卡维地洛通过PI3K/AKT/GSK3β信号通路对H9c2缺氧/复氧损伤的保护  

Carvedilol protect H9c2 against hypoxia/reoxygenation injury via PI3K/AKT/GSK3β pathway

作　　者：习明明[1] 刘晶[1] 邹莺[1] 杭涛[1] 汤沂[1] 谢亮[1] 李言明[1] 宫剑滨[1] 

机构地区：[1]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,210002

出　　处：《心肺血管病杂志》2016年第4期317-321,共5页Journal of Cardiovascular and Pulmonary Diseases

基　　金：国家青年科学基金项目基金资助(81400238)

摘　　要：目的:研究卡维地洛预处理对缺氧/复氧诱导的大鼠心肌细胞保护作用及其机制。方法:对大鼠心肌细胞株(H9c2)进行缺氧/复氧(6h/2h),模拟大鼠心肌缺血再灌注损伤模型,将H9c2细胞随机分成四组:正常对照组(control组)、缺氧/复氧组(H/R组)、缺氧/复氧+卡维地洛预处理组(H/R+CAR组)、缺氧/复氧+卡维地洛+PI3K/AKT阻滞剂LY294002组(H/R+CAR+LY294002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活性氧(ROS)水平,Western免疫印迹法检测心肌细胞Caspase-3、p-AKT、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,ROS水平升高,Caspase-3表达明显增多;经卡维地洛预处理后,细胞活力明显提高、p-AKT、p-GSK3β磷酸化水平较H/R组显著增加,ROS水平明显降低,而加入PI3K/AKT阻滞剂LY294002后卡维地洛的上述作用显著减弱。结论:卡维地洛预处理能显著减轻缺氧/复氧诱导的心肌细胞损伤,对心肌细胞具有明显的保护作用,其可能的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,增加细胞的活性而实现的。Objective： To investigated the protective mechanisms of carvedilol pretreatment on H9c2 during hypoxia / reoxygenation injury. Methods： Cultured H9c2 cell was randomly divided into 4 groups： normal control group（ control）; hypoxia / reoxygenation group（ H / R）; hypoxia / reoxygenation ＋ carvedilol group（ H/R ＋ CAR）; hypoxia/reoxygenation ＋ carvedilol ＋ LY294002 group（ H/R ＋ CAR ＋ LY294002）. The H/R procedure was peformed for 6 hours hypoxia and 2 hours reoxygenation in H9c2 cells. The viability of H9c2 cell was measured by MTT,radical oxygen species（ ROS） rates were evaluated by flow cytometry. The protein levels of Caspase-3 and phosphyorylated AKT（ p-AKT） and phosphyorylated GSK3β（ p-GSK3β） were determined by Western Blotting. Results： Compared with control group,the cell viability was significantly decreased,ROS and Caspase-3 levels were increased in H / R group; whereas compared with H / R group,ROS and the Caspase-3 levels were reduced,and the cell viability and the protein levels of p-AKT and p-GSK3β were enhanced markedly in the H / R ＋ CAR group. Using the PI3 K / AKT inhibitor LY294002,the protect effect of carvedilol was weakened obviously. Conclusions： Carvedilol could protect the cardiomyocyte from hypoxia / reoxygenation induced injury. Its mechanism is involved in the activation of PI3 K / AKT / GSK3β survival pathway via increasing the phosphorylation of AKT and GSK3β,which lead to increasing cell viability and reducing the production of ROS.

关 键 词：卡维地洛 预处理 缺氧/复氧损伤 H9C2 心肌保护 

分 类 号：R54[医药卫生—心血管疾病]