bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adaptive cytoprotection and cancer chemoprotection by sulforaphane against various stresses: role of nrf2‐keap1 dependent and independent pathways (1052.1)
Authors: Yanaka Akinori 
Published: June 2021

Background and Aims: Mild irritants protect GI mucosa from more severe stress, called “adaptive cytoprotection”. Prostaglandins have been proposed as mediators for this phenomenon. In contrast, recent studies show that nrf2‐dependendent anti‐oxidant system may be responsible for this protection, since most of the physiological stresses are oxidative stress in GI tract. In this study, we examined if sulforaphane (SFN), a nrf2‐stimulating phytochemical rich in broccoli sprouts, protects GI mucosa against various physiological stresses. We also examined if SFN inhibits tumor formation in chemically induced colon cancer model in mice.Methods: 1. Adaptive Cytoprotection by SFN: 1)In vitro model for intestinal injury: IEC6 cells were exposed to aspirin with or without SFN. 2) In vivo model for indomethacin‐induced small intestinal ulcer. Mice were treated with indomethacin (IND) with or without SFN. 2. Chemoprotection by SFN: C57/BL6 mice, pretreated with a chemical carcinogen, azoxymethane, were fed for 8 weeks with or without sulforaphane glucosinolate (SGS). Number of aberrant crypt foci (ACF), a precancerous lesion, in colonic mucosa in each mice were counted.Results: 1. 1) Pretreatment with SFN up‐regulated antioxidant enzymes activity, and protected IEC cells from aspirin‐induced injury. 2) Dietary SGS protected small intestine from IND‐induced injury. These effects were mitigated by a HO‐1 inhibitor, and were abolished in nrf2‐gene knockout mice.2. SGS suppressed formation of ACF in azoxymethane‐treated mice, effects accompanied by inhibition of the cell proliferation, and enhancement of the apoptosis. These effects were demonstrated in both nrf2+/+ and nrf2‐/‐ mice.Conclusions: 1. SFN affords adaptive cytoprotection of small intestine against NSAIDs‐induced oxidative stress via induction of the nrf2‐dependent anti‐oxidant enzymes. 2. SFN affords chemoprotection against colon cancer via nrf2‐independent mechanisms.
Sevoflurane enhances proliferation of human colon cancer HCT116 cells via adenosine triphosphate‐sensitive potassium channels (1052.8)
Authors: Kawaraguchi Yoshitaka , Sugimoto Hiroshi , Nomura Yasumitsu , Furuya Hitoshi , Kawaguchi Masahiko 
Published: June 2021

Introduction: Sevoflurane is one of the commonly used volatile anesthetics in cancer patients. The protective effect of sevoflurane preconditioning against ischemia/reperfusion has raised concerns about whether sevoflurane could acts advantageously for survival even in cancer cells. Therefore, we investigated the effects of sevoflurane on proliferation in cancer cells.Methods: HCT116 cells were plated in 96‐well plates at a density of 1 × 104 cells/well and incubated overnight. On the following day, cells were exposed to 1% sevoflurane for 6 h. After 24 h recovery, we performed MTT assay. The absorbance of the formazan product was measured at a wavelength of 570 nm using 650 nm as the reference. In addition, to investigate the role of adenosine triphosphate‐sensitive potassium (KATP) channels, we conducted the same experiment under co‐administration with KATP inhibitor, glibenclamide.Results: Sevoflurane significantly enhanced cell proliferation compared to control (0.53+0.023 vs. 0.46+0.033, n=6, p<0.01, respectively). Enhanced proliferation by sevoflurane was completely blocked by co‐administration with glibenclamide.Conclusion: We show that 1% sevoflurane for 6 h enhanced cell proliferation via KATP channels in HCT116 cells.
A study of the effects of 25wssw on anti‐endotoxemia (1056.4)
Authors: Yun Sun , Ya Yuan , Yao Ren , Teng Zhang , Jie Li , Hua Wang , Qin Zhao 
Published: June 2021

OBJECTIVES: The aim of this study is to investigate the anti‐endotoxemia effect of 25wssw. METHODS: 25WSSW was dissolved in water. Then collected the supernatant. 24 SD rats(200g‐240g)were divided into four groups randomly: NS group, high 25wssw dosage group(HDG), low 25wssw dosage group(LDG), dexamethasone group (DXM). DXM Intraperitoneal injection(2mg/Kg) for dxm group. 25wssw Introgastric administration 1 time per day for 5 days to HDG and LDG. Then each group was invaded by endotoxin. 3h after, venous blood was collected, and the endotoxin concentration in serum was tested by Endotoxin tester. NEUT and WBC concentration were tested by automatic biochemistry analyzer. RESULTS: 3h after endotoxin injection, compared with NS group, the endotoxin concentration in serum at HDG and LDG was deseased(p<0.05), the endotoxin concentration in serum at DXM group was deseased either,but compared with NS group, P>0.05. And the amount of WBC at DXM group was obviously increased, but the amount of WBC at HDG tended to be decreased. There’s no significant difference to the amount of NEUT . CONCLUSION: 25wssw has the anti‐endotoxemia effects by neutralizing endotoxin in serum, and by decreasing the process of inflammation.Grant Funding Source: Supported by 2011 Natural Science Fund Project of Tibet science and technology department
Vasorelaxant response of coronary arteries to capsaicin and effect of angiotensin II (1057.1)
Authors: Yang Qin , Pun Shun‐Hay , Yu Cheuk‐Man 
Published: June 2021

Vasorelaxant effects and mechanisms of the TRPV1 selective agonist capsaisin and the effect of angiotensin II on capsaicin‐induced relaxation were studied in small porcine coronary arteries (PCAs). In U46619‐constricted PCAs, capsaisin induced dose‐dependent relaxation with significant response starting at 0.1µmol/L and full relaxation at 100µmol/L. Endothelium denudation significantly inhibited relaxant responses to capsaicin at concentrations of 0.1‐10µmol/L. sPCAs pretreated with IKCa and SKCa channel blockers, TRAM‐34 and apamin, showed suppressed relaxation to capsaicin (0.1‐10µmol/L) and the inhibition was comparable to that in PCAs constricted with 40mmol/L KCl (35.5±4.7%, 29.5±3.9% vs. 79.0±4.3% in control, p<0.001). However, relaxation to 100µmol/L capsaicin remained unaffected by endothelium denudation, IKCa and SKCa blockade, or high [K+]o. Exposure of sPCAs to angiotensin II (100nmol/L) for 24h inhibited capsaicin‐induced relaxation (33.3±3.5%, p<0.001). Capsaisin induces relaxation of PCAs through both endothelium‐dependent and ‐independent mechanisms. IKCa and SKCa are involved in the endothelium‐dependent mechanism. Angiotensin II impairs capsaicin‐induced endothelium‐dependent relaxation.Supported by GRF CUHK4774/12M; NSFC 81200123; CUHK Direct Grant 4054015; Key Med. Program of Tianjin Binhai Health Bureau 2011BHKZ001; Tianjin Health Bureau 2012KZ009.Grant Funding Source: Supported by GRF CUHK4774/12M; NSFC 81200123; CUHK Direct Grant 4054015
Pharmacological dissection reveals functional roles for KCNQ channel subtypes in human urinary bladder smooth muscle contractility (1057.12)
Authors: Provence Aaron , Rovner Eric , Petkov Georgi 
Published: June 2021

This study is aimed at investigating the molecular expression of voltage‐gated potassium (KCNQ) channels in urinary bladder smooth muscle (UBSM) and to reveal their functional role in regulating the contractility of this tissue. Using native, freshly‐isolated human UBSM single cells and whole UBSM tissue, we examined the mRNA and protein expression for all KCNQ channel subtypes and elucidated their functional role in human UBSM contractility using RT‐PCR, western blot, and isometric UBSM tension recordings of isolated human UBSM strips. Human UBSM tissues were collected from open bladder surgeries and represent patients without preoperative symptoms of overactive bladder. Expression of KCNQ channel subtypes were identified at both the mRNA and protein levels in human UBSM. Functional studies revealed that spontaneous phasic contractility was significantly increased by the KCNQ channel inhibitor, XE991. On the contrary, the KCNQ channel activators retigabine, L‐364,373 (R‐L3), and ICA‐069673 attenuated UBSM spontaneous phasic contractions. This study demonstrates the novel findings that KCNQ channels are functionally expressed in human UBSM, whereby they serve an important functional role in regulating the spontaneous contractility of this tissue.Grant Funding Source: Supported by NIH R01 DK084284 to Georgi V. Petkov
Significance of store operated calcium entry in human abdominal aortic aneurysm vascular smooth muscle cells (1057.3)
Authors: Bailey Marc , Young Richard , Rode Baptiste , Foster Richard , Li Jing , Beech David 
Published: June 2021

Introduction Abdominal aortic aneurysm (AAA) is a focal dilatation of the abdominal aorta associated with defective vascular smooth muscle cell (VSMC) function. Store operated calcium entry (SOCE) through Orai1 can modulate cell behaviour. We investigate if SOCE is present in AAA VSMC and if pharmacological blockade impacts cellular behaviour.Methods VMSC were cultured from human AAA and internal mammary artery (IMA). Gene expression was studied by real‐time PCR, calcium imaging using fura‐2AM in response to thapsigargin or platelet derived growth factor (PDGF) and functional assays (proliferation, migration, apoptosis) by time‐lapse fluorescent microscopy. The SOCE inhibitor S66 and its novel analogue JPIII were compared to vehicle control (dimethylsulfoxide).Student t‐test was used to compare responses.Results Orai1/STIM1 mRNA was detected in both cell populations and a SOCE response observed, which was inhibited by S66 or JPIII. The proportion of signal blocked was greater in the AAA VSMC than the IMA VSMC after PDGF stimulation. In functional experiments, treatment of the AAA VSMC with JPIII had no effect on proliferation or migration but conferred protection against apoptosis, which was itself increased in the AAA compared to the IMA VSMC.Conclusion SOCE is functional in AAA VSMC and inhibition protects against apoptosis. The channels warrant further study as a potential therapeutic avenue for AAA.Grant Funding Source: Supported by the British Heart Foundation
Impact of TRPC channels on body weight (1057.9)
Authors: Rode Baptiste , Yuldasheva Nadira , Wheatcroft Stephen , Ainscough Justin , Beech David 
Published: June 2021

We previously showed that Transient Receptor Potential Canonical (TRPC) channels are expressed by adipocytes and that short‐term inhibition of these channels increased the concentration of circulating adiponectin, suggesting a functional role of TRPC channels in adipocyte biology in vivo (Sukumar et al, Circ Res. 2012;111:191‐200). The aim of this study was to investigate if there are implications of TRPC channels for the control of body weight. Mice with conditional global expression of dominant‐negative ion pore mutant TRPC5 were used to suppress ion permeation in TRPC channels in vivo in adults without deleting TRPC proteins. Mice were fed a western‐style high fat diet for 12 weeks and male litter‐mates were compared with and without suppression of TRPC ion permeation. Mice expressing mutant TRPC5 showed significantly lower body weight as compared to controls. No obvious adverse effects on health were evident. Analysis of adipose tissue showed that adiponectin mRNA was more abundant in the mice expressing mutant TRPC5, suggesting regulation of gene expression in adipocytes. The results suggest that ion flux through TRPC channels is a driver for weight gain and that ion pore blockers of TRPC channels could be an approach for protecting against obesity.Grant Funding Source: Supported by the Medical Research Council
Subpopulation of highly reactive cysteines underlie the activation of TRPA1 by electrophiles (1061.6)
Authors: Taylor‐Clark Thomas , Bahia Parmvir 
Published: June 2021

Transient Receptor Potential Ankyrin 1 is a cation channel expressed on nociceptive sensory nerves. TRPA1 is activated by electrophiles following covalent modification (Michael Addition), and this mechanism contributes to nociceptor activation (and pain/defensive reflexes) during oxidative stress. However, the rate of electrophile‐induced TRPA1 currents (~1000 M‐1s‐1) is much faster than the electrophilic covalent modification of canonical Cys residues (<3 M‐1s‐1). The basis for this high reactivity is not known. Using overexpression of V5‐tagged TRP channels and pulse‐chase experiments with fluorescent electrophile BODIPY‐iodoacetamide (BD‐IA), we have studied TRP channel covalent modification following immunoprecipitation. Human TRPA1 is rapidly bound by BD‐IA (<60s, 10uM), with ~50% of the binding sensitive to a short pretreatment with IA. Other TRP channels are slowly bound by BD‐IA, but this binding is not blocked by IA pretreatment. As Michael Addition requires ionization of thiols we determined the pKa of TRP channel Cys. The IA‐blockable Cys residues of human TRPA1 had very low pKa (~3) compared to the IA‐insensitive Cys residues (>8). In addition, TRPA1 binding of electrophiles is not modified by intracellular polyphosphates (required for TRPA1 currents) or TRPA1 antagonists. We conclude that a subpopulation of reactive Cys is responsible for TRPA1’s unusual activation mechanism.Grant Funding Source: Supported by R01HL119802‐01
Pharmacokinetics and metabolism of clausenamide enantiomers (1064.4)
Authors: Zhu Chuan‐jiang , Zhang Jun‐tian 
Published: June 2021

The properties of (‐) and (+)clausenamide(CLA) in pharmacokinetics and metabolism were investigated in rats and rabbits. There were several main findings obtained from these studies: (1) (+)CLA exhibited greater tmax(35.0±12.2 min, p<0.01), Cmax (26.1±7.1μg/ml), t1/2β(217.2±34.2min, p<0.01), AUC0‐12h (2446.0±540.4 min/μg/ml, p<0.001), and smaller CL/F(0.063±0.017 l/kg/min, p<0.001) and Vd/F(20.0±7.8 l/kg, p<0.01), than (‐)CLA after oral administration to rats. The (+)/(‐)‐isomer ratios for tmax, Cmax, t1/2β, AUC0‐12 h, CL/F, and Vd/F were 1.75, 1.21, 1.26, 1.95, 0.47, and 0.59, respectively; (2) the binding of (+)CLA to rat plasma protein (38.0%±2.6%) was significantly greater than that of (‐)CLA (28.5 %±0.6%); and there were significant species differences in the plasma protein binding of (‐)CLA between rabbits(47.2 %±4.9 %) and rats. (3) (‐)CLA had a more rapid absorption and distribution to rat hippocampus, cortex, and cerebellum with smaller values for tmax, t1/2β, and AUC0→∞, and greater ones for CL/F and Vd/F than its antipode. (4) (‐)CLA was mainly excreted in feces with 13.9% of dose, whereas (+)CLA in bile with 17.2%; (5) in the in vitro metabolic system, (‐)CLA was mainly metabolized to 7‐hydroxy‐, 5‐hydoxy‐ and 4‐hydroxy‐CLA, and 7‐hydroxylation was a preferential pathway , but (+)CLA was mainly metabolized to 4‐hydroxy‐CLA, its 7‐hydroxyl and 4‐hydroxyl metabolites were very small; (6)the major metabolic enzyme for CLA enantiomers was CYP3A isoforms whose expression could be induced by (‐)‐isomer in the rat liver. (7) both (‐) and (+)CLA were subjected to an intermediate degree of first‐pass metabolism in rabbits with rates of hepatic extraction 64.7% for (‐)‐isomer and 50.8% for (+)‐isomer. These data suggested that there were markedly stereoselective differences in pharmacokinetics and metabolism between two enantiomers, which may be important in understanding the clinic pharmacology of active eutomer, (‐)CLA, for treatment of Alzheimer’s disease.Grant Funding Source: grants from the National Natural Science Foundation of China (29790120; 30873117).
GPCRs as potential therapeutic targets in pancreatic cancer‐associated fibroblasts (1066.11)
Authors: Zhou Shu , McCann Thalia 
Published: June 2021

GPCRs as potential therapeutic targets in pancreatic cancer‐associated fibroblasts
Shu Zhou1, Thalia McCann1, Randall French2, Andrew M. Lowy2,3, Paul A. Insel1,4 1Department of Pharmacology, 2Moores Cancer Center, 3Division of Surgical Oncology, Department of Surgery, 4Department of Medicine, UCSD, La Jolla, California Pancreatic ductal adenocarcinoma (PDAC) is characterized by a dense fibrotic stromal matrix, composed of activated fibroblasts/stellate cells, inflammatory cells and other cell types. This unique tumor microenvironment has been increasingly recognized as a key mediator of PDAC progression and drug resistance. Thus, targeting the tumor stroma may be a novel therapeutic approach for PDAC. We hypothesized that G‐protein‐coupled receptors (GPCRs) expressed by pancreatic cancer‐associated fibroblasts (CAFs) may be potential therapeutic targets for PDAC. To begin to test this hypothesis, we used an unbiased GPCRomic array approach to identify and quantify the GPCRs expressed by pancreatic CAFs. We found 125 GPCRs, whose expression was shared in CAFs obtained from the primary tumors of five patients. Among those GPCRs, 35 had at least two‐fold higher expression in the CAFs compared to normal pancreatic fibroblasts and included GPCRs that link to each of the major classes of G proteins. RT‐PCR analysis of the 10 most up‐regulated GPCRs correlated well (r2=0.88) with the data from the GPCR array. We conclude that: 1) GPCRomic array analysis can identify and quantify the profile of GPCRs expressed by pancreatic CAFs; 2) the CAF‐expressed profile differs from that of normal pancreatic fibroblasts and 3) GPCRs expressed by CAFs may have functional roles and be novel therapeutic targets for PDAC.Grant Funding Source: Supported by Cancer Therapeutics Training Program
Angiotensin‐II mediates ACE2 internalization and degradation through an angiotensin‐II type I receptor‐dependent mechanism (1066.9)
Authors: Filipeanu Catalin , Deshotels Matthew , Xia Huijing , Lazartigues Eric 
Published: June 2021

ACE2 mediates the conversion of ang‐II to ang‐(1‐7) and by this mechanism prevents the pathological effects related to the activation of rennin‐angiotensin system (RAS). We previously reported that decreased ACE2 expression and activity leads to the development of Ang‐II‐mediated hypertension in mice. The aim of the present study was to characterize the mechanisms involved in ACE2 down‐regulation during neurogenic hypertension. In ACE2‐transfected Neuro‐2A cells, Ang‐II treatment resulted in a significant attenuation of the enzyme activity. Confocal microscopy experiments demonstrated that in presence of ang‐II, ACE2 is internalized and co‐localizes with Rab7, a lysosomal marker. These effects were prevented by the AT1R blocker losartan and the lysosomal inhibitor leupeptin. In contrast, in HEK293T cells, which lack endogenous AT1R, Ang‐II failed to promote ACE2 internalization. This effect was restored after AT1R transfection. Further, ang‐II enhanced ACE2 ubiquitination and decreased the interactions between AT1R and ACE2. In contrast, ACE2 activity was not changed by transfection of AT2R or Mas1R. In vivo, Ang‐II‐mediated hypertension was blunted by chronic infusion of leupeptin in C57Bl/6, but not in ACE2 knockout mice. This is the first demonstration that elevated Ang‐II levels reduce ACE2 expression and activity by stimulation of lysosomal degradation through an AT1R‐dependent mechanism.Grant Funding Source: P20‐RR‐018766, RO1‐HL093178, Howard University
NHERF‐1 expression in aging kidney tubular apical and basolateral membranes (1067.11)
Authors: Khundmiri Syed , Murray Rebecca , LeBlanc Amanda , Sen Utpal , Lederer Eleanor , Jose Pedro , Weiman Edward 
Published: June 2021

NHERF‐1 regulates several ion transporters in kidney proximal tubules including NHE3, NpT2a, NBC, CFTR, and Na‐K ATPase (NKA) through modulation of protein complex formation and through modulation of hormone receptor signaling, in particular, PTH and dopamine. Regulation of ion balance changes with aging; however, the expression of NHERF‐1 in aging has not been studied. We hypothesize that loss of NHERF‐1 regulation of NHE3 and NKA with aging contributes to alterations in maintenance of sodium balance. To address this hypothesis, we compared expression of NHERF‐1, NHE3, and NKA in kidneys from young (4 m old) and old (24 m old) female F344 rats by western blot in isolated apical and basolateral membranes and by immunohistochemistry. Western blot showed complete loss of NHERF‐1 in both apical and basolateral membranes from kidneys of 24 m old rats. NKA expression significantly decreased in 24 m old rats as compared to 4 m old rats, but NHE3 expression did not change with aging. Similar to aging, NHERF‐1 expression was also found to be absent in proximal tubule cells from spontaneous hypertensive rats. These data suggest that in SHR and aging dependent alterations in ion homeostasis may result from changes in transporter expression and from loss of normal hormonal regulatory pathways. Supported by AHA and VAGrant Funding Source: AHA, VA
The effect of peroxisome proliferator activated receptor ‐α activation and apocynin on superoxide dismutase and NADPH oxidase expression in the brains of mice during angiotensin II‐induced hypertension (1067.9)
Authors: Bouchelion Michelle , Mervin Addias , Lee Dexter , Allard Joanne 
Published: June 2021

Approximately 33% of American adults suffer from hypertension which significantly increasing their risk for heart attacks and strokes. Hypertension is also associated with elevated levels of reactive oxygen species (ROS), which can negatively affect cellular function. Studies suggest that NADPH oxidase enzymes (NOX) are major sources of ROS in both the central nervous systems and that NOX levels are increased with hypertension. Superoxide dismutase (SOD), a powerful antioxidant and anti‐inflammatory protein acts as a scavenger of ROS. Administration of compounds that increase SOD levels can provide protection from the harmful effects of hypertension. This study’s objective is to determine the effects of two anti‐oxidative drugs, fenofibrate, a peroxisome proliferator activated receptor‐α agonist, and apocynin, a NADPH oxidase inhibitor, on brain levels of NOX and SOD, in mice with Angiotensin II (Ang II) ‐induced hypertension. We hypothesize that Fenofibrate (145 mg/kg/day) and Apocynin (1 g/L) will attenuate the effects of Ang II by increasing SOD levels and decreasing the levels of NOX enzymes. Male Swiss‐Webster mice were divided into 4 groups. Control (C) mice were untreated. All other mice received Ang II (400 ng/kg/min) for 12 days. Apocynin and Fenofibrate groups received additional doses of either fenofibrate (145 mgkg/day) or Apocynin (1 g/L). Mean arterial pressure on day 12 of Ang II was 100 ± 3, 120 ± 1, 112 ± 5, 116 ± 1 mmHg for Control, Ang II, Ang II + Fenofibrate and Ang II + Apocynin, respectively. Mice were sacrificed, brains were dissected, and protein levels were evaluated using standard western blotting procedures. Results suggest that fenofibrate decreases SOD levels while increasing NOX levels in brain. These preliminary results do not support our initial hypothesis; therefore additional experiments will be conducted to elucidate the mechanisms of fenofibrate and apocynin in this mouse model of hypertension.Grant Funding Source: NHLBI 5K01HL092593‐05