内质网应激介导蜕皮甾酮对H2 O2 诱导的心肌细胞损伤的保护作用

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(H2 O2 )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;H2 O2 组,不同浓度H2 O2(1*10-3 、1*10-4 、1*10-5 mol/ L)诱导损伤细胞;EDS 组,在H2 O2 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,H2 O2 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)。H2 O2 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H2 O2 诱导的心肌细胞损伤。

Endoplasmic reticulum stress influences protective effect of ecdysterone in H2O2 induced myocardial injury

Objective:To study the role of endoplasmic reticulum stress(ERS) on ecdysterone(EDS) influenced protective effect in H2 O2 induced myocardial injury.Methods: The H9c2 cells were randomly divided into control group(normal cells),H2 O2 groups(the cells treated with H2 O2 at concentrations of 1*10-3 ,1*10-4 ,1*10-5 mol/ L,respectively);EDS group(the cells were exposed to H2 O2 and 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 levelsofBcl-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 H2 O2 group as compared with control group, which were all significantly reversed by EDS treatment ( P < 0.05).Conclusion: Ecdysterone exhibits a protective effect on thecardiomyocytes with H2 O2 induced injury by inhibiting endoplasmic reticulum stress.