冠心康对大鼠缺血心肌细胞ATP敏感钾通道亚基Kir6.1、Kir6.2、SUR2A和SUR2B表达的影响  

Effects of Guanxinkang on expressions of ATP-sensitive potassium channel subunits Kir6.1,Kir6.2,SUR2A and SUR2B in ischemic myocytes of rats

作　　者：陈富荣[1] 张娜[1] 刘萍[1] 章怡祎[1] 韩向晖[1] 蔡珏峰[1] 

机构地区：[1]上海中医药大学龙华医院科技处,上海200032

出　　处：《中西医结合学报》2010年第5期458-464,共7页Journal of Chinese Integrative Medicine

基　　金：上海市教育委员会重点项目(No.07ZZ56);国家教育部科学技术研究重点项目(No.208042)

摘　　要：目的:通过观察中药复方冠心康对大鼠缺血心肌细胞ATP敏感钾(ATP-sensitive potassium,K_(ATP))通道亚基的影响,探讨冠心康保护心血管、抗心肌缺血的可能作用机制。方法:48只SPF级Wistar大鼠随机分为正常组、模型组、格列本脲组、吡那地尔组、冠心康组、冠心康加格列本脲组。采用酶解法分离大鼠心室肌细胞,用无钙台式液灌流10 min、停灌30 min、再灌注45 min模拟心肌缺血再灌注损伤。将药物直接加入细胞液中(格列本脲10μmol/L、吡那地尔50μmol/L、冠心康注射液1 ml/L)充分作用后,4℃放置24 h。采用实时荧光定量多聚酶链式反应法及蛋白质印迹法检测各组心肌细胞K_(ATP)亚基Kir6.1、Kir6.2、SUR2A和SUR2B的mRNA表达及蛋白含量。结果:正常大鼠心肌细胞可见SUR2A、Kir6.1和Kir6.2蛋白及mRNA的表达,而SUR2B蛋白几乎不表达。模型组K_(ATP)各个亚基蛋白及mRNA的表达均较正常组有一定程度的增加。与模型组比较,吡那地尔可显著增加K_(ATP)各亚基mRNA及蛋白的表达,格列本脲可阻断这一作用;冠心康可明显增加K_(ATP)通道各亚基mRNA及蛋白的表达。结论:复方冠心康具有明显的促进K_(ATP)开放的作用,从而起到心血管保护作用。Objective：To observe the effects of Guanxinkang injection,a compound traditional Chinese herbal medicine, on ATP-sensitive potassium（KATP） channel subunits in ischemic myocardial cells of rats,and to explore the mechanism of Guanxinkang in protecting myocardial ischemic reperfusion injuries. Methods：Forty-eight Wistar rats were randomly divided into normal group,untreated group,glibenclamide group,pinacidil group,Guanxinkang group and Guanxinkang plus glibenclamide group.The ventricular myocytes were prepared from hearts of normal rats by enzymatic dissociation method.The ischemic ventricular myocytes underwent perfusion with normal Tyrode＇ solution for 10 min,then stopping perfusion 30 min,and followed by 45 min of reperfusion.The glibenclamide,pinacidil and Guanxinkang were added into ventricular myocytes solution directly.Then the solutions were placed at 4℃.After 24-hour freezing at—80℃,mRNA and protein expressions of KATP subunits Kir6.1,Kir6.2,SUR2A and SUR2B were measured by real-time quantitative polymerase chain reaction and Western blotting respectively. Results：In normal rat myocardial cells,there were SUR2A,Kir6.1,and Kir6.2 protein and gene expressions but no expression of SUR2B protein.In the untreated group,all subunit mRNA and protein expressions of KATP increased to some extent as compared with the normal group.Pinacidil,a potassium channel opener, significantly increased mRNA and protein expressions of KATP subunits,while the blocker glibenclamide had a reverse effect.Meanwhile,Guanxinkang injection significantly increased mRNA and protein expressions of KATP subunits but with no significant difference as compared with pinacidil. Conclusion：Guanxinkang injection can obviously enhance the open of KATP channel and thus play a role in cardiovascular protection.

关 键 词：中成药 肌细胞 心脏 再灌注损伤 ATP敏感钾通道 

分 类 号：R285.5[医药卫生—中药学]