【摘要】 目的:探讨硫化氢(H2S)后处理对H9C2细胞缺氧/复氧损伤的保护作用及其机制。方法:细胞随机分为:正常对照组(con)、缺氧/复氧组(H/R)、NaHS后处理组(H/R+NaHS)、CSE阻断剂干预组(H/R+PAG)。倒置显微镜、流式细胞仪检测细胞凋亡;实时荧光定量PCR检测胱硫醚-!-裂解酶(CSE)mRNA表达;Western blotting检测p50ATF6、GRP78蛋白表达。结果:H/R组CSE mRNA表达较对照组明显降低(P<0.01);H/R+NaHS组CSE mRNA表达较H/R组明显升高(P<0.01)。H/R组p50ATF6、GRP78表达较对照组明显升高(P<0.01);NaHS后处理后p50ATF6和GRP78表达较H/R组升高(P<0.05);给予PAG后,p50ATF6、GRP78表达较H/R组降低(P<0.01)。结论:H2S后处理对H9C2细胞缺氧/复氧损伤具有保护作用,其机制可能与H2S可活化ATF6,上调GRP78表达,降低内质网应激有关。更多还原

【Abstract】 Objective:To investigate whether H2 S postconditioning protects H9C2 cells against the hypoxia/reoxygenation injury and its mechanism.Methods:H9C2cells were randomly divided into four groups:control group(con),hypoxia/reoxygenation group(H/R),H/R+200μmol/L NaHS group(H/R+NaHS)and H/R+10μmol/L PAG group(H/R+PAG).The cells morphology was observed by inverted microscope.The cell apoptosis rates were detected by PI stained flow cytometry and the expressions of CSE mRNA were determined by real-time quantitative PCR(Q-PCR).The protein expressions for p50ATF6 and GRP78were quantified by Western blotting.Results:Compared with H/R group,NaHS postconditioning could reduce the hypoxia/reoxygenation injury in H9C2 cells by observing the cells morphology and detecting the cellapoptosis rate.Compared with the control group,the expression of CSE mRNA decreased significantly in H/R group(P<0.01);but in H/R+NaHS group,CSE mRNA expression level increased significantly as compared with H/R group(P<0.01).The expression of p50ATF6 and GRP78protein were significantly higher in H/R group(P<0.01)than that in control group,but lower than that in H/R+NaHS group(P<0.05);Compared with H/R group,the expression of p50ATF6 and GRP78in H/R+PAG group reduced significantly(P<0.01).Conclusion:H2S postconditioning has a protective effect on H9C2 cells in hypoxia/reoxygenation injury.The effect may involve H2 S postconditioning,which can activate ATF6 and up-regulate the GRP78 protein expression to supress the endoplasmic reticulum stress to play aprotection.更多还原