吡那地尔、美托洛尔、谷氨酰胺、胰岛素单独及联合使用对H9c2心肌细胞缺氧/复氧损伤的抗凋亡作用及机制  

Antiapoptotic effects and mechanism of pinacidil,metoprolol, glutamine,insulin single and combination on H9c2 cardiac myocytes exposed to simulated hypoxia/reoxygenation

作　　者：孙畅[1] 臧婵媛[1,2] 吴艳娜[1] 温克[1] 康毅[1] 娄建石[1] 

机构地区：[1]天津医科大学药理学教研室,天津300070 [2]天津医学高等专科学校,天津300222

出　　处：《中国药理学通报》2011年第12期1723-1727,共5页Chinese Pharmacological Bulletin

基　　金：高等学校博士学科点专项科研基金(No 20070062009)

摘　　要：目的研究吡那地尔(pinacidil,Pin)、美托洛尔(metoprolol,Met)、谷氨酰胺(glutamine,Glu)、胰岛素(insu-lin,Ins)4药联用对缺氧/复氧(hypoxia/reoxygenation,H/R)所致H9c2心肌细胞损伤的抗凋亡保护作用并探讨其作用机制。方法将培养的H9c2心肌细胞随机分为7组,即①正常对照(Con)组;②缺氧/复氧(H/R)组;③吡那地尔(Pin)组;④美托洛尔(Met)组;⑤谷氨酰胺(Glu)组;⑥胰岛素(Ins)组;⑦吡那地尔、美托洛尔、谷氨酰胺和胰岛素4药联用(PMGI)组。细胞经药物处理建立H/R损伤模型,并测定各组H9c2心肌细胞的存活率;收集培养液测定乳酸脱氢酶(lactate dehydrogenase,LDH)活性;流式细胞仪检测心肌细胞凋亡百分率;罗丹明123(Rhodamine123,Rh123)荧光探针标记线粒体,检测荧光强度以反映心肌细胞线粒体膜电位(mitochondrial membrane potential,△Ψm)变化。结果对于H/R损伤的H9c2心肌细胞,吡那地尔、美托洛尔、谷氨酰胺与胰岛素4药联用组与药物单用组或H/R组相比能明显提高细胞存活率,保护其细胞膜,减少LDH渗漏;减少细胞凋亡率;提高△Ψm。结论吡那地尔、美托洛尔、谷氨酰胺和胰岛素4药联用后,通过不同途径保护心肌细胞,达到协同抗凋亡作用。Aim To investigate protective effects and mechanism of pinacidil,metoprolol,glutamine,insulin on H9c2 cardiac myocytes exposed to simulated hypoxia/reoxygenation.Methods H9c2 cardiac myocytes were randomly divided into seven groups：① Control group;② Hypoxia/reoxygenation（H/R） group;③ Pinacidil（Pin） group;④ Metoprolol（Met） group;⑤ Glutamine（Glu） group;⑥ Insulin（Ins） group;⑦ Pinacidil＋Metoprolol＋Glutamine＋Insulin（PMGI） group.The survival rates of cardiomyocytes were detected.The activity of lactate dehydrogenase（LDH） in the culture medium was measured.The mitochondria were labeled by Rhodamine123 fluorescence probe and the fluorescent intensity produced by Rh123 was measured,which reflected changes of mitochondrial membrane potential（MMP,△Ψm）.The apoptotic percentage was measured with flow cytometry.Results The combined use of pinacidil,metoprolol,glutamine,and insulin could protect the H9c2 cardiac cell membrane caused by hypoxia/reoxygenation injury,increase cell survival rate,decrease LDH leakage,decrease the apoptotic rate and increase the mitochondrial membrane potential.Conclusions Compared to the individual drug,the combination of the different mechanisms protects the mitochondria of cardiac myocyte,decreases the injury and enhances antiapoptotic effection of myocardial cells.

关 键 词：缺氧/复氧损伤 联合用药 心肌细胞存活率 乳酸脱氢酶 细胞凋亡率 线粒体膜电位 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]