比索洛尔预处理对缺氧/复氧诱导的H9c2细胞损伤的影响  

Effects of bisoprolol pretreatment on hypoxia/reoxygenation-induced injury in H9c2 cardiomyocytes

作　　者：习明明[1] 刘晶[2] 谢亮[2] 宫剑滨[2] XI Mingming LIU Jing XIE Liang GONG Jianbin(Department of Cardiology, Taizhou People＇s Hospital, Taizhou 225300, China Department of Cardiology, Jinling Hospital, Nanjing University School of Medicine/Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, China)

机构地区：[1]泰州市人民医院内科,江苏泰州225300 [2]南京大学医学院附属金陵医院(南京军区南京总医院)心脏内科,江苏南京210002

出　　处：《南方医科大学学报》2016年第9期1198-1203,共6页Journal of Southern Medical University

基　　金：国家自然科学基金(81400238)~~

摘　　要：目的研究比索洛尔预处理对缺氧/复氧诱导的大鼠心肌细胞损伤的保护作用,并探讨该作用的机制。方法对大鼠心肌细胞株(H9c2)进行缺氧复氧(6 h/2 h),模拟大鼠心肌缺血再灌注损伤模型,将H9c2细胞随机分成4组:正常对照组(control组)、缺氧/复氧组(H/R组)、缺氧/复氧+比索洛尔预处理组(H/R+B组)、缺氧/复氧+比索洛尔预处理+PI3K/AKT阻滞剂LY294002组(H/R+B+LY组);分别采用MTT法检测心肌细胞活力,流式细胞仪法检测心肌细胞凋亡、ROS水平,Western免疫印迹法检测心肌细胞p-AKT及p-GSK3β蛋白表达情况。结果与正常组相比,H/R组细胞活力下降,细胞凋亡率增加、ROS水平升高;经比索洛尔预处理后,细胞活力明显提高,细胞凋亡率下降、活性氧水平降低,p-AKT/GAPDH、p-GSK3β/GAPDGH显著提高,而加入PI3K/AKT阻滞剂LY294002后比索洛尔的保护作用消失。结论比索洛尔能减少缺氧/复氧所引起的氧化应激,对心肌细胞具有明显的保护作用,其作用机制是通过激活PI3K/AKT/GSK3β通路而提高AKT、GSK3β磷酸化水平,减少ROS产生,增加细胞的活性实现的。Objective To investigated the effects of bisoprolol pretreatment on hypoxia/reoxygenation （H/R）-induced injury in H9c2 cardiomyocytes. Methods Cultured H9c2 cells were exposed to hypoxia for 6 h followed by reoxygenation for 2 h with or without pretreatments with bisoprolol or bisoprolol＋LY294002. The cell survival was measured by MTT assay, and the cell apoptosis and levels of radical oxygen species （ROS） were evaluated with flow cytometry. The protein levels of phosphyorylated AKT and phosphorylated GSK3βin the cells were determined by Western blotting. Results Compared with the normal control cells, the cells exposed to H/R injury showed significantly decreased cell survival and increased cell apoptosis and ROS production; pretreatment of the cells with bisoprolol significantly decreased the cell apoptotic rates and ROS production and obviously enhanced the cell survival and protein levels of p-AKT and p-GSK3βin the exposed cells. The protective effect of bsioprolol against H/R-induced cell injury was significantly attenuated by LY294002. Conclusion Bisoprolol can protect H9c2 cells against H/R-induced injury and oxidative stress by activating PI3K/AKT/Gsk-3βpathway to increase the phosphorylation of AKT and GSK3βand reduce ROS production.

关 键 词：比索洛尔 缺氧/复氧损伤 H9C2 心肌保护 PI3K/AKT GSK3Β 

分 类 号：R542.2[医药卫生—心血管疾病]