lncRNA RP11-1C8.5通过结合miR-181a-5p对糖尿病心肌细胞凋亡和增殖的影响  

Effects of lncRNA RP11-1C8.5 on Apoptosis and Proliferation of Diabetic Cardiomyocytes by Binding to miR-181a-5p

作　　者：黄锐 周源 贺清 白杨[4] 周礼平 HUANG Rui;ZHOU Yuan;HE Qing(Department of Cardiovascular Medicine,Lichuan People′s Hospital,Hubei 471009,China)

机构地区：[1]利川市人民医院心血管内科,445400 [2]利川市人民医院肾病内分泌科,445400 [3]利川市人民医院中医科,445400 [4]华中科技大学附属同济医院心血管内科,武汉430030

出　　处：《医学研究杂志》2022年第11期93-98,共6页Journal of Medical Research

基　　金：湖北省卫生健康委员会中医药科研项目青年人才项目(ZY2021Q004)。

摘　　要：目的探究长链非编码(long non-coding RNA,lncRNA)RP11-1C8.5对糖尿病心肌细胞凋亡和增殖的影响及其分子机制。方法分别采用5.5、10.0、13.9、22.2和33.3mmol/L葡萄糖浓度的培养基培养人心肌HCM细胞24h后,实时定量聚合酶链反应(qPCR)技术检测不同葡萄糖浓度培养下HCM细胞中RP11-1C8.5的表达水平。选择RP11-1C8.5表达最高的葡萄糖浓度继续培养HCM细胞,分别感染RP11-1C8.5干扰腺病毒(实验组)和对照病毒(对照组)。流式细胞术和CCK-8法检测对照组和实验组HCM细胞的凋亡情况和细胞活力。通过生物信息学和双荧光素酶报告基因实验验证RP11-1C8.5与miR-181a-5p的结合作用。qPCR检测对照组和实验组HCM细胞中miR-181a-5p的表达水平。Western blot法检测下调RP11-1C8.5表达对凋亡相关蛋白Bax、caspase3、Bcl-2、CED-9表达的影响。结果与正常葡萄糖浓度(5.5mmol/L)比较,在高糖培养心肌HCM细胞中RP11-1C8.5的表达水平显著增加(P<0.01),其中在33.3mmol/L葡萄糖浓度培养HCM细胞中的表达最高(P<0.01)。与对照组比较,实验组HCM细胞中RP11-1C8.5的表达显著降低(P<0.01)。与对照组比较,实验组HCM细胞的凋亡率显著降低(P<0.01),细胞活力明显增加(P<0.05)。双荧光素酶报告基因实验证实RP11-1C8.5与miR-181a-5p存在互补结合作用(P<0.01)。与对照组比较,实验组HCM细胞促凋亡蛋白Bax、caspase3表达降低,抗凋亡蛋白Bcl-2、CED-9表达增加。结论下调RP11-1C8.5通过结合miR-181a-5p发挥抑制糖尿病心肌细胞凋亡和促进细胞增殖的作用。Objective To explore the effect of long non-coding RNA(lncRNA)RP11-1C8.5 on apoptosis and proliferation of diabetic cardiomyocytes and its molecular mechanism.Methods After culturing human myocardial HCM cells with glucose concentrations of 5.5,10.0,13.9,22.2 and 33.3mmol/L for 24h,real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of RP11-1C8.5 in HCM cells cultured with different glucose concentrations.The glucose concentration with the highest expression of RP11-1C8.5 was selected to continue to culture HCM cells,and were infected with RP11-1C8.5 interfering adenovirus(experimental group)and control virus(control group),respectively.Flow cytometry and CCK-8method were used to detect the apoptosis and cell viability of HCM cells in the control and experimental groups.The binding effect of RP11-1C8.5 and miR-181a-5p was verified by bioinformatics and dual luciferase reporter gene experiments.The expression levels of miR-181a-5p in HCM cells in the control and experimental groups were detected by qPCR.Western blot was used to detect the effect of down-regulation of RP11-1C8.5 expression on the expressions of apoptosis-related proteins Bax,caspase3,Bcl-2 and CED-9.Results Compared with normal glucose concentration(5.5mmol/L),the expression level of RP11-1C8.5 in myocardial HCM cells cultured with high glucose was significantly increased(P<0.01),and the expression level was highest in HCM cells cultured at 33.3mmol/L glucose concentration(P<0.01).Compared with the control group,the expression of RP11-1C8.5 in HCM cells of the experimental group was significantly decreased(P<0.01).Compared with the control group,the apoptosis rate of HCM cells in the experimental group was significantly decreased(P<0.01),and the cell viability was significantly increased(P<0.05).The dual-luciferase reporter gene assay confirmed that RP11-1C8.5had complementary binding to miR-181a-5p(P<0.01).Compared with the control group,the expression of pro-apoptotic proteins Bax and caspase3 in HCM cells in t

关 键 词：糖尿病心肌病 长链非编码RNA miR-181a-5p 凋亡 增殖 

分 类 号：R542.2[医药卫生—心血管疾病]