上调Uqcrc1表达在过氧化氢诱导的心肌细胞氧化应激中的作用  

Role of up-regulation of Uqcrc1 expression in hydrogen peroxide-induced oxidative stress in cardiomyocytes

作　　者：梁霄 柏小川[1] LIANG Xiao;BO Xiaochuan(Department of Cardiology,the First People′s Hospital of Zunyi,Zunyi,Guizhou 563000)

机构地区：[1]遵义市第一人民医院心血管内科

出　　处：《郑州大学学报（医学版）》2019年第5期724-728,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：贵州省科技合作计划项目(黔科合LH字7123号)

摘　　要：目的:探讨泛醇-细胞色素C还原酶核心蛋白1(Uqcrc1)在过氧化氢(H2O2)诱导的心肌细胞氧化应激中的作用。方法:心肌H9c2细胞分成H9c2组、H2O2组(H2O2处理)、Lv-NC+H2O2组(感染阴性对照慢病毒并经H2O2处理)、Lv-Uqcrc1+H2O2组(感染Uqcrc1过表达慢病毒并经H2O2处理12 h)共4组。应用qRT-PCR和Western blot法测定4组细胞中Uqcrc1的表达,CCK-8测定细胞增殖,二硝基苯肼显色法检测乳酸脱氢酶(LDH)漏出率,硫代巴比妥酸法检测丙二醛(MDA)含量,黄嘌呤氧化法检测超氧化物歧化酶(SOD)活性,钼酸铵比色法检测过氧化氢酶(CAT)活性,ELISA法检测谷胱甘肽过氧化物酶(GSH-Px)活性,双染法检测细胞凋亡,Western blot检测细胞中激活型Caspase-3和Caspase-9蛋白表达水平。结果:Lv-Uqcrc1+H2O2组细胞中Uqcrc1表达水平高于H2O2组、Lv-NC+H2O2组。与H9c2组细胞比较,H2O2组细胞增殖活性降低,LDH漏出率升高,细胞中CAT、SOD、GSH-Px活性降低,MDA含量升高,细胞凋亡率升高,细胞中Caspase-3和Caspase-9蛋白表达水平升高(P<0.05)。与H2O2组和Lv-NC+H2O2组比较,Lv-Uqcrc1+H2O2组细胞增殖活性升高,LDH漏出率降低,细胞中CAT、SOD、GSH-Px活性升高,MDA含量降低,细胞凋亡率降低,细胞中Caspase-3和Caspase-9蛋白表达水平降低(P<0.05)。结论:Uqcrc1能减轻H2O2诱导的心肌细胞氧化应激损伤。Aim:To study the role of ubiquinol-cytochrome C reductase core protein 1(Uqcrc1)in hydrogen peroxide(H2O2)-induced oxidative stress in cardiomyocytes.Methods:H9c2 cells were divided into H9c2 group(control group),H2O2 group(H2O2 treatment),Lv-NC+H2O2 group(infected with negative control lentivirus and treated with H2O2),and Lv-Uqcrc1+H2O2 group(infected with Uqcrc1 overexpression lentivirus vector and treated with H2O2 for 12 h).qRT-PCR and Western blot were used to measure the expression of Uqcrc1,cell proliferation was detected by CCK-8 assay,the leakage rate of LDH was detected by dinitrophenylhydrazine colorimetry,the content of MDA was detected by thiobarbituric acid,the activity of SOD was detected by xanthine oxidation,the CAT activity was detected by ammonium molybdate colorimetric method,the GSH-Px activity was detected by ELISA,apoptosis was detected by double staining,the expression levels of activated Caspase-3 and Caspase-9 protein were detected by Western blot.Results:The expression level of Uqcrc1 in Lv-Uqcrc1+H2O2 group was higher than that in H2O2 group and Lv-NC+H2O2 group.Compared with control group,the proliferation activity of H2O2 group decreased,the leakage rate of LDH increased,the activity of CAT,SOD and GSH-Px decreased,the content of MDA increased,the rate of apoptosis increased,and the levels of Caspase-3 and Caspase-9 protein in cells increased(P<0.05).Compared with H2O2 group and Lv-NC+H2O2 group,the proliferation activity of Lv-Uqcrc1+H2O2 group increased,the leakage rate of LDH decreased,the activity of CAT,SOD and GSH-Px increased,the content of MDA decreased,the apoptosis rate decreased,the levels of Caspase-3 and Caspase-9 protein in cells decreased(P<0.05).Conclusion:Uqcrc1 could alleviate oxidative stress induced by hydrogen peroxide in cardiomyocytes.

关 键 词：Uqcrc1 心肌细胞 氧化应激 凋亡 H9C2细胞 

分 类 号：R542.2[医药卫生—心血管疾病]