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Role of arachidonic acid in hyposmotic membrane stretch-induced increase in calcium-activated potassium currents in gastric myocytes

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Acta Pharmacologica Sinica 2005年第10期26卷 1233-1242页

作者： Meng YANG~2, Xing-lan LI~3, Hui-ying XU~3, Jia-bin SUN~3, Bin MEI~4, Hai-feng ZHENG~3, Lian-hua PIAO~3, De-gang XING~3 Zhai-liu LI~3, Wen-xie XU~(2,5) ~2 department of physiology, Shanghai Jiaotong university School of medicine, Shanghai 200030 ~3.department of physiology yanbian university college of medicine yanji 13.000 ~4 East China Normal university School of Life Science, Shanghai 200062, China departmentofphysiology ShanghaiJiaotongUniversitySchoolofMedicineShanghai200030 departmentofphysiology YanbianUniversityCollegeofMedicineYanji133000 EastChinaNormaluniversitySchoolofLifeScience Shanghai200062

Aim: To study effects of arachidonic acid （AA） and its metabolites on the hyposmotic membrane stretch-induced increase in calcium-activated potassium currents (IKCa) in gastric myocytes. Methods: Membrane currents were recorded by using a conventional whole cell patch-clamp technique in gastric myocytes isolated with collagenase. Results: Hyposmotic membrane stretch and AA increased both IK（Ca）and spontaneous transient outward currents significantly. Exogenous AA could potentiate the hyposmotic membrane stretch-induced increase in IK（Ca）The hyposmotic membrane stretch-induced increase inK（Ca）was significantly suppressed by dimethyleicosadienoic acid （100 μmol/L in pipette solution）, an inhibitor of phospholipase A2. Nordihydroguaiaretic acid, a lipoxygenase inhibitor, significantly suppressed AA and hyposmotic membrane stretch-induced increases in IK（Ca）. External calcium-free or gadolinium chloride, a blocker of stretch-activated channels, blocked the AA-induced increase in IK（Ca）significantly, but it was not blocked by nicardipine, an L-type calcium channel blocker. Ryanodine, a calcium-induced calcium release agonist, completely blocked the AA-induced increase inK（Ca）; however, heparin, a potent inhibitor of inositol triphosphate receptor, did not block the AA-induced increase in IK（Ca）. Conclusion: Hyposmotic membrane stretch may activate phospholipase A2, which hydrolyzes membrane phospholipids to ultimately produce AA; AA as a second messenger mediates Ca2+influx, which triggers Ca2+-induced Ca2+release and elicits activation of IK（Ca）in gastric antral circular myocytes of the guinea pig.

关键词： gastric myocyte arachidonic acid hyposmotic membrane stretch calcium-activated potassium current.

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Relationship between atrial natriuretic peptide-immunoreactive cells and microvessels in rat gastric mucosa

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Acta Pharmacologica Sinica 2006年第2期27卷 205-211页

作者： Chun-hui LI~(2,3) Zong-wei YANG~3 Zheng-ri YIN~3 Zheng JIN~3 De-gang XING~3 Lian-hua PIAO~3 Yong-chul KIM~4 Wen-xie XU~(2,5) ~2college of Life Science and Biotechnology.Shanghai Jiaotong university,Shanghai 200240,China:~3department of physiology.yanbian university college of medicine.yanji 13.000.China ~4department of physiology,Chungbuk National university,college of medicine.Cheongju.Chungbuk 361-763,Korea collegeofLifeScienceandBiotechnology ShanghaiJiaotongUniversityShanghai200240China departmentofphysiology YanbianUniversityCollegeofMedicineYanji133000China departmentofphysiology ChungbukNationalUniversity

Aim:To investigate the ultrastructural localization of atrial natriuretic peptide(ANP)-synthesizing cells and the relationship between ANP-synthesizing cellsand microvessels in rat gastric ***:Immunohistochemistry tech-niques and postembedding immunoelectron microscopy techniques were used tovalidate the findings regarding the expression of ANP-synthesizing cells and theultrastructural localization of ANP-synthesizing cells in the gastric ***-tochemistry techniques and the tannic acid-ferric chloride method(TA-Fe stain-ing method)were used to reveal microvessel density and the distribution of ANP-synthesizing cells in different regions of the ***:Cells expressingANP were localized and ANP-synthesizing cells were identified as enterochroma-ffin(EC)cells in the gastric ***-synthesizing cells existed in differentregions of the *** percentage ANP-synthesizing cells in the mucosawas greatest in the fundus(46.7%e5.3%),intermediate in the antrum(40.1%±4.5%),and least in the body(21.6%±3.6%).There was a positive relationshipbetween the percentage of ANP-synthesizing cells and the density of microvesselsin the antral mucosa,but not in the fundus or body ***:ANP issynthesized by EC cells in rat gastric mucosa,and ANP-synthesizing cells aremost dense in the gastric *** may act not only as a regional autocrineand/or paracrine regulator,but also as an endocrine regulatory peptide in thegastrointestinal tract.

关键词： atrial natriuretic factor enterochromaffin cells blood vessels gastric mucosa

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Autoantibody against Cardiac β1-Adrenoceptor Induces Apoptosis in Cultured Neonatal Rat Cardiomyocytes

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Acta Biochimica et Biophysica Sinica 2006年第7期38卷 443-449页

作者： Yan GAO~1 Hui-Rong LIU~1 Rong-Rui ZHAO~1 Jian-Ming ZHI~(2*) 1 department of Cardiovascular physiology Shanxi Medical university,Taiyuan 030001,China 2 department of physiology,Shanghai Jiaotong university,college of Basic medicine,Shanghai 200025,China department of Cardiovascular physiology Shanxi Medical UniversityTaiyuan 030001 China department of physiology Shanghai Jiaotong University College of Basic MedicineShanghai 200025 China

To clarify whether apoptosis is involved in the injury processes induced by autoantibodyagainst cardiac β1-adrenoceptor,we investigated the biological and apoptotic effects of antibodies on culturedneonatal rat *** rats were immunized with peptides corresponding to the second extra-cellular loop of the β1-adrenoceptor to induce the production of anti-β1-adrenoceptor antibodies in the ***（Ig）G in the sera was detected using synthetic antigen enzyme-linked immunosorbentassay and purified using the diethylaminoethyl cellulose ion exchange *** of cardiomyo-cytes was evaluated using agarose gel electrophoresis and flow *** results showed that thepositive serum IgG greatly increased the beating rates of cardiomyocytes and showed an"agonist-like"***,positive serum IgG induced cardiomyocyte apoptosis after treatment with β1-adrenoceptor overstimulation for *** effects of monoclonal antibody against β-adrenoceptor werealso found to be similar to those of positive serum *** was suggested that the autoantibody could inducecardiomyocyte apoptosis by excessive stimulation of β1-adrenoceptor.

关键词： β1-adrenoceptor rat cardiomyocyte autoantibody apoptosis

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Diverse Effects of L-arginine on Cardiac Function of Rats Subjected to Myocardial Ischemia and Reperfusion in vivo

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Acta Biochimica et Biophysica Sinica 2007年第3期39卷 201-207页

作者： Xiaoliang WANG~1 Feng LIANG~2 Xiangying JIAOI Lei LIU~1 Xiaojie BAI~1 Meixia LI~1 Jianmin ZHI~3 Huirong LIU~1* 1 department of physiology,Shanxi Medical university,Taiyuan 030001,China 2 department of Cardiothoracic Surgery,General Hospital of Taiyuan Iron and Steel Company Taiyuan 030003,China 3.department of physiology.Shanghai Jiaotong university.college of Basic medicine,Shanghai 200025,China department of physiology Shanxi Medical UniversityTaiyuan 030001 China department of Cardiothoracic Surgery General Hospital of Taiyuan Iron and Steel CompanyTaiyuan 030003 China department of physiology Shanghai Jiaotong University College of Basic MedicineShanghai 200025 China

In vivo administration of L-arginine at different time points during the course of myocardialischemia and reperfusion（MI/R）has been shown to differentially regulate postischemic *** is one of the most important indexes used to judge the degree of myocardial *** presentstudy attempted to determine whether in vivo administration of L-arginine at different stages of MI/R has adiverse influence on cardiac function of ischemic reperfused hearts and,if so,to investigate the *** adult rats were subjected to 30 min myocardial ischemia followed by 5 h *** L-arginine bolus was given either 10 min before and 50 min after reperfusion（early treatment）or3 h and 4 h after reperfusion（late treatment）.Early treatment with L-arginine markedly increased the leftventricular systolic pressure（LVSP）and dP/dtmax,and decreased myocardial nitrotyrosine *** strictcontrast,late treatment with L-arginine resulted in a significant decrease in LVSP and dP/dtmaxfrom 4 h to 5h after reperfusion,and increase in toxic peroxynitrite formation as measured by *** resultssuggest that the administration of L-arginine at different time points during the course of MI/R leads todiverse effects on cardiac *** supplementation decreased the nitrative stress and improvedleft ventricular ***,late treatment with L-arginine increased the formation of peroxynitriteand aggravated cardiac functional injury.

关键词： myocardial ischemia reperfusion cardiac function nitric oxide peroxynitrite

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Synthesis and anticonvulsant activity of 1-substituted benzyl- N-substituted-1, 2, 3-triazole-4-formamides

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Progress in Natural Science:Materials International 2006年第9期16卷 925-929页

作者： WANG Junmin, JUN Changsoo, CHAI Kyuyun, KWAK Kyungchell, QUAN Zheshan ( Key Laboratory of Natural Resources of the Changbai Mountain and Functional Molecules, yanbian university, yanji 133002, China Henan university of Traditional Chinese medicine Pharmacy college, Zhengzhou 450003, China department of Chemistry, Wonkwang university, Iksan 570-749, Korea) Key Laboratory of Natural Resources of the Changbai Mountain and Functional Molecules Yanbian University Yanji 133002 China

Substituted benzyl azids were synthesized through the reaction of substituted benzyl chloride and sodium azid, which subsequently underwent cyclization with ethyl propiolate and amidation to give thirteen 1-substituted benzyl-N-substituted-1, 2, 3-triazole-4-formamide derivatives (3a-3m) . The structure of the synthesized compounds was confirmed by IR, 1H-NMR, MS and elemental analysis. Their ant icon vulsant activity against maximal electrolshock (MES) induced seizure was tested and the result showed that all these compounds possess anticonvulsant activity in different degrees. Among those, the compounds containing chloro atoms on the phenyl ring were less potent in anticonvulsant activity, while introducing one or two fluorin atoms on benzyl system increased its activity. Furthermore, their activity decreased when there was substituent on the nitrogen atom of carboxamide, and the larger the substituent, the lower the activity.

关键词： 1-substituted benzyl-N-substituted-1 2 3-triazole-4-formamide anticonvulsant synthesis.

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Antisense Oligodeoxynucleotide Inhibits Expression of Re-com binantPorcine Follicle-Stim ulating Horm one Receptor

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Journal of Huazhong university of Science and Technology(Medical Sciences) 1999年第3期19卷 175-180页

作者： ZHU Changhong 1, Mark D. Nixon 2, WANG Yifang 1, Andrew R. LaBarbera 3 1 Center of Reproductive medicine, Tongji Hospital, Tongji Medical university, Wuhan 430030 2 department of Obstetrics and Gynecology, university of Cincinnati college of medicine, Cincinnati, Ohio 45267 0526 3.department of Molecular and Cellular physiology, university of Cincinnati college of medicine, Cincinnati, Ohio 45267 0526 Center of Reproductive medicine Tongji Hospital Tongji Medical University Wuhan department of Obstetrics and Gynecology University of Cincinnati College of Medicine Cincinnati department of Molecular and Cellular physiology University of Cincinnati College of Medicine Cincinnati

To assess the role of follicle stimulating hormone receptor(FSHR) gene expression in regulating expression of FSHR protein in the plasma membrane, the effects of a porcine FSHR cDNA antisense oligodeoxynucleotide (ODN) on FSHR mRNA levels and 125 I FSH binding were determined in Chinese hamster ovary cells expression recombinant porcine FSHR (pFSHR CHO cells). An 18 mer phosphorothioate endcapped antisense ODN that corresponded to the region surrounding the translation initiation codon of the porcine FSHR cDNA was synthesized. An 18 mer nonsense sequence of identical nucleotide composition, which had little homology to known DNA sequences, was synthesized for use as a control. pFSHR CHO cells were cultured in 24 well plates (10 5 cells/well) in the absence or presence of 1 20 μmol/L antisense or nonsense ODN for 24 h and then assayed for porcine FSHR mRNA, using quantitative reverse transcription and competitive polymerase chain reaction, and for 125 I FSH binding activity. Treatment with 10 μmol/L antisense ODN caused a paradoxical increase in porcine FSHR mRNA from 0.89±0.06 to 1.64±0.08 ng/mg total RNA ( P 33 μmol/L antisense ODN caused an increase in porcine mRNA from 0.95±0.08 to 1.53±0.07 ng/mg total RAN. This was probably due to upregulation of mRNA synthesis resulting from inhibition of porcine FSHR protein translation. The nonsense ODN had no effect on porcine FSHR mRNA. Antisense, but not nonsense, ODN (10 μmol/L) inhibited membrane binding of 125 I FSH by 13.6± 0.8 % ( P 33 μmol/L antisense ODN at 0 h caused a 76.1±1.3 % ( P 3±2.3 % ( P < 0.05) of normal by 48 h. These results indicate

关键词： follicle stimulating hormone antisense oligodeoxynucleotide folliculogenesis mRNA spermatogenesis

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Astragalus polysaccharide reduces hepatic endoplasmic reticulum stress and restores glucose homeostasis in a diabetic KKAy mouse model

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Acta Pharmacologica Sinica 2007年第12期28卷 1947-1956页

作者： Xian-qing MAO~2,Yong WU~3,Ke WU~2,Ming LIU~2,Jing-fang ZHANG~4,Feng ZOU~2,Jing-ping OU-YANG~(2,5)~2 department of Pathophysiology,Medical college of Wuhan university,Hubei Provincial Key Laboratory of Allergy and Immune-RelatedDiseases,Wuhan 430071,China ~3 Division of Endocrinology and Diabetes,department of medicine,university of Oklahoma Health ScienceCenter,Oklahoma city,Oklahoma 73104,USA ~4 department of Pathology and physiology,Jingmen Vocational college,Jingmen 448000,China departmentofPathophysiology MedicalCollegeofWuhanUniversityHubeiProvincialKeyLaboratoryofAllergyandImmune-RelatedDiseasesWuhan430071China universityofOklahomaHealthScienceCenter OklahomacityOklahoma73104USA

Aim:To examine the potential effects of Astragalus polysaccharide （APS） onhepatic endoplasmic reticulum （ER） stress in vivo and in vitro and its link withhypoglycemia activity,thus establishing the mechanism underlying the hypogly-cemic action of ***:The obese and type 2 diabetic KKAy mousemodel,which is the yellow offspring of the KK mice expressed Aygene (700mg·kg-1·d-1,8 weeks) and a high glucose-induced HepG2 cell model （200 μg/mL,24 h）were treated with *** oral glucose tolerance test was measured to reflexinsulin sensitivity with the calculated homeostasis model assessment （HOMA-IR） *** 1 （XhoI site-binding protein 1） transcription and splicing,an indica-tor of ER stress,was analyzed by RT-PCR and real-time *** expression andactivation of glycogen synthase kinase 3 beta （GSK3β）,an insulin signaling protein,was measured by Western ***:APS can alleviate ER stress in cul-tured cells in *** hyperglycemia status,systemic insulin sensitivity,fattyliver disease,and insulin action in the liver of diabetic mice were partly normalizedor improved in response to APS ***:Our results indicatethat APS enables insulin-sensitizing and hypoglycemic activity at least in part byenhancing the adaptive capacity of the ER,which can further promote insulinsignal ***,APS has promising application in the treatment of type2 diabetes.

关键词： Astragalus membranaceus polysaccharide type 2 diabetes mellitus insulin resistance endoplasmic reticulum stress

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An increase in opening of BKCa channels in smooth muscle cells in streptozotocin-induced diabetic mice

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Acta Pharmacologica Sinica 2004年第6期25卷 42-48页

作者： Chun-ling YE2, Bing SHEN4, Xian-da REN, Rong-jing LUO3, Sheng-yuan DING3, Fu-man YAN3, Jia-hua JIANG department of Pharmacology, Pharmacy college of Ji-nan university, Guangzhou 510632 3department of physiology, Guangzhou university of Traditional Chinese medicine, Guangzhou 510407 4department of Molecular Biology and Cell Biology, School ofLife Sciences, university of Sciences and Technology of China, Hefei 230036, China department of Pharmacology Pharmacy College of Ji-nan University Guangzhou 510632 department of Molecular Biology and Cell Biology School of Life Sciences University of Sciences and Technology of China Hefei department of physiology Guangzhou University of Traditional Chinese Medicine Guangzhou 510407

AIM: To investigate the changes of function of large conductance of calcium-activated potassium channels (BKCa channels) in thoracic aortic smooth muscle cells in early stage of streptozotocin (STZ)-induced diabetic C57BL/6J mice. METHODS: Vascular muscle tension in the isolated thoracic aortic rings of mice was compared, and the role of BKCa channels in relaxation of isolated mice thoracic aortic rings induced by acetylcholine (ACh) was determined. Meanwhile, single vascular smooth muscle cells (VSMCs) were isolated by collagenase, and BKCa currents were recorded by patch-clamp single channel recording technique in symmetric high potassium solution. RESULTS: Tetraethylammonium (TEA) 1 mmol/L, a selective calcium-activated potassium channel blocker, caused significant rightward shift in the concentration-response curves of ACh in the isolated thoracic aortic rings of diabetic mice and pD2 value of ACh-induced relaxation was decreased notably after TEA treatment [(6.3±0.4) vs (6.9±0.5), n=10 rings from 7 mice, P0.05]. Furthermore, conductance of BKCa channels in single thoracic aortic smooth muscle cells was decreased [(199±15) pS, n=10 cells from 7 mice vs (266±11) pS, n=12 cells from 6 mice, P32±98), n=6 cells from 6 mice, P<0.05]. CONCLUSION: The opening of BKCa channels was increased in thoracic aortic smooth muscle cells in the early stage of STZ-induced diabetic C57BL/6J mice by reducing mean closed time, but the conductance of BKCa channels was decreased.

关键词： diabetes mellitus streptozotocin aorta patch-clamp techniques potassium channels calcium

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Hypoglycemic effect of Astragalus polysaccharide and its effect on PTP1B

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Acta Pharmacologica Sinica 2005年第3期26卷 345-352页

作者： Yong WU~(2,3) Jing-ping OU-YANG~(2,5) Ke WU~2 Ya WANG~2 Yun-feng ZHOU~2 Chong-yuan WEN~42 department of Pathophysiology Medical college of Wuhan university,3.department of physiology Hubei college of Traditional Chinese medicine4 department of Endocrinopathic Sciences,Renmin Hospital,Wuhan university,Wuhan 430060,China 3department of physiology Hubei College of Traditional Chinese Medicine Wuhan 430060 China 2department of Pathophysiology Medical College of Wuhan University Wuhan 430060 China 2department of Pathophysiology Medical College of Wuhan University Wuhan 430060 China 2department of Pathophysiology Medical College of Wuhan University Wuhan 430060 China 2department of Pathophysiology Medical College of Wuhan University Wuhan 430060 China 4department of Endocrinopathic Sciences Renmin Hospital Wuhan University Wuhan 430060 China

Aim:To examine the effects ofAstragalus polysaccharide (APS),a component ofan aqueous extract of Astragalus membranaceus roots,on protein tyrosine phos-phatase 1B (PTP1B),a negative regulator of insulin-receptor (IR) signaltransduction,and its potential role in the amelioration of insulin ***:Ten-week-old fat-fed streptozotocin (STZ)-treated rats,an animal modelof type Ⅱ diabetes mellitus (TIIDM),were treated with APS (400 mg/kg po) for *** sensitivity was identified by the insulin-tolerance *** analy-ses on the possible changes in insulin signaling occurring in skeletal muscle andliver were performed by immunoprecipitation or Western ***1B activ-ity was measured by an assay ***:The diabetic rats responded to APSwith a significant decrease in body weight,plasma glucose,and improved *** activity and expression of PTP1B were elevated in the skeletalmuscle and liver of TIIDM *** the insulin signaling in target tissues *** reduced both PTP1B protein level and activity in the muscle,but not in the iiver of TIIDM ***-induced tyrosine phosphorylation of theIR β-subunit and insulin receptor substrate-1 (IRS-1) were increased in the muscle,but not in the liver of APS-treated TIIDM *** was no change in the activityor expression of PTP1B in APS-treated normal rats,and blood insulin levels didnot change in TIIDM rats after treatment with ***:APS enablesinsulin-sensitizing and hypoglycemic activity at least in part by decreasing theelevated expression and activity of PTP1B in the skeletal muscles of TIIDM rats.

关键词： Astragalus membranaceus polysaccharides PTP1B type Ⅱdiabetes mellitus insulin resistance streptozotocin

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Cardioprotective mechanism of atrial natriuretic peptide

Cardioprotective mechanism of atrial natriuretic peptide

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2012年中美临床和转化医学国际论坛心脑血管病分会论坛（延吉）暨第十二届全国心血管药理学术会议和中俄药理国际研讨会

作者： Lan Hong1,Zhengai Chen2,Xun Cui1 1 department of physiology,college of medicine,yanbian university,yanji,China 2 department of Pharmacology,college of medicine,yanbian university,yanji,China.

Heart is no longer considered only as an efficient pump but also as an important endocrine organ, becausre the heart generates and secretes atrial natriuretic peptide(ANP).It is well known that ANP have important biological effects,such as diuresis and natriuresis,modulating blood pressure and cardiovascular homeostasis by combining with natriuretic peptide *** binds to NPR-A receptor which is a member of the membrane-bound guanylyl cyclase family and exerts its biological effects by increasing cGMP levels.

关键词： Atrial natriuretic peptide Mitochondrial permeability transition pore Glycogen synthase kinase 3β Myocardial ischemia/reperfusion injury Protein kinase G Phosphoinositide 3-kinase/Akt

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