吡那地尔预处理对缺氧/复氧成后年大鼠心肌细胞内钙离子浓度的影响  

Effect of Pinacidil preconditioning on calcium concentration of cultured adult rat cardiomyocytes during hypoxia/reoxygenation

作　　者：邓胜利[1] 姚刚[1] 张琳[1] 喻田[1] 

机构地区：[1]贵州省遵义医学院麻醉学系,563003

出　　处：《贵州医药》2010年第12期1059-1061,共3页Guizhou Medical Journal

基　　金：贵州省教育厅自然科学研究培育项目(编号:黔教科(2007)038号);遵义医学院硕士启动基金(编号:院字(2007)20号)

摘　　要：目的观察吡那地尔(Pinacidil)预处理对缺氧/复氧后成年大鼠心肌细胞内Ca2+浓度([Ca2+]i)的影响,以探讨吡那地尔在缺氧/复氧中抑制钙超载作用的机制。方法通过Langen-dorff离体心脏灌注装置,用Ⅱ型胶原酶对心脏逆行灌注,进行成年大鼠心肌细胞的分离与培养,建立心肌细胞缺氧/复氧模型。实验分组及处理:(1)正常组(NOR组):细胞正常培养;(2)缺氧/复氧组(CON组):细胞在95%N2+5%CO2培养箱中缺氧45 min,复氧1 h;(3)吡那地尔组(Pina组):先加入吡那地尔(30μmol/L)预处理15 min,再缺氧45 min,复氧1h;(4)格列苯脲拮抗Pina组(Gi+Pina组):先用格列苯脲(50μmol/L)处理5 min后,余处理同Pina组。通过Fluo-3/AM荧光指示剂负载,用激光共聚焦显微镜检测心肌细胞Ca2+浓度变化。结果 NOR组心肌细胞内钙离子荧光强度值较低;细胞缺氧45 min,复氧1 h后其荧光强度值与NOR组相比显著增高(P<0.01);而Pina组细胞内荧光强度值较CON组及Gi+Pina组低(P<0.01)。结论心肌细胞缺氧/复氧后可导致细胞内钙超载,而吡那地尔能明显减轻心肌细胞缺氧/复氧后的钙超载现象,其机制可能与ATP敏感性钾通道的开放有关。Objective To investigate the effect of Pinacidil preconditioning on calcium concentration of cultured adult rat cardiomyocytes after hypoxia/reoxygenation,and to search for the mechanism of repressed intracellular calcium concentration by Pinacidil.Methods Adult rat cardiomyocytes were separated and cultured by Langendorff isolated heart perfusion apparatus.The cultured cells were randomly divided into 4 groups which were normal group(NOR),control group(CON),Pinacidil preconditioning group(Pina) and Glibenclamide plus Pinacidil group(Gi+Pina).The untreated cultured cells were continuous cultured in group NOR.In group CON,oxygen was deprived for 45 min and was restored for 60 min.In group Pina,30μmol/L pinacidil preconditioning for 15 min was executed before hypoxia,and the rest procedure was carried out in the same way as group CON.In group Gi+Pina,the cultured cells were treated with 50μmol/L Glibenclamide for 5 min before pinacidil preconditioning,and the rest was same as group Pina.The calcium concentrations of cultured cardiomyocytes of adult rat were observed by the Confocal microscope.ResultsThe inflorescences intensity of cardiomyocytes were significantly lower in NOR group than that in CON group and Gi+Pina group(P<0.01).The inflorescences intensity of cardiomyocytes were significantly lower in the Pina group than that in Gi+Pina group and CON group(P<0.01).Conclusion Pinacidil preconditioning can decrease intracellular calcium concentration that was up regulated markedly in hypoxia/ reoxygenation,which may be related to the opening of ATP sensitive potassium channel.

关 键 词：吡那地尔 缺氧/复氧 心肌细胞 钙 

分 类 号：R96[医药卫生—药理学]