哈巴苷对H2O2诱导的大鼠心肌细胞H9c2氧化应激损伤的保护作用  

Protective Effect of Harpagide on H2O2-induced Oxidative Stress Injury of Rat Cardiomyocytes H9c2

作　　者：徐博[1] 郭伟[1] 刘丹平[1] 薛辉[1] XU Bo;GUO Wei;LIU Danping;XUE Hui(Shaanxi Provincial People′s Hospital,Xi′an 710068,Shaanxi,China)

机构地区：[1]陕西省人民医院

出　　处：《中西医结合心脑血管病杂志》2019年第15期2276-2281,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：陕西省卫生厅资助项目(No.2014D38)

摘　　要：目的探究哈巴苷(HG)对大鼠心肌细胞氧化应激损伤的作用。方法将H9c2细胞随机分为对照组(H9c2组)、哈巴苷组、H2O2组和H2O2+哈巴苷组,用H2O2处理细胞,复制氧化损伤模型。用哈巴苷(4μmoL)处理细胞,CCK8检测细胞增殖,Hoechst染色检测细胞凋亡,二氯荧光乙酰乙酸盐(DCF)法检测细胞内活性氧(ROS),根据试剂盒说明书检测上清液中超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽(GSH)浓度,Westernblot检测细胞凋亡相关蛋白Bcl-2、Bax、Caspase-3和Caspase-9的表达。结果与H9c2组比较,H2O2组心肌细胞增殖倍数明显降低,哈巴苷作用细胞4d后,H2O2+哈巴苷组心肌细胞增殖倍数明显高于H2O2组;同时,与H9c2组比较,H2O2组细胞凋亡率明显升高;与H2O2组比较,H2O2+哈巴苷组细胞凋亡率显著降低;H2O2还能显著诱导H9c2细胞Bax、Caspase-3和Caspase-9的表达,抑制Bcl-2表达;HG能显著减弱H2O2诱导Caspase-3和Caspase-9表达和抑制Bcl-2表达的作用;此外,H2O2组细胞内ROS活性及上清液中SOD和GSH浓度明显低于H9c2组,MDA浓度明显高于H9c2组,H2O2+哈巴苷组细胞内ROS活性及SOD和GSH浓度明显高于H2O2组,MDA浓度明显低于H2O2组。结论哈巴苷能通过抑制氧化应激抑制心肌细胞H9c2凋亡。Objective To investigate the protective effect of Harpagide(HG)on oxidative stress injury of rat cardiomyocytes H9c2 induced by H 2O 2.Methods H9c2 cardiomyocytes were divided into the control(H9c2)group,HG group,H 2O 2 group and H 2O 2+HG group.H 2O 2 was employed to establish the model of oxidative stress injury.Cardiomyocytes were treated with HG(4μmoL)and cardiomyocytes apoptosis were measured by Hoechst staining.Reactive oxygen species,the concentration of superoxide dismutase(SOD),malondiadehyde(MDA)and glutathione(GSH)in supernatant were detected by 2',7'-dichloroflurescein(DCF).The expressions of Bcl-2,Bax,Caspase-3 and Caspase-9 were detected by Western blot.Results Compared with the H9c2 group,cardiomyocytes growth was decreased in the H 2O 2 group,but it was increased in H 2O 2+HG group compared with that ina the H 2O 2 group.Compared with H9c2 group,cardiomyocytes apoptosis rate of H 2O 2 group was decreased,cardiomyocytes apoptosis rate of H 2O 2+HG group was increased compared with H 2O 2 group.H 2O 2 could significantly induce the expression of Bax,Caspase-3 and itcoued Caspase-9,and inhibit the expression of Bcl-2.The activity of ROS and concentrations of SOD and GSH were decreased than those in the H9c2 group,while MDA was significantly increased than those in the H9c2 group.The activity of ROS and concentrations of SOD and GSH in the H 2O 2+HG group were increased than those in the H 2O 2 group,while MDA was significantly decreased than those in the H 2O 2 group.Conclusion HG could inhibit apoptosis of cardiacmyocytes H9c2 by inhibiting oxidative stress.

关 键 词：细胞凋亡 心肌损伤 哈巴苷 氧化应激 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] R285.5[医药卫生—基础医学]