内质网应激介导蜕皮甾酮对H2O2诱导的心肌细胞损伤的保护作用

摘要

目的:探讨蜕皮甾酮(EDS)如何通过调节内质网应激对过氧化氢(H2O2)诱导的心肌细胞损伤起保护作用.方法:大鼠H9c2心肌细胞随机分为对照(Control)组,正常心肌细胞;H2O2组,不同浓度H2O2(1×10-3、1×10-4、1×10-5mol/L)诱导损伤细胞;EDS组,在H2O2组基础上,加入不同浓度的EDS(25、50、100 μmol/L)处理.MTT法和流式细胞术分别检测实验各组细胞活力及细胞凋亡率.Western blot检测各组细胞中 Bcl-2、Bax、cleaved-caspase-3、caspase-4、caspase-7、caspase-12、GRP78和CHOP的蛋白水平,同时检测各组细胞中丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性.结果:与Control组相比,H2O2组的细胞活力减弱,凋亡率升高,MOD含量升高,SOD活性降低,GRP78和CHOP的表达升高(均P<0.05).H2O2组加入EDS处理后,细胞活力提升,凋亡率下降,MOD含量降低,SOD活性升高,GRP78和CHOP的表达降低(均P<0.05).结论:通过抑制内质网的应激过程,蜕皮甾酮能减轻H2O2诱导的心肌细胞损伤.%Objective:To study the role of endoplasmic reticulum stress(ERS)on ecdysterone(EDS)influenced protective effect in H2O2induced myocardial injury.Methods: The H9c2 cells were randomly divided into control group(normal cells),H2O2 groups(the cells treated with H2O2at concentrations of 1×10-3,1×10-4,1×10-5mol/L,respectively);EDS group(the cells were exposed to H2O2and treated with EDS at concentrations of 25,50,100 μmol/L respectively).The cell activity and apoptosis were measured by MTT assay and flow cytometry,respectively.The protein levels of Bcl-2,Bax,cleaved-caspase-3,caspase-4,caspase-7, caspase-12,GRP78 and CHOP were deter mined by Western blot.The MDA content and SOD activity were detected by the MDA and SOD detection kits.Results:The cell activity was decreased,cell apoptosis was increased,the content of MDA was elevated,the activity of SOD was inhibited and the protein expression of GRP78 and CHOP were increased in H2O2group as compared with control group, which were all significantly reversed by EDS treatment(P<0.05).Conclusion: Ecdysterone exhibits a protective effect on the cardiomyocytes with H2O2induced injury by inhibiting endoplasmic reticulum stress.