BRD7对过氧化氢处理的心肌细胞线粒体膜电位和胞浆中Cyt C蛋白水平影响研究  

Influence of BRD7 on mitochondrial membrane potential and cytoplasmic Cyt C protein expression in cardiomyocytes treated with hydrogen peroxide

作　　者：李春晓 李艳 张静静 张勇涛 杨杰书 Li Chunxiao;Li Yan;Zhang Jingjing;Zhang Yongtao;Yang Jieshu(Department of Cardiovascular Medicine,Qingdao Central Hospital,Qingdao 266000,China)

机构地区：[1]青岛中心医院心血管内科,青岛266000 [2]青岛中心医院心职业病科,青岛266000 [3]青岛中心医院心查体中心,青岛266000

出　　处：《中国循证心血管医学杂志》2018年第10期1212-1215,共4页Chinese Journal of Evidence-Based Cardiovascular Medicine

基　　金：青岛市医药科研基金资助(2015-WJZD041)

摘　　要：目的研究溴区包含蛋白7(BRD7)对过氧化氢处理的心肌细胞线粒体膜电位和胞浆中细胞色素C(Cyt C)蛋白表达影响。方法以real-time PCR和Western blot方法检测过氧化氢处理后的心肌细胞中BRD7转录和表达水平。用BRD7 shRNA慢病毒和shRNA control慢病毒感染心肌细胞,以real-time PCR和Western blot方法检测过氧化氢条件下干扰效果。MTT测定细胞增殖活性,流式细胞术测定细胞凋亡情况,JC-1法检测线粒体膜电位,Western blot方法检测胞浆中Cyt C蛋白水平。结果过氧化氢处理后的心肌细胞中BRD7转录和表达水平升高。BRD7 shRNA慢病毒能够下调过氧化氢条件下心肌细胞中BRD7的表达水平。过氧化氢降低心肌细胞增殖活性,诱导心肌细胞凋亡,降低线粒体膜电位,增加胞浆中Cyt C蛋白水平。下调心肌细胞中BRD7表达可以拮抗过氧化氢对心肌细胞增殖活性、凋亡、线粒体膜电位及胞浆中Cyt C蛋白水平影响。结论下调BRD7提高过氧化氢条件下心肌细胞线粒体膜电位,降低胞浆中Cyt C蛋白水平,抑制细胞凋亡。Objective To study the influence of bromodomain-containing protein 7(BRD7)on mitochondrial membrane potential and expression of cytoplasmic cytochrome C(Cyt C)in cardiomyocytes treated with hydrogen peroxide.Methods The transcription and expression level of BRD7 in cardiomyocytes after treated with hydrogen peroxide were detected by using real-time fluorescence quantitative polymerase chain reaction(RT-PCR)and Western blotting assay.The cardiomyocytes were infected with BRD7 shRNA lentivirus and shRNA control lentivirus,and interventional effects in hydrogen peroxide conditions were detected by using RT-PCR and Western blotting assay.The cardiomyocyte proliferation activity was detected by using methyl thiazolyl tetrazolium test(MTT),cardiomyocyte apoptosis was determined by using flow cytometry,mitochondrial membrane potential was detected by using JC-1 method,and level of cytoplasmic Cyt C protein was detected by using Western blotting assay.Results The transcription and expression level of BRD7 in cardiomyocytes were increased after treated with hydrogen peroxide.The expression level of BRD7 shRNA lentivirus in cardiomyocytes was down-regulated in hydrogen peroxide conditions.The cardiomyocyte proliferation activity was decreased,cardiomyocyte apoptosis was induced,mitochondrial membrane potential was reduced and level of cytoplasmic Cyt C protein was increased by hydrogen peroxide.The cardiomyocyte proliferation activity,cardiomyocyte apoptosis,mitochondrial membrane potential and level of cytoplasmic Cyt C protein were antagonized by down-regulated by BRD7.Conclusion Down-regulated BRD7 can raise mitochondrial membrane potential in cardiomyocytes in hydrogen peroxide conditions,reduce level of cytoplasmic Cyt C protein,and inhibit cardiomyocyte apoptosis.

关 键 词：线粒体膜电位 溴区包含蛋白7 心肌细胞 过氧化氢 

分 类 号：R542.2[医药卫生—心血管疾病]