下调miR-711靶向促进SIRT6表达抑制LPS诱导心肌细胞凋亡的影响和机制研究  

The effects and action mechanism of down regulating miR711 on LPS induced cardiomyocyte apoptosis in vitro

作　　者：王艳[1] 颜治 詹洮 WANG Yan;YAN Zhi;ZHAN Tao(Department of Cardiovascular Medicine,The Second People’s Hospital of Yibin City,Sichuan,Yibing 644000,China)

机构地区：[1]四川省宜宾市第二人民医院心血管内科,644000

出　　处：《河北医药》2021年第10期1455-1459,共5页Hebei Medical Journal

摘　　要：目的研究下调miR 711对脂多糖(LPS)诱导心肌细胞凋亡的影响和机制。方法以心肌细胞H9c2作为实验对象,分成Control(空白对照)、LPS(LPS处理)、Anti NC+LPS(转染inhibitor control后给予LPS处理)、Anti miR 711+LPS(转染miR 711 inhibitor后给予LPS处理)组,Realtime PCR测定miR 711表达,MTT测定细胞活力,流式细胞术检测细胞凋亡,Western blot测定Bax、Bcl2蛋白表达。生物信息学软件分析预测miR 711的靶基因,荧光素酶报告系统鉴定沉默信息调节因子2相关酶6(SIRT6)与miR 711的靶向关系。将SIRT6 siRNA和miR 711 inhibitor共转染到心肌细胞中,给予LPS处理,检测细胞活力、凋亡变化,验证miR 711作用机制。结果与Control组比较,LPS组细胞活力下降,细胞凋亡率升高,细胞中Bax蛋白表达量增加,Bcl2蛋白表达量降低,差异均有统计学意义(P<0.05)。与Anti NC+LPS组比较,Anti miR 711+LPS组细胞活力升高,细胞凋亡率降低,细胞中Bax蛋白表达量减少,Bcl2蛋白表达量增加,差异均有统计学意义(P<0.05)。miR 711靶向负调控SIRT表达。与Anti miR 711+LPS+si NC组比较,Anti miR 711+LPS+si SIRT6组细胞活力降低,凋亡率升高,Bax蛋白表达水平升高,Bcl2、SIRT6蛋白表达水平降低,差异均有统计学意义(P<0.05)。SIRT6 siRNA逆转下调miR 711对LPS条件下心肌细胞活力和凋亡影响。结论下调miR 711靶向促进SIRT6表达抑制LPS诱导的心肌细胞凋亡。Objective To investigate the effects and action mechanism of down regulating miR711 on lipopolysaccharides(LPS)induced cardiomyocyte apoptosis.Methods The cardiomyocyte H9c2 were divided into control group,LPS group,anti NC+LPS group(LPS treatment after transfecting inhibitor control),anti miR 711+LPS group(LPS treatment after transfection of miR 711 inhibitor).The miR 711 expressions were measured by Rrealtime PCR,the cell viability was detected by MTT,the apoptosis was detected by flow cytometry,and the expression levels of Bax and Bcl2 protein were determined by Western Blot.Bioinformatics soft analysis was performed to predict the target gene of miR 711,the luciferase reporter system was used to identify the targeting relationship between silent information regulation 2 homolog 6(SIRT6)and miR 711.Cotransfect SIRT6 siRNA and miR 711 inhibitor were co-transfected into cardiomyocytes,and were treated by LPS,the changes of cell viability and apoptosis were detected to verify the action mechaniasm of miR 711.Results As compared with those in control group,the cell viability in LPS group was significantly decreased,the apoptosis rate was significantly increased,the expression levels of Bax protein were significantly increased,however the expression levels of Bcl2 protein were significantly decreased(P<0.05).As compared with those in anti NC+LPS group,the cell viability in anti miR 711+LPS group was significantly increased,the apoptosis rate was significantly decreased,the expression levels of Bax protein were significantly decreased,however the expression levels of Bcl2 protein were significantly increased(P<0.05).The miR 711 could negatively targetedly regulat the SIRT expressions.As compared with those in anti miR 711+LPS+si NC group,the cell viability in anti miR 711+LPS+si SIRT6 was significantly dicreased,the apoptosis rate was significantly increased,the expression levels of Bax protein were significantly increased,the expression levels of Bcl 2 and SIRT6 were significantly decreased(P<0.05).SIRT6 siRNA coul

关 键 词：miR-711 脂多糖 心肌细胞 凋亡 

分 类 号：R542.2[医药卫生—心血管疾病]