortic dissection using acute aortic dissection model mice (AAD mice).
Sunitinib (100 mg/kg/day) was administered orally for 28days to AAD mice induced by nitric oxide inhibitor,
angiotensin II, and lysyl oxidase inhibitor. Blood pressure was measured every week. After 28days, the incidence
rate of AAD was estimated. For in vitro study, human umbilical vein endothelial cells (HUVEC) were treated bysunitinib for 24 hours. Then, mRNA expressions of intracellular cell adhesion molecule-1 (ICAM-1) andendothelin-1 (ET-1) were measured.
Sunitinib increased systolic blood pressure (182 mmHg vs 288 mmHg with sunitinib ; p＜0.01) and theincidence of AAD (40% vs 59% with sunitinib; p=0.26). Moreover, sunitinib increased mRNA expressions of
ICAM-1 and ET-1 in HUVEC. These results suggested that VEGF inhibitors induced high blood pressure anddeveloped AAD via endothelial damage.

Copyright © The Japanese Pharmacological Society1-B-SS07-4

学生セッション

Eucommia ulmoides oliver leaf extract and geniposidic acid improve hypoxiainduced pulmonary arterial hypertension〇谷 和佳奈1、披田 真里1、松原 匠弥1、田頭 秀章2、喜多 知2、根本 隆行2、平田 哲也3、岩本 隆宏2、喜多 紗斗美1,2
1

徳島文理大・薬・薬理、2福岡大・医・薬理、3小林製薬(株)・中央研究所・研究開発

Pulmonary arterial hypertension (PAH) is a severe and progressive disease that causes right heart failure. The
pathogenesis of PAH is generally characterized by persistent high pulmonary arterial resistance and pulmonary
arterial remodeling. In this study, we investigated the effects of Eucommia ulmoides oliver leaf extract (ELE) on
hypoxia-induced PAH in mice. We observed that Eucommia ulmoides oliver leaf extract (ELE) improve rightventricular systolic pressure (RVSP) and pulmonary vessel muscularization. To identify an active ingredient,geniposidic acid (〜1 mg/kg, 〜5 mg/kg), a major component of ELE, were orally administered to C57BL/6J
mice during exposure to hypoxia for 4 weeks. Geniposidic acid significantly suppressed the elevation of RVSP inhypoxia-induced PAH mice. In addition, hypoxia-induced pulmonary arterial muscularization was slightly
attenuated in geniposidic acid-treated mice. In human pulmonary artery smooth muscle cells (HPASMC),
endothelin-1-induced intracellular Ca2+ elevation was attenuated by geniposidic acid (200 µM). Furthermore,geniposidic acid (50 ‒ 200 µM) increased the maximal respiration in HPASMC. These findings suggest thatgeniposidic acid may be active ingredient of ELE which effectively improve the development of hypoxia-induced
PAH by preventing the vascular remodeling and mitochondrial dysfunction of pulmonary artery.

Copyright © The Japanese Pharmacological Society1-B-SS07-5

学生セッション

Cardiotoxicity assessment of EGFR-TKI using human iPS cell-derived
cardiomyocytes〇柳田 翔太1,2、佐塚 文乃1、林 紗代1、小野 敦2、諫田 泰成1
国立医薬品食品衛研・薬理部、2岡山大・院医歯薬

1【Introduction】Tyrosine kinase inhibitors (TKIs) have improved the survival of patients with various types ofcancer. Growing evidence suggest that cancer therapy-related cardiotoxicity has become important as theserious adverse event associated with many TKIs. Epidermal growth factor receptor-TKI (EGFR-TKI), which
has demonstrated efficacy in patients with non-small-cell lung cancer, has been reported to have a risk of cardiac
dysfunction. However, cardiotoxicity of EGFR-TKIs has not been fully understood. Here we evaluated theeffects of EGFR-TKIs on contractility using human iPS cell-derived cardiomyocytes (hiPSC-CMs).
【Methods】We used iCell cardiomyocyte 2.0 (FCDI). Motion analyses were performed using a cell motion
imaging system (SI8000, Sony). Real-world pharmacovigilance data were analyzed by a reporting odds ratio
from FDA Adverse Event Reporting System (FAERS).
【Results】We found that several EGFR-TKI decreased contraction velocity in a concentration-dependent
manner, while other EGFR-TKIs did not. To confirm thedata, we analyzed the cardiotoxicity risk of

EGFR-TKIs by the real-world pharmacovigilance data from FAERS. EGFR-TKI, which decreased contraction
velocity in hiPSC-CMs, was significantly associated with cardiac failure and decreased ejection fraction.
【Conclusion】Thus, contractile analysis of hiPSC-CMs would be useful to assess TKI-induced cardiac
dysfunction in human. We are planning to evaluate other types of TKIs with hiPSC-CMs.

Copyright © The Japanese Pharmacological Society1-B-SS07-6

学生セッション

Generation of induced pluripotent stem cells derived from a pair of dizygotic
twins.

〇若林 聖士1、長田 千尋1、齋藤 桜子1、坂本 多穗1、山口 賢彦1、砂川 陽一2、諫田 泰成3、森本 達也2、黒川 洵子11

静岡県立大・薬・生体情報分子解析学分野、2静岡県立大・薬・静岡県大薬・分子病態（静岡県立大学 薬学部 分子病

態学分野）、3国衛研・薬理部

In many cardiovascular diseases, differences between women and men have been well described inepidemiology, pathophysiology, clinical manifestations, treatment efficacy, and outcomes, and have attracted
attention as an aspect of personalized medicine. Recently, the use of induced pluripotent stem cells (iPS) cells toreflect individual differences in the drug discovery and the toxicological assay has been attracting attention, but
a screening assay system for human cells to evaluate sex differences has not yet been established. Thus, we here
propose to develop an

system using iPS cell lines derived from a pair of dizygotic twins. We report the

progress of three pairs of clones selected for future sex-differences analysis; no sex differences were observed in
the expression of the three germline differentiation markers. As a next step, we plan to examine sex differencesin gene expression in twin pairs of undifferentiated and differentiated cardiomyocytes to obtain data that will
form the basis for future functional analysis of iPS cell-derived cardiomyocytes, including cardiotoxicity
assessment.

Copyright © The Japanese Pharmacological Society1-B-SS08-1

学生セッション

Evaluation of the efficacy of the mitochondrial mitogen inhibitor Mdivi-1 using
non-alcoholic steatohepatitis (NASH) liver organoids〇望月 まりあ1、田邊 究2、臼井 達哉3、佐々木 一昭41

東京農工大・農・獣医薬理、2東京農工大・農・獣医薬理、3東京農工大・農・獣医薬理、4東京農工大・農・獣医薬理Non-alcoholic steatohepatitis (NASH) is a disease in which fatty liver develops independently of alcohol intake
and progresses to cirrhosis and liver cancer. Currently, no effective treatment for NASH has been found, andnew approaches to elucidate the pathogenesis of the disease are needed. Itʻs also been suggested that NASH isassociated with functional abnormalities in mitochondria, which are involved in lipid metabolism. Ourlaboratory has successfully established liver organoids from NASH model mice that can reproduce the fibrotic
pathology of NASH, and electron microscopic images of NASH liver organoids showed lipid accumulation,
mitochondrial deformation, and aggregation compared to normal liver organoids. Therefore, we analyzed themorphological changes and expression levels of fibrosis-related markers in NASH liver organoids upontreatment with Mdivi-1, a mitochondrial mitogen (DRP1) inhibitor. The results showed that Mdivi-1
suppressed the expression of dendritic morphology in NASH liver organoids and decreased the mRNA
expression levels of Collagen-I and α-SMA. In addition, when Mdivi-1 was administered long-term to mice fed
a NASH-inducing diet, improvement of fatty liver was observed compared to the solvent-fed group. These
results suggest that Mdivi-1 may be useful as a therapeutic agent to improve NASH pathology.

Copyright © The Japanese Pharmacological Society1-B-SS08-2

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Immunoglobulin therapy improves the lysolecithin-induced demyelination ofmouse sciatic nerve via anti-inflammatory macrophage accumulation〇瀬戸口 潔1、林 明子1、河田 紋実1、柳岡 大悟1、笠井 清花1、山口 宜秀1、石橋 智子1、馬場 広子2、大滝 博和1
東京薬科大・薬・機能形態、2新潟医療福祉大・リハビリテーション・作業療法

1

Immunoglobulin (IgG) therapy is a strategy for treatment of autoimmune, immunodeficiency and acuteinfectious diseases. Chronic inflammatory demyelinating polyneuropathy (CIDP) is a rare and refractory
autoimmune disorder of the peripheral nervous system, characterized by symmetric weakness, impaired
sensation and damaged myelin (demyelination). Although intravenous immunoglobulin (IVIg) preparation isused for the therapy, the mechanism of this therapy on demyelination has not been understood. In this study, we
examined the effect of human IgG on the lysolecithin-induced demyelination in the mouse sciatic nerve.
Lysolecithin was injected into sciatic nerves of the ICR mice (day 0) to induce demyelination and 20 mg (i.v.,
day 1) and 10 mg (i.p., day 3) of IVIg preparation or the same volumes of saline for control group wereadministered. Demyelination area and infiltrated macrophages were evaluated with the longitudinal sciatic
nerve sections on the day 7 and 14 by immunostaining. The demyelination areas of the IVIg-treated group weresignificantly less than those of the control group. CD68+ macrophages infiltrated in the lesions and CD68+
CD206+ macrophages were more prominent in IVIg-treated group.
The results suggest that IgG therapy decreased demyelination areas possibly through the accumulation of M2type macrophages.

Copyright © The Japanese Pharmacological Society1-B-SS08-3

学生セッション

Activated protein C suppresses neuropathic pain through activation ofproteinase-activated receptor 1 (PAR1)〇圓尾 賢悟1、池田 裕哉1、坪田 真帆1、王 登莉2、西堀 正洋3、南 達郎4、伊藤 彰敏4、川畑 篤史1
近畿大・院薬・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1

薬研究センター・薬理

The prevention of oxaliplatin-induced peripheral neuropathy (OIPN) by thrombomodulin alfa (TMα) involves
thrombin-dependent activation of protein C (PC) and thrombin-activatable fibrinolysis inhibitor (TAFI), in
addition to inactivation of high mobility group box 1 (HMGB1). We have demonstrated that complement C5a,degradable by activated TAFI (TAFIa), known as carboxypeptidase B (CPB), is involved in OIPN development.
In the present study, we examined the effect of APC on OIPN as well as surgically induced neuropathic pain in
mice, and asked whether proteinase-activated receptor 1 (PAR1) would participate in the effects of APC in
those neuropathic pain models, given that APC is an unbiased or biased agonist of PAR1. The OIPN in mice
was prevented fully by TMα, an anti-HMGB1-neutralizing antibody (HAb) or TAFIa/CPB, and, to a lesser
extent, by APC. Vorapaxar, a PAR1 antagonist, completely and partially canceled the anti-OIPN effects of APC
and TMα, respectively. Interestingly, the neuropathic allodynia caused by partial sciatic nerve ligation was alsoabolished by TMα, and reduced by HAb or APC, and the effect of APC was reversed by vorapaxar. Our datasuggest that PAR1 is involved in the preventive effects of APC and, in part, of TMα on OIPN and surgicallyinduced neuropathic pain.

Copyright © The Japanese Pharmacological Society1-B-SS08-4

学生セッション

Macrophage-Schwann cell communication promotes peripheral nerve
regeneration〇大島 絵莉、林 良憲、篠田 雅路
日本大・歯・生理

Recent studies indicate the importance of the signal relay from macrophages towards Schwann cells in axon
regeneration after peripheral nerve injury. However, the molecular mechanisms underlying axon regeneration
via macrophage-Schwann cell communication remain largely unclear. Here, we explored macrophage-derived
molecules relevant to axon regeneration. After inferior alveolar nerve transection (IANX), the rats showed
hypoesthesia in the lower lip, which was recovered from 10 days after IANX by an intrinsic regeneration
capacity. In contrast, macrophage ablation caused delayed nerve regrowth. Furthermore, c-Jun-positive
Schwann cells, a repair phenotype, disappeared after the removal of macrophages. Cathepsin S (CTSS) frommacrophages promoted recovery from hypoesthesia and cleaved ephrin B2 on fibroblasts. EphB2, a receptor of
ephrin B2, was expressed in Schwann cells. Accelerated recovery from hypoesthesia after IANX following CTSS
treatment was prevented by neutralization of ephrin B2. These results suggest that CTSS from macrophages
liberates ephrin B2 which in turn facilitates axon regeneration in the orofacial regions. Our results lead to thedevelopment of novel therapeutics for hypoesthesia caused by nerve injury targeting CTSS.

Copyright © The Japanese Pharmacological Society1-B-SS08-5

学生セッション

Increased levels of circulating cell-free DNA in COVID-19 patients withrespiratory failure〇若山 勝紀1、中平 毅一1,2、田中 明彦3、福田 陽佑3、趙 晶1、大田 進3、相良 博典3、吉栖 正典1
奈良県立医科大・医・薬理学講座、2Weill Cornell Medicine・Department of Medicine、3昭和大学病院・呼吸器アレル

1

ギー内科

Background: Cell-free DNA (cf-DNA) is known to be released from injured cells and act as a critical activator of
inflammation and the immune system. Patients with COVID-19 could develop respiratory failure and therefore
require oxygen therapy. In this study, we hypothesized that circulating cf-DNA level could reflect the severity of
COVID-19.
Methods: Analyses of cf-DNA levels were performed on serum samples from 95 hospitalized-patients withconfirmed COVID-19 at Showa University Hospital (Tokyo, Japan). Cf-DNA levels were assessed by measuring
the copy number of mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) using quantitative real-time
PCR.
Results: Patients were grouped into Moderate, Severe, and Critical using the severity criteria by the National
Institutes of Health in U.S.A. There was no significant difference on both cf-DNA levels between Moderate andSevere groups, and between Severe and Critical groups. Meanwhile, both of the levels were significantly higher
in Critical group than Moderate group. Patients were also grouped by their respiratory treatment. Both cf-DNAlevels significantly increased in patients with oxygen-supplementation and patients with intubation, comparedto those with no oxygen supplementation and with non-intubation, respectively. There was negative association
between oxygen saturation (SpO2) and cf-nDNA levels, not cf-mtDNA.
Conclusion: These results suggest that serum cf-DNA could serve as a useful biomarker to help determining
therapeutic management for respiratory failure in COVID-19.

Copyright © The Japanese Pharmacological Society1-B-SS08-6

学生セッション

Mild electrical stimulation and heat shock can suppress acute kidney injury (AKI)
to chronic kidney disease (CKD) transition〇津波古 遥希、寺本 啓祐、加世田 将大、Piruzyan Mariam、岩上 長巨、荒川 知南、Suico Marry Ann、首藤 剛、甲斐 広文
熊本大・院医薬・遺伝子機能応用学分野

AKI is considered as a “curable disease”, but recent epidemiological studies and meta-analysis have revealed that
AKI is a risk factor for CKD. Therefore, it is necessary to establish a treatment that can control the AKI to CKDtransition. We have studied mild electrical stimulation (MES) and heat shock (HS; 42℃) that promotes effective
biological responses. Interestingly, in adriamycin (ADR)-induced nephrotic syndrome (NS) mouse model, MES+HS significantly suppressed albuminuria and proteinuria, which are characteristics of NS. We also investigatedthe effects of MES+HS on AKI to CKD transition in a mouse model of bilateral ischemia reperfusion injury (BiIRI). The renal function of Bi-IRI mouse model was rapidly decreased and then recovered over time. However,tubular damage, inflammation and fibrosis were observed even after recovery of renal function. MES+HS
promoted the recovery of renal function in this model. Moreover, MES+HS significantly suppressed tubular
damage, inflammation, and fibrosis, which are indicators of AKI to CKD transition on day 14 after Bi-IRI. It has
been reported that a subpopulation of failed-repair proximal tubular cell (FR-PTC) emerges after AKI and is
involved in the development of chronic disorders. We found that MES+HS reduced the number of Vcam1positive tubular cells, a marker of FR-PTC, suggesting that MES+HS promotes normal tubular repair. Together,
MES+HS can suppress AKI to CKD transition by regulating inflammation, fibrosis and also the emergence of
FR-PTC involved in the chronicity of the renal disorder.

Copyright © The Japanese Pharmacological Society1-B-SS09-1

学生セッション

The involvement of inflammasomes on the learning and memory impairment in amouse model of embolic stroke〇野村 彩衣1、黒川 和宏1、髙橋 浩平1、宮川 和也1、持田（齋藤） 淳美1、武田 弘志2、辻 稔1
1

国際医療福祉大・薬・薬理、2国際医療福祉大・福岡薬・薬理

Post-stroke cognitive impairment (PSCI) is one of the major complications after a stroke and affects quality oflife. Recently, several studies demonstrated that elevated brain proinflammatory cytokines such as TNF-α, IL-6
and IL-1β can induce cognitive impairment. However, the mechanisms underlying PSCI remain unclear. In thepresent study, we investigated whether post-stroke inflammasomes are involved in the development of PSCI
using acetic acid-induced embolic cerebral infarct mice. Long-term learning and memory assessed by thepassive avoidance test was impaired on days 7 and 14 after stroke, whereas short-term learning and memory
assessed by Y-maze test showed no changes. Also, the expression of the phosphorylated AMPA receptor
subunits GluR1 at serine 831 and serine 845 in the dorsal hippocampus were significantly decreased. Under
these conditions, inflammasome-related proteins, including caspase-1, ASC/TMS1, IL-1β, TNF-α and IL-18,
were significantly increased in the dorsal hippocampus 14 days after stroke. The present findings suggest that adecrease in phosphorylated GluR1 at ser831 and ser845 via the inflammasome activation pathway in the dorsal
hippocampus may be involved in the development of learning and memory impairment after embolic stroke.

Copyright © The Japanese Pharmacological Society1-B-SS09-2

学生セッション

Phase-specific synchronization of basolateral amygdalar neurons withneocortical slow oscillations〇佐藤 由宇1、池谷 裕二1,2
1

東京大・院薬・薬品作用、2東京大・Beyond AI研究推進機構

The basolateral amygdala (BLA) shows firing activity synchronized with neocortex slow oscillations, a form of<1-Hz oscillations that occur dominantly during slow-wave sleep or under anesthesia and is believed tocontribute to the formation of emotional memories. Despite its importance for elucidating the neural circuits
involved in the formation of emotional memories, the mechanism of the synchronization has remained unclear,
mainly because it is difficult to record neuronal membrane potentials in deep brain regions, such as the BLA. We
recorded membrane potentials of BLA neurons using a new method that enables whole-cell recording from deepbrain regions in vivo. We found that BLA neurons transiently depolarized at late active phases in the slow
oscillations. To determine the neural source of the depolarization of BLA neuros, we focused on the medial
prefrontal cortex, whose axons projecting to the BLA is known to contribute to the formation of fear memories,
as a candidate region that depolarizes BLA neurons during slow oscillations. BLA-projecting neurons wereretrogradely labeled with channelrhodopsin-2 using retrograde adeno associated virus, and their firing activity
was recorded using an opto-tagging method. These results provide insight into the neural mechanism thatsynchronizes the slow oscillations between the BLA and the neocortex and lead to the elucidation of themechanism underlying the formation of emotional memories.

Copyright © The Japanese Pharmacological Society1-B-SS09-3

学生セッション

Involvement of ceramide kinase and study as a therapeutic target in NiemannPick disease type C〇田内 理紗子、中村 浩之、本田 拓也、内海 直也、橋本 直宏
千葉大・院薬・薬効薬理学(Background and purpose)
Niemann-Pick disease type C (NPC) is a genetic disorder in which patient cells have endosomal/lysosomal
accumulation of cholesterol. No approved drug improves cholesterol accumulation, and the creation of new
therapeutic drugs are desired. Currently, there are many relationships between sphingolipids and NPC havebeen reported. This study investigated the relationship between NPC and ceramide-1-phosphate, produced byphosphorylating ceramide, using NPC1-null mice.
(Results and discussion)
NPC is characterized by clinically affecting the brain and liver; premature death invariably results.
We generated double-knockout (DKO) mice lacking NPC1 and CerK and compared the phenotypes of NPC
mice and DKO mice in these tissues. In the brain, cholesterol accumulation and Purkinje cell survival were
improved in DKO mice compared with those in NPC1-null mice. In the liver, cholesterol accumulation and liver
disorder were improved in DKO mice compared with those in NPC1-null mice. Administration of a CerK
inhibitor to NPC1-null mice delayed the onset of clinical signs and prolonged the lifespan. These results suggestthat CerK may be helpful as a novel therapeutic target for NPC.

Copyright © The Japanese Pharmacological Society1-B-SS09-4

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Hippocampal mitochondrial dysfunction induces anxiodepressive-like behaviors
in mice with neuropathic pain.

〇吉本 夏輝、中島 一恵、中村 庸輝、森岡 徳光
広島大院医系科学・薬効解析

Neuropathic pain (NP) is frequently accompanied by anxiodepressive-like behaviors, yet the mechanisms
remain unclear. Mitochondrial dysfunction induces neuroinflammation and has been implicated in various
neurological diseases, including depression. However, the relationship between mitochondrial dysfunction andanxiodepressive-like behaviors in the NP state is unclear. The current study examined whether mitochondrial
dysfunction is involved in anxiodepressive-like behaviors in mice with NP. NP was induced by partial sciatic
nerve ligation (PSNL) of male ddY mice. Anxiodepressive-like behaviors were evaluated by forced swim test,social interaction test, and novelty suppressed feeding test. Mitochondrial dysfunction was assessed byquantifying mitochondrial DNA in the cytoplasmic fraction. The expression of type Ⅰ interferon mRNA was
analyzed by real-time PCR. Curcumin was orally administered to inhibit mitochondrial dysfunction. PSNL
induced anxiodepressive-like behaviors with accompanying mitochondrial dysfunction and increase of type I
interferon mRNA in the hippocampus at 8 weeks post-injury. Curcumin suppressed mitochondrial dysfunction
and improved anxiodepressive-like behaviors. The current study suggests that mitochondrial dysfunction in the
hippocampus could be involved in anxiodepressive-like behaviors under NP state.

Copyright © The Japanese Pharmacological Society1-B-SS09-5

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Ceramide kinase, a lipid-metabolizing enzyme, is involved in thepathophysiology of schizophrenia and Parkinson's disease by regulating thelevels of extracellular dopamine.

〇金原 拓哉、安江 匡隆、本田 拓也、中村 浩之
千葉大・院薬・薬効薬理学

Dopamine (DA) is a critical neurotransmitter which modulates motor functions, learning and motivation.
Abnormal DA signaling is related to neuronal diseases such as schizophrenia (SZ) and Parkinsonʻs disease(PD). The symptoms of SZ are classified as hyperDAergic positive symptoms and hypoDAergic negative
symptoms. PD is a common progressive neurodegenerative disorder characterized by the loss and degradationof DAergic neurons. Current drugs poorly treat both diseases.
Ceramide kinase (CerK) is an enzyme which phosphorylates ceramide, a central metabolite of sphingolipids to
produce ceramide-1-phosphate (C1P). CerK/C1P pathway is reported to be involved in extracellular
homeostasis of some neurotransmitters, however, the involvement with SZ or PD remains unclear.

In this study, we created SZ and PD model in mice genetically deletedwith wild-type mice. We found thatand compared their phenotypes

deficiency exacerbated the positive symptoms of SZ partly due to anincrease in the extracellular DA levels. In contrast, negative symptoms of SZ and motor dysfunction of PD werepartly improved by

deficiency.

These results suggest that CerK may be a new therapeutic target for hypoDAergic diseases such as negative
symptoms of SZ and PD by regulating extracellular DA levels.

Copyright © The Japanese Pharmacological Society1-B-SS09-6

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Identification of two distinct neuronal subpopulations encoding parenting andaggressive behaviors toward pups in the population of amygdalohippocampal
area neurons projecting to the medial preoptic area using projection-specific andactivity-dependent labelling〇佐藤 圭一郎、南 雅文、天野 大樹
北海道大・院薬・薬理

Although parenting behavior is necessary for development of infants, underlying neural mechanisms remain
unclear. The amygdalohippocampal area (AHi) neurons projecting to the medial preoptic area (MPOA), a key
region for parenting, were shown to be activated by both parenting and aggression toward pups using retrograde
tracer and c-Fos immunostaining. We labeled MPOA-projecting AHi neurons in a projection-specific andactivity-dependent manner using a retrograde adeno-associated virus vector expressing Cre recombinase
activity-dependently and mice expressing tdTomato Cre-dependently. As a result, we observed the presence of
two functionally distinct subpopulations of parenting and aggressive response neurons.
We next performed scRNA-seq to determine whether the two populations were molecularly distinct and found
395 genes exhibiting higher expression levels in the parenting subpopulation than in the aggression
subpopulation, and 755 genes showing the opposite pattern. Among them, we focused on 5-HT7 receptor
because of high expression ofin the parenting subpopulation. Intraperitoneal administration of LP44, 5-

HT7 receptor agonist, resulted in activation of parenting-labeled, but not aggression-labeled, neurons. Finally,
microinjection of LP44 into the AHi 15 min before behavioral test promoted parenting behavior.
In conclusion, we identified distinct subpopulations of MPOA-projecting AHi neurons encoding parenting andaggressive behaviors toward pups. The results demonstrated that parenting AHi neurons expressed 5-HT7
receptors and were activated by administration of 5-HT7 receptor agonist, which suppressed aggressive
behavior toward pups.

Copyright © The Japanese Pharmacological Society1-B-SS10-1

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Arcadlin induction affects dendritic spine density in the hippocampal dentate
gyrus after cerebral ischemia〇中澤 秀真、井上 耀介、井上 翔太、山口 菜摘、中谷 仁、澤野 俊憲、田中 秀和
立命館大・院生命科学・薬理学研究室

Dendritic spine morphological changes occur in the brain after cerebral ischemia. Arcadlin, a non-clusteredprotocadherin δ2, is induced in a neuronal activity and reduces the dendritic spine density via endocytosis of
N-cadherin. Cerebral ischemia induces neuronal activity but the expression of Arcadlin and its role in theischemic brain are not clear. In this study, we analyzed Arcadlin expression and dendritic spine changes after
cerebral ischemia using a highly reproducible mouse model of middle cerebral artery occlusion (MCAO). We
found thatmRNA was significantly upregulated in the hippocampal dentate gyrus (DG) at 4 hours

after MCAO. Dendritic spine density in the ipsilateral DG was lower than in sham mice. These results suggestthat Arcadlin may be involved in the reduction in the dendritic spine density. We are performing the comparisonof dendritic spine density between MCAO and Sham using

KO mice.

Copyright © The Japanese Pharmacological Society1-B-SS10-2

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–/–Dendritic morphology of the pyramidal cells in the piriform cortex of Arcadlinmice〇北川 貴士1、河前 なつみ1、重松 成秋1、高山 晃行1、上村 健士郎1、山形 要人1、杉浦 弘子2、澤野 俊憲1、中谷 仁1、
田中 秀和1
1

立命館大・院生命科学・薬理学研究室、2東京都医学総合研・脳・神経科学研究分野

Arcadlin (Acad/Protocadherin-8) is a non-clustered d2-protocadherin of the cadherin superfamily. Acad is
induced quickly by neural activity and is known as a molecule that reduces spine density in the hippocampal
neurons in vivo and in vitro. Pyramidal neurons in the layer II of the piriform cortex strongly expresses
mRNA. The piriform cortex is often kindled electrically for seizure generations. We hypothesize that the neuralactivity of the piriform neurons induces Arcadlin, which in turn modulates their spine morphology. In order toinvestigate this possibility, we examined the dendritic morphology of the pyramidal neurons in the piriform
cortex in

mice. Contrary to our expectation,

mice showed a lower spine density than WT micein the pyramidal cells of piriform cortex. The change in spine density was most obviously observed in thinspines, and in the dendritic zone distal to the cell body. The data suggest that Acad does not suppress thepiriform pyramidal cells at least in unkindled status. We will further examine whether the induced Acad
suppresses the piriform pyramidal cells under kindled condition.

Copyright © The Japanese Pharmacological Society1-B-SS10-3

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Elucidation of the pathogenesis of short-term memory impairment in a mousemodel of lipopolysaccharide-induced inflammation〇森本 康平、江口 遼太、乙黒 兼一
北海道大・院獣医

Diseases with peripheral inflammation, such as sepsis and peritonitis, are associated with an increased risk ofcentral nervous system diseases. Injection of lipopolysaccharide (LPS) into mice has been widely used as adisease model for peripheral inflammation and exhibit cognitive dysfunction. Although glial cells have been
implicated in the pathogenesis of cognitive dysfunction, the detailed mechanisms remain unclear, and moststudies have been conducted in young or old mice. In this study, we examined the effects of LPS on short-term
memory in adult mice to elucidate the mechanisms of pathogenesis.
C57BL/6N mice (male, 11-13 weeks old) were injected with LPS (3 mg/kg, i.p.). At 7 days after injection, thenovel object recognition test was conducted, and LPS decreased the discrimination index. At 7 days after
injection, the number of c-Fos+ cells, a marker of neuronal activation, decreased in the hippocampal CA1
region, and the percentage of immature spines detected by Golgi-Cox staining was increased. The gene
expression of several inflammatory factors peaked at 1-3 days after LPS injection and recovered to pre-injectionlevels at 7 days after injection.
These results indicate that the short-term memory impairment was observed at 7 days after LPS injection.
Inhibition of hippocampal CA1 neuron activation and the decrease in synaptic strength due to immature spines
may be involved in the short-term memory impairment.

Copyright © The Japanese Pharmacological Society1-B-SS10-4

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Copyright © The Japanese Pharmacological Society1-B-SS10-5

学生セッション

What factors contribute to diosgenin-induced memory recovery?

〇永田 朋也、楊 熙蒙、東田 千尋
富山大・薬・和漢医薬学総合研究所 神経機能学領域

Alzheimerʻs disease (AD) is a neurodegenerative disorder characterised by deposition of Aβ andhyperphosphorylated tau in the brain. In addition, hyperactivation of glial cells, cerebrovascular damage, andstructure changes of white matter are also observed in AD brain. We previously found that diosgenin
regenerated axons in the brain and improved memory impairment in AD model mouse (5XFAD). However,other beneficial effects of diosgenin leading to memory recovery remain unknown. This study aimed toinvestigate the effect of diosgenin on morphology of glial cells, blood vessels, and myelin in 5XFAD mousebrains.
Vehicle solution or 0.1μmol/kg diosgenin were orally administered to wild-type and 5XFAD mice for 14 days.
Diosgenin administration significantly improved object recognition memory in 5XFAD mice. The brain slices
were served for immunohistochemistry. Diosgenin administration did not remarkably influence the areas of
GFAP+ astrocytes, Iba1+ microglia and CD31+ blood vessels at least in the prefrontal cortex, hippocampus, andperirhinal cortex in 5XFAD mice. We are currently evaluating myelin formation and neurogenesis.
This study indicated that diosgenin administration didnʻt provide no numerical changes of astrocytes, microglia,
and blood vessels in the 5XFAD mouse brains. Therefore, we are now narrowing down the most contributing
factor to memory improvement by diosgenin.

Copyright © The Japanese Pharmacological Society1-B-SS10-6

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Food-derived amino acid ergothioneine inhibits histamine metabolizing enzyme
and promotes anti-inflammatory M2 microglia polarization〇上田 優花、石本 尚大、勝部 諒、増尾 友佑、加藤 将夫
金沢大・薬学系

Oral administration of a food-derived amino acid ergothioneine (ERGO) enhances cognition in mice althoughmolecular mechanisms remain unclear. Our comprehensive molecular targeting assays showed inhibition by
ERGO of histamine N-methyltransferase (HNMT), a main histamine-metabolizing enzyme in the brain (JFunct Foods 95, 105165, 2022). We here attempted to characterize the inhibitory effect of ERGO on HNMTinduced methyl transfer from a methyl donor S-adenosyl-methionine (SAM) to histamine. A radioenzymatic
assay using a [3H]SAM showed that mouse brain homogenate increased radioactivity of the [3H]Nmethylhistamine, while incubation with ERGO or an HNMT inhibitor metoprine significantly suppressed theradioactivity, suggesting that ERGO inhibits murine brain HNMT. Lineweaver-Burk analysis using humanrecombinant HNMT showed that the inhibition by ERGO was competitive with histamine. Quantification of Nmethylhistamine by LC-MS/MS showed that human recombinant HNMT produced N-methylhistamine in atime-dependent manner, whereas ERGO and metoprine significantly suppressed the production. We thenexamined the activation of microglia, immune cells in the brain, because histamine is an important molecule in
the immune system. Immunochemical analysis showed that HNMT was expressed in primary cultured microglia(PCM). Exposure of PCM to ERGO or metoprine significantly increased mRNA expression of CD206, a marker
for anti-inflammatory M2 microglia. Thus, the HNMT inhibition by ERGO may be associated with microglial
polarization in the brain.

Copyright © The Japanese Pharmacological Society2-B-SS01-1

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Inhibitory effect of mirogabalin for various pruritogens-induced acute itch

〇堀 圭汰1、澤幡 雅仁2、久米 利明2、歌 大介2
富山大・院医薬・応用薬理、2富山大・院医薬・応用薬理

1Itch is defined as an irritating sensation that triggers a desire to scratch. The signaling pathways of itch fall into
two main categories, histaminergic and non-histaminergic pathway. Mainly, C fibers transmit itch informationin both pathways. Since it is known that α2δ subunits composing voltage-gated Ca2+ channels are expressed in
C fibers, mirogabalin, a novel gabapentinoid that targets these subunits, may be applicable as an antipruritic
drug. In this study, using male ICR mice, we showed effect of mirogabalin on scratching behavior induced byseveral pruritogens (histamine, chloroquine and compound 48/80). Scratching bouts increased by thesepruritogens are decreased by oral administration of mirogabalin (10 mg/kg). The oral administration ofmirogabalin (10 mg/kg) exhibited no sedation. In addition, the scratch behavior was inhibited by intracisternal
injection of mirogabalin, but not local intradermal injection. These results suggest that mirogabalin is effective
against itch transmitted through both histaminergic and non-histaminergic pathway and also that central nerve
system, especially spinal cord, are involved in the antipruritic effect of mirogabalin.

Copyright © The Japanese Pharmacological Society2-B-SS11-1

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Lysophosphatidic acid (LPA) improves glomerular histology in a murine model ofSLE〇永田 亘、小泉 明穂、中川 慶一、髙橋 さやか、石塚 俊晶
防衛医科大・院医・薬理学講座

Objective: Lupus nephritis is a typical clinical manifestation of SLE. We previously reported that LPA treatment
improves depressive-like behavior and microglial activation in MRL/lpr mice (an animal model of SLE). Thus,
we examined the eﬀects of LPA on renal function and glomerulonephritis in MRL/lpr mice. Method：18-weekold MRL+/+ mice (n=12) were treated with vehicle and 18-week-old MRL/lpr mice (n=24) were treated withvehicle or LPA (1 mg/kg) for 2 weeks. After the treatment, urine and blood samples were collected, andhistological examinations were performed. Results and Discussion: The significant increases in daily urinary
albumin levels in MRL/lpr mice were lost by LPA treatment. Creatinine in plasma was not significantly different
between the three groups. The significant increases in plasma dsDNA antibody levels in MRL/lpr mice were lostby LPA treatment. The increases of CD68-positive cells in the glomerulus were found in MRL/lpr mice and theincreases were suppressed by LPA treatment. The PAS-positive rates in MRL/lpr mice were significantly
increased compared to MRL+/+ mice and the increases were significantly suppressed by LPA treatment. These
results suggest that LPA treatment improves glomerulonephritis and proteinuria in MRL/lpr mice.

Copyright © The Japanese Pharmacological Society2-B-SS11-2

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The efficiency of leucine administration on the maintenance of systemic immune
function in a sarcopenia model〇望月 万莉井、細田 佑樹、稲葉 奏介、劉 爽、茂木 正樹
愛媛大・院医・薬理学

It is well known that chronic inflammation causes the sarcopenia. Whether the efforts to prevent skeletal muscle
loss benefit the immune homeostasis remains unclear. In the present study, the efficacy of leucine
supplementation on the maintenance of systemic immune function was evaluated. A sciatic nerve denervationinduced sarcopenic model was established and the volume and thickness of skeletal muscle on the hindlimb were
evaluated by magnetic resonance imaging. Oral administration of leucine was carried on for 56 weeks. The
skeletal muscle mass and the subsets or function of immune cells were analyzed.
In leucine-treated sarcopenic mice, skeletal muscle mass on the hindlimb was significantly increased comparedto that in non-treated sarcopenic mice. Leucine treatment repaired the mitochondria dysfunction in splenocytes
from sarcopenic mice bothand. In sarcopenic mice, an increase of the PD-1 expression wasobserved in CD4+ and CD8+ T cells. However, oral leucine administration restored the expression of PD-1 inthe lymphocytes to the level of non-sarcopenic mice.
In conclusion, the administration of leucine exhibits beneficial effects on sarcopenia and may influence the anticancer immune responses via adaptive immune resistance mechanism.

Copyright © The Japanese Pharmacological Society2-B-SS11-3

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Roles of CCR5 in development of fibrosis in severe asthma〇霜良 勇人、長谷 雪乃、羽口 天太、髙森 伊富、松田 将也、北谷 和之、奈邉 健
摂南大・薬・薬効薬理学研究室

Mechanisms underlying development of lung fibrosis in severe asthma have been unclear. In our murine modelsof mild and severe asthma, lung fibrosis was significantly developed only in the severe asthma model. RNA-seq
analyses revealed that not only genes of collagen type 1 but also genes of matrix metalloproteinases (MMPs) andactivins, TGF-b family cytokines were found to be highly expressed in the lung of the severe asthma modelcompared with that of the mild model. More interestingly, pathway analyses showed that pathway of CCR5 andthe ligands were up-regulated in the severe model. In this study, roles of CCR5 in the development of lungfibrosis in the severe model were analyzed. Treatment with a CCR5 antagonist, maraviroc but not
dexamethasone exerted significant inhibition of the development of lung fibrosis. Interstitial macrophages (IMs)that expressed CCR5 were markedly increased in the lung, and the degree was significantly higher in the lung ofsevere asthma model than that of the mild asthma model. Real-time RT-PCR analyses revealed that IMs derived
from the lung of severe model expressed higher mRNA levels of MMPs and activins than those of mild asthma
model. From these results, it was strongly suggested that CCR5+ IMs were possibly involved in the developmentof the lung fibrosis in severe asthma.

Copyright © The Japanese Pharmacological Society2-B-SS11-4

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Inhibitory effect of selective serotonin reuptake inhibitors (SSRIs) on Toll likereceptor-dependent and -independent production of IL-6

〇竹中 洋平1、田中 隆2、北畠 和己1、倉持 幸司2、青木 伸1、月本 光俊1
1

東京理科大・院薬、2東京理科大・院理工

Inflammatory diseases lead to excessive proinflammatory cytokine production (cytokine storms). The cytokine
storms are highly lethal, so establishing an effective treatment is desirable. Recently, some of selective serotonin
reuptake inhibitors (SSRIs), which are used to treat depression, have been reported to be effective in treating
various inflammatory diseases, such as COVID-19. In this study, in order to elucidate which SSRI would be themost suitable as an anti-inflammatory drug, we investigated that the effect of 5 SSRIs on the production of
inflammatory cytokine (Interleukin-6; IL-6) induced by macrophage activation induced in a Toll-like receptor(TLR)-dependent manner, and by T cell activation induced in a TLR-independent manner. In J774.1 murine
macrophage cells, pretreatment with the SSRIs significantly suppressed IL-6 release induced by TLR3, TLR4,
and TLR9 agonist. On the other hand, these SSRIs are also significantly suppressed IL-6 release induced by T
cell activator in murine splenic lymphocytes. Our results show that fluoxetine has potent inhibitory effect on IL-6 production induced by various stimuli and low cytotoxicity among the 5 SSRIs. An examination of thestructural requirements indicated that the-methyl group of fluoxetine has a critical role in the inhibition of IL-6 production. Overall, our findings suggest that fluoxetine might be one of the preferred SSRI for further
evaluation as an anti-inflammatory drug to treat cytokine storms.

Copyright © The Japanese Pharmacological Society2-B-SS11-5

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Activation of regulatory T cell through prostaglandin E2-EP4 signaling〇松浦 竜真、Thumkeo Dean、成宮 周
京都大・院医・創薬医学講座

Prostaglandin (PG) E2, a bioactive lipid biosynthesized from arachidonic acid, exerts its functions through 4
cognate receptors EP1-4. Our previous studies suggested that PGE2 attenuates antitumor immunity in thetumor microenvironment by the recruitment and activation of regulatory T cells (Tregs), which are a subset of T
cells specialized in immunosuppression. However, whether such actions of PGE2 on Treg is a direct or indirect
action remains largely unknown. To address this question, we generated induced Tregs (iTregs) from purified
splenic CD4 + T cells by CD3/28 stimuli in the presence of TGF-β

and examined the direct effect of

PGE2. Using flow cytometry analysis, we found that the expression level of Foxp3, a master transcription factorof Treg, and 4-1bb, a coactivator of Treg, were both significantly enhanced upon PGE2 treatment. Furthermore,these effects were notably suppressed in the presence of EP4 antagonist, suggesting that PGE2 contributes toTreg activity through the EP4 receptor. In addition, RNA-seq revealed that not only
coactivator genes such as

,

, and

, but other Treg

were greatly increased in the PGE2 treatment group. Given that

previous studies reported that high expression level of these genes in intratumoral Tregs is positively correlated
to Treg immune suppression activity, we speculated that PGE2 produced in the tumor microenvironment may
directly induce the activated phenotype of intratumoral Treg through EP4 receptor.

Copyright © The Japanese Pharmacological Society2-B-SS11-6

学生セッション

The effects of dasatinib on muscle regeneration〇石田 七海1、黒澤 珠希2,3,4、後藤 もも2、堀 正敏2
東京大・農・獣医薬理学教室、2東京大・院農学生命科学・獣医薬理、3徳島大・院医歯薬・生体栄養学分野、4文部科学

1

省・日本学術振興会・特別研究員PDAs there are no effective treatments for muscular dystrophy (MD), identifying systemically acting smallmolecule therapeutics is desirable. Tyrosine phosphorylation of β-dystroglycan, which occurs via tyrosine
kinase in dystrophin-deficient muscles, has been reported to induce muscle damage, and several tyrosine kinase
inhibitors have been researched as potential therapeutic agents for MD. Nilotinib, a second-generation tyrosine
kinase inhibitor, was potentially effective in MD by reducing muscle fibrosis. However, there was a problem thatits direct effect on satellite cells inhibited muscle differentiation. Dasatinib, a third-generation tyrosine kinase
inhibitor, is also expected to be effective in MD, but its effect on muscle regeneration is unknown.
Here, we investigated the effects of dasatinib on muscle, with a focus on muscle regeneration. We administereddasatinib to mice whose tibialis anterior muscle was damaged by cardiotoxin. In the dasatinib-treated mice,abnormal myofibers were observed, and muscle regeneration may be impaired. The effect on muscle
differentiation was examined using the myoblast cell line C2C12. There was abnormal cell fusion, and thisabnormality differed from that previously described for nilotinib. Further research is currently underway intomechanisms causing the abnormal muscle regeneration.

Copyright © The Japanese Pharmacological Society2-B-SS12-1

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Mechanisms for anti-apoptotic effects of nobiletin in pancreatic β-cells〇梶 萌、金子 雪子、石川 智久
静岡県立大・院薬・薬理

The chronic hyperglycemia in type 2 diabetes causes deterioration of pancreatic β-cell dysfunction
due to decreases in insulin secretory response and β-cell mass. Nobiletin, a citrus flavonoid, has beenreported to improve hyperglycemia and insulin resistance in type 2 diabetic model mice. We
previously showed that nobiletin, a citrus flavonoid, exerts insulinotropic and anti-apoptotic effects
in pancreatic β-cells through the elevation of cAMP levels. In the present study, we investigatedmechanisms for anti-apoptotic effects of nobiletin in INS-1 cells, a rat β-cell line. Endoplasmic
reticulum stress was induced by thapsigargin, tunicamycin, or chronic high glucose exposure. The
expression of apoptosis-related proteins in INS-1 cells was analyzed by Western blotting. Nobiletinsignificantly suppressed the elevation of cleaved caspase-3 expression induced by these proapoptotic
stimulations. The expression of thioredoxin interacting protein (TXNIP), a regulator of cellularoxidative stress, was also suppressed by nobiletin treatment. Nobiletin slightly restored the decreasein phosphorylated Akt levels induced by thapsigargin or tunicamycin treatment. These results
suggest that nobiletin suppresses apoptosis induced by endoplasmic reticulum stress via thedegradation of TXNIP, which might be mediated by Akt.

Copyright © The Japanese Pharmacological Society2-B-SS12-2

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Efficacy of an alkali extract of Sasa sp. in a mouse model of acute kidney injury〇小関 優太朗1,2、柴田 佳太1,2、藤澤 知弘3、野部 浩司1,2
1

昭和大・院薬・薬理、2昭和大・薬理科学研究セ、3大和生物研

Acute kidney injury (AKI) is a catastrophic disease with high morbidity and mortality in hospitalized patientsand contributes to the pathogenesis of chronic kidney disease. However, there are no approved effective
treatment for AKI. There are many causes of AKI, including ischemia, hypoxia, and nephrotoxicity. The primarycause is ischemia reperfusion injury, due to trauma, shock, sepsis, and renal transplantation. Previous studies
have shown radical scavenging activity and anti-inflammatory activity of an alkali extract ofstudy aimed to evaluate the efficacy of an alkali extract of

sp. The present

sp. in AKI model mice. AKI was induced bytemporary vascular clamping of the left kidney for 45 min followed by reperfusion, two weeks after removal ofright kidney. We have measured the renal function using urine and serum, and morphological assessment. The
levels of inflammation markers in the kidney have also been measured by ELISA. An alkali extract ofsp.

improved the renal function, the damage of renal tubules and exhibited anti-inflammatory in the kidney. Thesefindings indicate that an alkali extract ofsp. protects kidney in AKI model mice.

Copyright © The Japanese Pharmacological Society2-B-SS12-3

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Effects of Cistanche tubulosa extract on cervical spondylotic myelopathy〇須山 真聡、東田 千尋
富山大・院医薬・薬科学専攻

In cervical spondylotic myelopathy (CSM), degenerative changes affecting the vertebrae, intervertebral disks
and ligaments compress the spinal cord. Subsequent neuronal damage leads to motor and sensory dysfunctions.
Although standard treatment of CSM is surgical decompression, neurological impairment sometimes remains orrecurs. Therefore, fundamental therapy that recovers neuronal damage is required.
(CT) extract improved motor dysfunction in spinal cord

Our previous study clarified thatinjury. Since we supposed that the extract might be effective also for CSM, this study aimed to investigate
therapeutic effects of CT extract on CSM.
30% ethanol CT extract was orally administered to CSM model mice. Motor and sensory functional changes
were observed. Recovery of spinal axons were also evaluated by visualization using neuronal tracers. To clarify
functional mechanism of CT extract to axons, we focused on acteoside and echinacoside that are main
components in CT extract. After oral administration of CT extract, echinacoside transferred to the spinal cordand brain. Acteoside also transferred to the spinal cord. Therefore, we investigated effects of these components
on primary cultured cortical and spinal neurons. Acteoside increased axonal and dendrite densities.
This study showed that acteoside transferred to the spinal cord after oral administration of CT extract, andextended axons. Experiments of CT extract on functional recovery and axonal repairing in CSM model

mice are now under investigating.

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Metabolic changes in hypothalamic glial cells under pathological conditions ofcancer cachexia〇渡邊 諒1、須田 雪明1,2、成田 道子2,3、松山 富貴子1,4、濱田 祐輔1,2、葛巻 直子1,2、成田 年1,21

星薬科大・薬・薬理、2国立がん研セ・研・がん患者病態生理、3東京医科大・医学総合研・分子細胞治療、4日本小動物

医療セCancer cachexia affects many patients with terminal cancer. It significantly reduces their quality of life and also
affects their survival rate. Biological homeostasis helps the body adapt to stimuli such as stress andenvironmental changes, and a breakdown of this control mechanism is thought to lead to the development ofvarious diseases. To better understand the fluctuations of biological homeostasis in these disease pathologies, it
is important to understand cellular metabolomics. In this study, we focused on metabolic changes inhypothalamic glial cells under conditions of cancer cachexia. As a result, we found that the expression of several
mRNAs involved in glycolysis and the TCA cycle were markedly altered in glial cells isolated from the
hypothalamic region of cancer cachexia model mice. Based on these results, we speculate that metabolic
abnormalities may be induced in hypothalamic glial cells under the pathological condition of cancer cachexia.
We are currently investigating the details by a comprehensive metabolomic analysis.

Copyright © The Japanese Pharmacological Society2-B-SS12-5

学生セッション

Elucidation of interaction factors between COPD and lung cancer〇林 恵1、中嶋 竜之介1、髙橋 宜暉1,2、岸本 朋樹1、福山 絢美1、小笠原 長耀1、スイコ メリーアン1、甲斐 広文1、首藤 剛1
1

熊本大・薬・遺伝子機能応用学分野、2熊本大・院医薬・博士課程リーディングプログラム「HIGO」

Chronic obstructive pulmonary disease (COPD) is the third leading cause of death in the world and ischaracterized by inflammation, emphysema and respiratory dysfunction. The development of radical treatment
and the achievement of long-term disease management are urgent issues for COPD. Because COPD is highly
complicated with lung cancer and this complication leads to poor prognosis, it is very important to elucidate its
pathological mechanism and develop a novel therapeutic strategy. Previous research showed the relationshipbetween lung cancer and COPD-derived inflammation, but not emphysema. Therefore, I selected elastase
model and induced lung cancer using tobacco-specific carcinogenesis (NNK) for lung cancer initiation stage
model (COPD-NNK) and lewis lung carcinoma (LLC) for lung cancer exacerbation stage (COPD-LLC). In
COPD-NNK model, incidence of lung tumors was increased but COPD phenotypes were not exacerbated.
Moreover, I found that α7nAChR-p-Akt pathway was activated in these tumors. Additionally, the survival rate
was decreased and intratumor T cells and immune checkpoint protein PD-L1-positive macrophages wereincreased in COPD-LLC model. In contrast, the survival rate was increased in anti-PD-L1 antibody-treated
COPD-LLC model. These results suggested that PD-L1-positive macrophages inhibited T cell activity anddecreased the survival rate. In this study, I revealed changes in tumor microenvironment and exacerbation
mechanism in COPD-lung cancer complication.

Copyright © The Japanese Pharmacological Society3-B-SS13-1

学生セッション

Establishing a mouse model of cervical spondylotic myelopathy and effect ofacteoside on neurological disabilities in the model〇羽柴 圭悟、東田 千尋
富山大・和漢研

Cervical spondylotic myelopathy (CSM) is the most common cause of neurological disability in worldwide,
which is caused by chronic compression of the spinal cord. Mainstay of treatment is surgical decompression,conservative treatment, or symptomatic analgesics. However, many patients end up with substantial residual
disability. Disabilities of motor and sensory function are caused by neuronal damage and axonal loss. Therefore,
we supposed that accomplishment of axonal growth is the most critical therapeutic strategy for CSM. Since ourprevious study suggested acteoside as a facilitator of axonal growth in spinal cord injury, this study aimed toinvestigate effects of acteoside on functional disability in CSM model mice.
The animal models of CSM reported so far have experimental disadvantages, such as difficulties of controllingcompressive intensity, and unstable and too long time-course for appearance of motor dysfunction. First, weestablished a new mouse model of CSM to solve the problems. After laminectomy at C3-C5, the cervical cord
was compressed by a mini screw. Motor functions of forelimbs and hind limbs decreased depth-dependently of
the compression. The dysfunction was obvious at least 9 days after the compression. At the compressed center,
GFAP-positive astrocytes increased, and NF-H positive axonal density decreased compared to sham mice. Since
we succeeded establishing CSM model, acteoside was administered orally from 7 days after compression. Motorand sensory functions and histological evaluation are now under investigation.

Copyright © The Japanese Pharmacological Society3-B-SS13-2

学生セッション

Effective drug for optic nerve growth in a mouse model of normal tension
glaucoma and it‘s mechanism〇渋江 省吾、東田 千尋
富山大・和漢医薬学総合研究所・神経機能学領域

Glaucoma is a major cause of irreversible blindness worldwide, and is evoked by degenerationand loss of retinal ganglion cells (RGC). Current drug treatments are focused on lowering intraocular pressure,which donʻt accomplish vision recovery. Our laboratory previously found axonal regeneration activity of Drug A(name is closed due to the patent). Therefore, this study aimed to investigate effects of Drug A on optic nerve
growth in an optic nerve crush model.
Based on our previous data showing that intravitreal injection of Drug A increased optic nerve density in the
optic nerve crush model, this study aims to investigate oral treatability and potency of Drug A.
At first, we evaluated brain penetration of Drug A after p.o. administration. Drug A was detected in the retina,
the optic nerve and whole brain at least 6h after p.o. administration.
Immediately after optic nerve crushing, Drug A or vehicle solution was administered orally for 3 weeks.
Intraocular pressure measurements and a behavioral vision test were conducted on the day of sacrificing. After
that, the retina, optic nerves and brain were dissected and served for histochemical evaluation. The number ofretinal ganglion cells and optic nerve termination to the lateral geniculate nucleus were increased by Drug A.
Drug A treatment to primary cultured RGC for 4 days significantly enhanced axonal length. The mechanism ofDrug A for axonal growth of RGC is under investigation.

Copyright © The Japanese Pharmacological Society3-B-SS13-3

学生セッション

Sleep homeostasis of a monophasic sleep in reptiles〇羽鳥 聖七、山口 翔、松井 双葉、周 至文、乘本 裕明
北海道大・医・細胞薬理学教室

Sleep is essential for human beings to live a healthy life and to build a healthy society. However, there are few
efficient drugs or treatments for sleep disorders since the detailed neuronal mechanisms associated with sleep
are unknown. In particular, monophasic sleep at night as human is less understood due to a lack of modelorganisms. For example, most rodents used for life science research are nocturnal and take a polyphasic sleep.
Recently, we introduced a reptile, Australian bearded dragon
sleep research.

as a new model organism foris diurnal and takes a monophasic sleep at night with periodic slow wave sleep (SWS)

and REM sleep alternation. Furthermore, electrophysiological experiments and pharmacological treatment canbe applied without much difficulty as in rodents. It may be suitable for sleep research, however, the basic
properties of sleep inremain elusive. In this study, we examined whether sleep homeostasis is observedin response to sleep deprivation (SD). 7 hours of SD by light and gentle handling at night showed delayed wakeup time compared with a control group, suggesting the presence of sleep homeostasis in. Furthermore,

LFP analysis showed that SD affected the periodic oscillation of SWS and REM sleep, as in humans.
These results indicate that the features of sleep, especially sleep homeostasis in each sleep stage are similar
between reptiles and mammals. Observation/manipulation of reptilian sleep may open a new avenue forunderstanding the fundamental functions and circuit mechanisms of monophasic sleep.

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学生セッション

Construction of minimal neural circuits by iPSC-derived astrocytes for glial drug
discovery〇内野 鉱也1、田中 泰圭2,3,4、川口 紗果1、窪田 香織1、渡辺 拓也1、桂林 秀太郎1,4、廣瀬 伸一3,4,5、岩崎 克典11

福岡大・薬・臨床疾患薬理学教室、2第一薬科大・薬・薬物治療学分野、3株式会社iONtarget、4福岡大・てんかん分子研究所、5福岡大・医・総合医学研究センター

Astrocyte is a type of glial cell involved in synaptic transmission and the formation and maturation of synapses.
Currently, the establishment of induced pluripotent stem cells (iPSCs) allows the differentiation of stem cells
into various types of cells while preserving the patient phenotype. Therefore, patient iPSCs replace animalmodels in pathological analysis and drug discovery. Technological advances have provided access to human
iPSC-derived astrocytes (HiAs). Furthermore, neurons co-cultured with pathological astrocytes have been used
to study their morphology, protein levels, and spontaneous synaptic responses. However, these studies did notinvestigate detailed synaptic functions such as synaptic transmission evoked by electrical stimulation andmorphological analysis at the single neuron level. In this study, we established autaptic cultures with HiAs (HiAs
Autaptic Cultures, HiAACs), single neuron cultures grown in isolation on microislands of HiAs that form
synapses exclusively with themselves. We found that neurons in HiAACs develop morphologically by co-culture
with HiAs and form functional synapses that exhibit excitatory postsynaptic currents. Although we used healthy
astrocytes in this study, HiAACs can be used to study various diseases by using patient-derived astrocytes in the
future.This work was supported by funding from JSPS, AMED, MEXT, the Science Research Promotion Fundand The Fukuoka University Fund, and Kyushu University Hospital.

Copyright © The Japanese Pharmacological Society3-B-SS13-5

学生セッション

Cellulose rich food induces intestinal disturbance, anxiety-like behavior andamygdalar dopaminergic hyperactivity in mice.

〇伊東 楓1、原口 敦嗣1、野崎 千尋1,2、柴田 重信1
1

早稲田大・先進理工・電気・情報生命、2早稲田大・国際理工学センター・Major in bioscience

It is indicated that the intestinal environment affects the brain, which is known as the gut-brain axis. In fact,
improvement of intestinal environment is reported to suppress anxiety-like behavior in mice model ofdepression and schizophrenia. However, its detailed mechanisms remain unclear, particularly in context ofintestinal environment effects on emotion in non-disease mice. The object of this experiment is therefore toverify the effect of intestinal environment on anxiety-like behavior and its detailed physiological mechanisms.
We previously found that cellulose rich food (AIN-93M) suppresses the production of short-chain fatty acid andexacerbate the intestinal condition. In the present study, we fed mice with AIN-93M to modify the intestinal
environment, or the mouse chow that can maintain a favorable intestinal environment (MF). After 8-weeks
feeding, AIN-93M-fed animals displayed the significant increase of marble-burying behavior compared to MFfed group, suggesting that dysfunction of intestine lead by AIN-93M enhanced anxiety-like behavior.
Furthermore, dopamine release as well as dopamine receptor expression has been increased in amygdala of AIN-93M fed mice but not in MF animals, indicating that enhanced anxiety-like behavior in AIN-93M animals isdue to such modification of amygdalar dopaminergic system.
These results suggest that cellulose rich food may exacerbate intestinal environment, which may enhance theanxiety-like behavior and overactivation of dopaminergic system.

Copyright © The Japanese Pharmacological Society3-B-SS13-6

学生セッション

Circulating apolipoprotein B-100 promotes scar formation after spinal cord injury〇米津 好乃1,2、田辺 章悟1、三澤 日出巳2、村松 里衣子1
国立精神・神経医療研究セ・神経研・神経薬理、2慶應義塾大・院薬・薬理学

1

Spinal cord injury (SCI) causes severe neurological dysfunction depending on the neuronal network disruption.
Neuronal network regeneration is prevented by the scar tissue formed around the lesion; therefore, inhibition ofscar formation is considered to contribute to neuronal network regeneration, resulting in functional recovery. It
is known that the scar is composed of several cell types. Among them, recent studies indicated that pericytes act
as key players in scarring triggered by their proliferation. Thus, understanding the molecular mechanism
regulating pericyte proliferation may be useful to promote neuronal network regeneration after SCI; however,
the mechanism is not fully elucidated. Here we focused on the disruption of vascular barrier, a histologicalfeature around the lesion, and found that circulating apolipoprotein B-100 (ApoB-100) promotes pericyte
proliferation which contributes to the scar formation after SCI. CRISPR-Cas9 knockout screens with primarymouse pericytes identified that ApoB-100/low-density lipoprotein receptor (LDLR) signal promotes pericyte
proliferation. ApoB-100 knockout mice exhibited a decrease in pericyte-derived scar formation and a significant
improvement of motor function after SCI. These results suggest that circulating factors, especially ApoB-100,
could be a novel therapeutic target for treating SCI.

Copyright © The Japanese Pharmacological Society3-B-SS14-1

学生セッションChronic social stress-induced transition of microglial transcriptome states inmice〇三島 零、谷口 将之、松下 和敏、古屋敷 智之
神戸大・院医・薬理

Microglia-derived neuroinflammation has been associated with the stress pathology of mental illness. However,the nature of microglial stress responses remains poorly understood. Using single-cell transcriptome analyses,
we found that acute and chronic social defeat stress altered the transcriptome of microglia in multiple brain
areas, including the medial prefrontal cortex (mPFC), primary motor and sensory cortices, hippocampus,
nucleus accumbens, and hypothalamus, in mice. Despite some brain region-specific patterns, individual
variability of stress-susceptibility emerged broadly across the brain areas. We further analyzed transcriptomes ofmPFC microglia, the activation of which are essential for chronic stress-induced depression-like behavior, andidentified several transcriptomic states of microglia, through which chronic stress promoted the transition froma homeostatic state to a distinct state with brain region-specificity and stress susceptibility signature. Thesefindings demonstrate multiple states of microglia reflecting brain area specification and stress susceptibility, thetransition of which may contribute to the stress pathology of mental illness.

Copyright © The Japanese Pharmacological Society3-B-SS14-2

学生セッションThe Use of an Amyloid beta Intracerebroventricular Model to Investigate theInteraction between Alzheimer‘s Disease and Circadian Dysfunction〇ラベル リンダ1、野崎 千尋2、田原 優3、柴田 重信1
1

早稲田大・理工・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、3広島大学大学院・医系科学研究科・公衆衛生学

There are 40 million people worldwide living with Alzheimerʻs Disease (AD), and by 2050 the prevalence is
expected to increase to 150 million people, thus resulting in a 1 in 2 chance of having AD by the age of 85. An
increase in amyloid-b plaques (Ab) and hyperphosphorylated tau are widely accepted as the core indicators ofthe disease. These indicators of AD onset and progression are becoming increasingly attributed to the
disturbance of optimal sleep/wake cycles. Additionally, AD progression exacerbates normal sleep/wake cycles,
resulting in a cyclical worsening of AD pathology, circadian rhythm, and cognitive dysfunction. By using an Ab
intracerebroventricular (ICV) injection mouse model, mice can present an AD-like pathology within days toweeks after injection, allowing for the expedited examination of the disease. This model can serve as a tool toinvestigate the mechanism behind the disturbance of the biological clock and AD progression, and therefore
find therapeutic targets to delay, prevent or cure the disease.

Copyright © The Japanese Pharmacological Society3-B-SS14-3

学生セッション

Food-derived hydrophilic amino acid ergothioneine prevents cognitive decline in
Alzheimer‘s disease model mice at its clinically relevant plasma concentrations.

〇松尾 雄介1、石本 尚大1、増尾 友佑1、笹栗 弘貴2、齊藤 貴志2、西道 隆臣2、加藤 将夫1
金沢大・薬学系、2理研・脳神経科学研究セ

1

Progressive pathology of Alzheimer's disease (AD) including aggregation of amyloid beta (Aβ) appears decades
before the onset of dementia. Therefore, early intervention before the appearance of the symptoms is crucially
important. We focused on food-derived hydrophilic amino acid ergothioneine (ERGO) because oral ERGO
administration enhances cognitive function in normal mice. The purpose of the present study is to elucidate thepreventive effect of ERGO on cognitive decline in the AD model,

mice expressing humanized Aβ.

mice were orally administered ERGO or vehicle weekly between 5 weeks to 7 months of age. Novelobject and spatial recognition tests showed that ERGO administration significantly improved the cognitive
declines in themice. ERGO concentration in plasma ofmice at 3 months of age reached asteady state around 10 µM, which was close to that reported in humans orally administered ERGO. Proteome
analysis of the hippocampus samples showed that 71 and 91 proteins were significantly up- and down-regulated
more than two-fold by ERGO treatment, respectively. Further enrichment analysis revealed that neurogenesisrelated proteins were significantly enriched. Immunochemical analysis in the hippocampus showed that area ofnewborn neuron marker doublecortin-positive cells in the ERGO-treatedmice was significantly higher

than that in vehicle-treated mice. These results suggest that ERGO would prevent cognitive decline at least
partially via the promotion of neurogenesis inmice.

Copyright © The Japanese Pharmacological Society3-B-SS14-4

学生セッション

Synaptotagmin 4 contributes to spontaneous regeneration of neural networks
after spinal cord injury〇樋口 京香1,2,3、田辺 章悟1、成田 年3,4、村松 里衣子11

国立精神・神経医療研究セ・神経薬理研究部、2東京医科歯科大・院医歯・NCNP脳機能病態学、3星薬科大・薬・薬理、4国立がん研・がん病態生理

Spinal cord injury (SCI) causes severe neurological dysfunction such as motor deficits and sensory impairments
caused by the disruption of neuronal networks. Injured neurons in the central nervous system spontaneously
regenerate their neuronal network, resulting in promoting functional recovery. However, the regenerative
capacity in the adult central nervous system is limited, therefore, understanding the mechanism that promotes
regeneration of the neuronal network is required to establish a strategy for restoring motor function after SCI.
In this study, we explored the factor that promotes neurite elongation using
that(siRNA-screening and foundhas a potential to promote neurite elongation. We detected that

expressed in neurons in the brain compared with other organs. To ask the function of
adeno-associated virus 9 (AAV9) encoding shRNA against
suppression ofis mainly

in vivo, we injected

into the motor cortex and found thatprevented regeneration of neuronal network and functional recovery in the mice with SCI.

These results indicate that

expressing in neurons sustains the spontaneous regeneration of the neuronal

network in the adult central nervous system.

Copyright © The Japanese Pharmacological Society3-B-SS14-5

学生セッション

Interaction of Munc13-1 and RIM that contributes to the formation/maintenanceof synaptic vesicle release sites〇小島 佑介、坂本 寛和、並木 繁行、廣瀬 謙造
東京大・院医・細胞分子薬理学

Neurotransmitter release is regulated by several proteins localized at the active zones of presynaptic terminals.
Munc13-1 is a multi-domain active zone protein which redundantly interacts with other active zone proteins
including RIM. We have previously shown that Munc13-1 forms supramolecular nanoassemblies that function
as the synaptic vesicle release sites. Here, we aimed to clarify a more detailed molecular mechanism of theformation/maintenance of Munc13-1 nanoassemblies by intervening in the interaction of Munc13-1 with RIM.
For this purpose, we isolated the Zn2+ finger domain of RIM (RIM-ZF), which interacts with the C2A domain of
Munc13-1 and expressed it in neurons to competitively inhibit the binding of endogenous Munc13-1 and RIM.
Glutamate imaging revealed that the expression of RIM-ZF in cultured neurons caused a significant decrease inneurotransmitter release, although a previous study showed that the RIM-ZF partially rescues suppressed
neurotransmitter release in RIM knock-out neurons. Furthermore, quantitative immunocytochemical analysis
with super-resolution microscopy revealed that the expression of RIM-ZF altered the nanoscale distribution of
Munc13-1 molecules at the active zones. Thus, our results suggest that a direct interaction of RIM-ZF with
Munc13-1 itself is incomplete for the appropriate formation of synaptic vesicle release sites, and that crosslinkage of RIM-ZF to the other domains of RIM is necessary for the precise positioning of Munc13-1 at theactive zone.

Copyright © The Japanese Pharmacological Society3-B-SS14-6

学生セッション

Skeletal muscle atrophy-induced hemopexin accelerates cognitive dysfunction
in 5XFAD mice

〇井城 綸沙、東田 千尋
富山大・和漢医薬学総合研究所・神経機能学領域

Physical inactivity is one of risk factors for Alzheimerʻs disease (AD). Performing physical exercise is difficult at
old age, and thus, decline in physical movement may be a cause of age-associated lowering of the brain function.
This study aimed to elucidate the onset of the skeletal muscle atrophy-induced acceleration of AD and its
molecular mechanism.
Presymptomatic AD model (5XFAD) mice were used. The bilateral hindlimbs were immobilized by castattachment for 14 days. Wet weight of hindlimbs muscles were significantly lower in cast-attached 5XFAD micethan those in non-cast mice. At the same time, object recognition memory in the cast-attached 5XFAD mice was
impaired. The hindlimb muscles were organ cultured. And the conditioned media (CM) was separated by 2DPAGE and analyzed by MALDI-TOF MS. The most increased spot in the cast-attached muscle CM washemopexin. Hemopexin levels in the skeletal muscle, plasma, and hippocampus were increased in cast-attached
5XFAD mice. Continuous i.c.v. infusion of hemopexin for 2 weeks induced memory deficits in young 5XFAD
mice. Gene microarray analysis of the hippocampus was performed to investigate the molecules involved inmemory deficit. Lipocalin-2 (Lcn2) mRNA, neuroinflammation-associated factor, was increased in the
hippocampus in hemopexin-infused 5XFAD mice compared to in control mice.
These findings provide new evidence indicating that skeletal muscle atrophy has an unbeneficial impact on theoccurrence of memory impairment in 5XFAD mice, which is mediated by skeletal muscle atrophy-driven
hemopexin.

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