高糖诱导心肌细胞miR-26a/miR-197表达促进细胞凋亡的机制研究  

Expressions of miR-26a/miR-197 induced by high glucose promoting cell apoptosis in cardiomyocytes:a study on mechanism

作　　者：刘祖霞 张斌 汪伟 张康林 黄军利 朱小山[2] 杨峰[2] Liu Zuxia;Zhang Bin;Wang Wei;Zhang Kanglin;Huang Junli;Zhu Xiaoshan;YangFeng(Department of Cardiology,People's Hospital (Third Affiliated Hospital of Jianghan University),HuangpiDistrict,Wuhan 430300,China.)

机构地区：[1]武汉市黄陂区人民医院江汉大学附属第三医院心内科,武汉430300 [2]襄阳市中心医院心内科湖北文理学院附属医院,襄阳441021

出　　处：《中国循证心血管医学杂志》2019年第5期551-555,共5页Chinese Journal of Evidence-Based Cardiovascular Medicine

基　　金：湖北省自然科学基金青年项目(2016CFB344)

摘　　要：目的探讨miR-26a/miR-197在高糖诱导的心肌细胞表达及对细胞活力和凋亡率的影响。方法 H9c2心肌细胞分为低糖组(LG组)、LG+miRcramble组、高糖(HG)+miRcramble组、HG+miR-26ainhibitor组、HG+miR-197 inhibitor组和HG+miR-26a/miR-197 inhibitor组,RT-PCR检测细胞中miR-26a和miR-197的表达;MTT法及流式细胞术分别检测细胞活力及凋亡率;Western blotting检测β-catenin、PCNA和Bax的蛋白表达。通过靶基因预测软件、miR-26a/miR-197的mimics/inhibitor转染后的细胞中MCL1表达检测及miR-NC、Wt-MCL1+miR-26a mimics、Wt-MCL1+miR-197 mimics、Mut-MCL1+miR-26a mimics和Mut-MCL1+miR-197 mimics共转染后的荧光素酶活性检测,确定MCL1与miR-26a/miR-197是否存在靶向关系。结果 HG诱导的H9c2心肌细胞miR-26a和miR-197的表达均明显高于LG组,而转染miR-26a/miR-197 inhibitor后miR-26a和miR-197表达均明显低于HG组(P<0.05)。HG+miRcramble组细胞活力及PCNA的蛋白表达均明显低于LG+miRcramble组,细胞凋亡率及β-catenin和Bax的蛋白表达均明显高于LG+miRcramble组(P<0.05),而HG+miR-26a inhibitor组和HG+miR-197 inhibitor组细胞活力及PCNA的蛋白表达均明显高于HG+miRcramble组,低于HG+miR-26a/miR-197 inhibitor组,细胞凋亡率及β-catenin和Bax的蛋白表达均明显低于HG+miRcramble组,高于HG+miR-26a/miR-197 inhibitor组(P<0.05)。MCL1是miR-26a和miR-197的靶基因。结论 miR-26a/miR-197可通过靶向MCL1促进高糖诱导心肌细胞活力和降低细胞凋亡率,机制可能与抑制Wnt信号通路有关。Objective To investigate the influence of Expressions of miR-26a/miR-197 induced by high glucose (HG) in cardiomyocytes on cell viability and apoptosis rate. Methods H9c2 cardiomyocytes were divided into low glucose group (LG group),LG+miRcramble group,HG+miRcramble group,HG+miR-26a inhibitor group,HG+miR-197 inhibitor group and HG+miR-26a/miR-197 inhibitor group. The expressions of miR- 26a and miR-197 were detected by using RT-PCR,and cell viability and apoptosis rate were detected by using methyl thiazolyl tetrazolium test (MTT) and flow cytometry (FCM). The protein expressions of β-catenin,PCNA and Bax were detected by using Western blotting assay. The target relationship between MCL1 and miR-26a/ miR-197 was determined through target gene prediction software,detection of MCL1 expression after transfection of miR-26a/miR-197 mimics/inhibitor,and detection of luciferase activity after co-transfection of miR-NC,Wt- MCL1+miR-26a mimics,Wt-MCL1+miR-197 mimics,Mut-MCL1+miR-26a mimics and Mut-MCL1+miR-197 mimics. Results The expressions of miR-26a and miR-197 induced by HG in H9c2 cardiomyocytes were significantly higher in all HG groups than those in LG group,and significantly lower in HG+miR-26a/miR-197 inhibitor group than those in other HG groups (P<0.05). The cell viability and PCNA protein expression were significantly lower,and apoptosis rate and protein expressions of β-catenin and Bax were significantly higher in HG+miRcramble group than those in LG+miRcramble group (P<0.05). The cell viability and PCNA protein expression were significantly higher in HG+miR-26a inhibitor group and HG+miR-197 inhibitor group than those in HG+miRcramble group,and lower than those in HG+miR-26a/miR-197 inhibitor group. The apoptosis rate and protein expressions of β-catenin and Bax were significantly lower in HG+miR-26a inhibitor group and HG+miR-197 inhibitor group than those in HG+miRcramble group,and higher than those in HG+miR-26a/ miR-197 inhibitor group (P<0.05). MCL1 was the target gene of miR-26a and miR-197. Concl

关 键 词：miR-26a miR-197 心肌细胞 高糖 凋亡 WNT信号通路 

分 类 号：R542.2[医药卫生—心血管疾病]