MicroRNA-185对血管紧张素Ⅱ介导的心肌细胞肥大和凋亡的影响  

Effect of microRNA-185 on angiotensin Ⅱ mediated cardiomyocyte hypertrophy and apoptosis

作　　者：杨莉 贺静 孙晓慧 乌宇亮[2] YANG Li;HE Jing;SUN Xiao-hui;WU Yu-liang(Department of Cardiovascular Medicine,Jinghe Industrial Park Changqing Oilfield Worker′s Hospital,Xi′an 710201,Shaanxi Province,China;Department of Cardiovascular Medicine,the First Affiliated Hospital of Xi′an Jiaotong University,Xi′an 710061,Shaanxi Province,China)

机构地区：[1]泾河工业园长庆油田职工医院心血管内科,陕西西安710201 [2]西安交通大学第一附属医院心血管内科,陕西西安710061

出　　处：《新乡医学院学报》2019年第11期1024-1029,共6页Journal of Xinxiang Medical University

摘　　要：目的探讨MicroRNA-185(miR-185)对血管紧张素Ⅱ(AngⅡ)介导的心肌细胞肥大及凋亡的影响。方法将心肌细胞系H9c2细胞随机分为对照组、AngⅡ处理组、阴性对照组和miR-185过表达组。对照组细胞采用常规培养;AngⅡ处理组细胞常规培养,并给予AngⅡ处理;阴性对照组细胞使用含miR-185阴性对照(NC)的无血清培养基处理24 h后给予AngⅡ处理;miR-185过表达组细胞使用含miR-185模拟物(miR-185 mimic)的无血清培养基培处理24 h后给予AngⅡ处理。采用实时定量聚合酶链反应检测各组细胞中miR-185、心房钠尿肽(ANP)、脑钠肽(BNP)、β-肌球蛋白重链(β-MHC)和细胞分裂周期蛋白42(CDC42)mRNA表达,应用细胞计数试剂盒-8检测各组细胞活性,细胞凋亡检测试剂盒测定各组细胞凋亡小体富计因子水平,Western blot法测定各组细胞中活化型多聚腺苷二磷酸核糖聚合梅(cleaved PARP)、活化型caspase 3和CDC42蛋白表达。结果与对照组比较,AngⅡ处理组和阴性对照组细胞中miR-185相对表达量、细胞活性显著降低(P<0.05),ANP、BNP、β-MHC和CDC42 mRNA相对表达量、细胞凋亡小体富计因子及活化型PARP、活化型caspase 3和CDC42蛋白相对表达量显著升高(P<0.05)。阴性对照组与AngⅡ处理组细胞中miR-185和ANP、BNP、β-MHC、CDC42 mRNA相对表达量、细胞活性、凋亡小体富计因子及活化型PARP、活化型caspase 3、CDC42蛋白相对表达量比较差异均无统计学意义(P>0.05)。与AngⅡ组和阴性对照组比较,miR-185过表达组细胞中miR-185相对表达量、细胞活性显著升高(P<0.05),ANP、BNP、β-MHC和CDC42 mRNA相对表达量、细胞凋亡小体富计因子及活化型PARP、活化型caspase 3、CDC42蛋白相对表达量显著降低(P<0.05)。结论miR-185可能通过下调心肌细胞中CDC42水平来减轻AngⅡ介导的心肌细胞损伤,抑制AngⅡ介导的心肌细胞肥大和凋亡,改善心肌细胞活性,从而发挥心肌保护作用。Objective To study the effect of microRNA-185(miR-185)on angiotensinⅡ(AngⅡ)-mediated cardiomyocyte hypertrophy and apoptosis。Methods Myocardial cell line H9c2 cells were randomly divided into control group,AngⅡgroup,negative control group and miR-185 overexpression group.The cells in the control group were cultured conventionally;the cells in the AngⅡgroup were treated with 0.1μmol·L-1 AngⅡfor 24 hours;the cells in the negative control group were treated with AngⅡafter transfection with serum-free medium containing miR-185 negative control for 24 hours,and cells in the miR-185 overexpression group were treated with AngⅡafter transfection with serum-free medium containing miR-185 mimic for 24 hours.The mRNA levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP)andβ-myosin heavy chain(β-MHC)and cell division cyclin 42(CDC42)were detected by real time-quantitative polymerase chain reaction(RT-qPCR).The cell viability was detected by cell counting kit-8.The levels of apoptosis cell body enrichment factors were detected by cell apoptosis detection kit.The expressions of cleaved poly ADP-ribose polymerase(PARP),cleaved caspase 3 and CDC42 protein were determined by Western blot.Results Compared with the control group,in the AngⅡgroup and negative control group,the expression of miR-185 and cell viability were significantly decreased(P<0.05),the mRNA levels of ANP,BNP,β-MHC and CDC42,apoptotic enrichment factors,and the expressions of cleaved PARP,cleaved caspase 3 and CDC42 protein were obviously increased(P<0.05).There was no significant difference in the miR-185 level,the mRNA level of ANP,BNP,β-MHC and CDC42,cell viability,apoptotic rich factor,and the expression of cleaved PARP,cleaved caspase 3 and CDC42 protein between the AngⅡgroup and negative control group(P>0.05).Compared with the AngⅡgroup and negative control group,the level of miR-185 and cell viability were significantly increased(P<0.05),and the expression of ANP,BNP,β-MHC and CDC42 mRNA,apoptotic rich facto

关 键 词：microRNA-185 血管紧张素Ⅱ 心肌肥大 细胞凋亡 细胞分裂周期蛋白42 

分 类 号：R542.2[医药卫生—心血管疾病]