卡维地洛对体外柯萨奇病毒B3致心肌细胞凋亡与Bcl-2/Bax蛋白表达的影响  

Influence of Carvedilol on Apoptosis and Expression of Bcl-2/Bax Protein of Cultured Cardiomyocytes Infected by Coxsackievirus B3

作　　者：席俊芳[1] 韩波[1] 陈永芬[1] 高聆[2] 马春燕[2] 张捷[2] 路康[1] 孙士静[1] 

机构地区：[1]山东大学附属省立医院小儿心脏科,济南250021 [2]山东大学附属省立医院中心实验室,济南250021

出　　处：《实用儿科临床杂志》2010年第6期412-415,共4页Journal of Applied Clinical Pediatrics

基　　金：山东省科技攻关计划项目(2009GG10002058)

摘　　要：目的探讨卡维地洛对体外柯萨奇病毒B3(CVB3)致心肌细胞凋亡的抑制作用及其机制。方法采用差速贴壁法培养原代Wistar乳鼠心肌细胞。实验分为正常对照组、病毒组、卡维地洛组及美托洛尔组。正常对照组仅加入不含血清的DMEM培养液,后3组于CVB3感染心肌细胞后分别加入不含血清的DMEM培养液、无血清培养基配制的卡维地洛、美托洛尔溶液。72h后在倒置相差显微镜下观察各组心肌细胞的形态、搏动速率;采用黄嘌呤氧化酶法测定其心肌细胞培养液中超氧化物歧化酶(SOD)活性,硫代巴比妥酸法测定其心肌细胞培养液中丙二醛(MDA)水平,流式细胞术检测其心肌细胞凋亡率,免疫细胞化学法检测其心肌细胞内Bcl-2及Bax蛋白表达水平。结果1.与病毒组比较,卡维地洛组心肌细胞搏动速率明显提高(P<0.05)、凋亡率明显降低(P<0.05),心肌细胞培养液中SOD活性显著升高(P<0.05)、MDA水平明显降低(P<0.05);美托洛尔组与病毒组比较,心肌细胞搏动速率、凋亡率、心肌细胞培养液中SOD活性、MDA水平均无统计学差异(Pa>0.05);2.与病毒组及美托洛尔组比较,卡维地洛组心肌细胞内Bcl-2蛋白表达增多,而Bax蛋白表达减弱。结论卡维地洛对体外CVB3感染的心肌细胞具有保护作用,作用机制可能与其具有抗氧化活性、增强心肌细胞内Bcl-2蛋白表达、减弱Bax蛋白表达有关。Objective To investigate the effects of carvedilol on apoptosis and expression of Bcl-2/Bax protein of cultured cardiomyocytes infected by coxsackievirus B3（CVB3）.Methods Cardiomyocytes derived from neonatal Wistar rats were purified by differential adhesion method.There were 4 groups：CVB3 group,carvedilol group,metoprolol group and normal control group.The cardiomyocytes of the first 3 groups were infected by CVB3 and then added by serum-free dulbecco＇s modified eagle medium（DMEM）,carvedilol media,metoprolol media,respectively.The cardiomyocytes in normal control group were added by serum-free DMEM.The histopathologic changes and rhythmical beat of cardiomyocytes were observed by inverted phase contrast after 72 h.Superoxide dismutase（SOD）activity and malondialdehyde（MDA）content in the culture media were measured by xanthine oxidase method and thiobarbituric acid method,respectively.Flow cytometry was used to detect cardiomyocytes apoptosis.The expression of Bcl-2 and Bax protein in cardiomyocytes were determined by immunocytochemistry.Results 1.Compared with CVB3 group,the beating rate of cardiomyocytes and SOD activity in culture media were significantly increased（Pa〈0.05）,MDA content in culture media and cardiomyocytes apoptosis were significantly decreased in carvedilol group（Pa〈0.05）.There was no significant difference in the beating rate,cardiomyocytes apoptosis,SOD activity,and MDA content between the metoprolol group and CVB3 group（Pa〉0.05）.2.The expression of Bcl-2 protein of cardiomyocyte in carvedilol group was stronger than that of CVB3 group and metoprolol group,while the expression of Bax protein of cardiomyocyte was significantly decreased.Conclusions Carvedilol is effective in inhibiting apoptosis of CVB3-infected cultured cardiomyocytes,which may be due to its antioxidation capacity,upregulation of Bcl-2 and downregulation of Bax of cardiomyocytes.

关 键 词：卡维地洛 柯萨奇病毒B3 心肌细胞 凋亡 BCL-2 BAX 

分 类 号：R96[医药卫生—药理学]