微小RNA-92a调控音猥因子途径对心肌缺血再灌注损伤后促血管再生的影响及机制  

Effect and mechanism of miRNA-92a regulating SHH pathway on promoting vascular regeneration after myocardial I/R injury

作　　者：朱宝华 孙艳君 李敏 Zhu Baohua;Sun Yanjun;Li Min(Department of General Medicine,Jinan People's Hos pital,Jinan 271199,Shandong Province,China)

机构地区：[1]济南市人民医院全科医学科,271199

出　　处：《中华老年心脑血管病杂志》2023年第10期1083-1087,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

摘　　要：目的探究微小RNA-92a(miR-92a)调控音猥因子(SHH)途径对心肌缺血再灌注(I/R)损伤后促血管再生的影响及机制。方法从1~3d龄SD大鼠中培养原代心肌细胞后建立心肌细胞缺氧/复氧模型,将其分成心肌细胞常氧培养组和心肌细胞I/R组。miR-92a模拟物(mimic)和拮抗剂(inhibitor)转染入大鼠原代心肌细胞,使miR-92a过表达或低表达,分成对照组、I/R组、miR-92amimic组和miR-92ainhibitor组。细胞计数法8测定各组细胞活力,流式细胞仪检测各组细胞凋亡率,ELISA和QT-PCR测定血管内皮生长因子、碱性成纤维细胞生长因子和血管生成素1等血管新生因子表达,Westernblot法测定SHH信号通路蛋白表达。结果心肌细胞I/R组miR-92a表达显著高于心肌细胞常氧培养组(3.89±0.29vs1.53±0.19,P<0.01)。对照组、miR-92ainhibitor组、I/R组和miR-92amimic组SHH蛋白(0.57±0.13vs0.51±0.11vs0.24±0.03vs0.14±0.02,P<0.01)、Smoothened蛋白(0.53±0.12vs0.49±0.10vs0.14±0.04vs0.09±0.01,P<0.01)、神经胶质瘤相关癌基因同源蛋白1(0.56±0.14vs0.50±0.13vs0.15±0.03vs0.08±0.01,P<0.01)和神经胶质瘤相关癌基因同源蛋白2(0.58±0.11 vs0.49±0.12vs0.18±0.02vs0.11±0.03,P<0.01)比较差异有统计学意义。结论miR-92a在心肌I/R损伤中异常高表达,抑制miR-92a表达可激活SHH信号通路有效促进血管再生因子表达。Objective To explore the effect and mechanism of miR-92a regulating sonic hedgehog(SHH)pathway on promoting vascular regeneration after myocardial ischemia-reperfusion(I/R)injury.Methods Primary cardiomyocytes were isolated from newborn SD rats(1 to 3 days old),and then cultured to establish a cellular model of hypoxia/reoxygenation injury.The cardiomyocytes were divided into cardiomyocyte normoxia group and cardiomyocyte I/R group.After miR-92a mimic and inhibitor were respectively transfected into primary cardiomyocytes to overexpress or lower its expression,the cells were then grouped into control,I/R,miR-92a mimic and inhibitor groups.CCK-8 assay was used to determine cell viability,flow cytometry was employed to detect cell apoptosis,ELISA and QT-PCR were applied to detect the expression of VEGF,b-FGF and Ang-1,and Western blotting was performed to measure the expression of SHH signaling pathway related proteins.Results The expression level of miR-92a was significantly higher in the cardiomyocytes from the ischemia/reperfusion(I/R)group than the normoxia group(3.89±0.29 vs 1.53±0.19,P<0.01).Statistical differences were observed among the control group,miR-92a inhibitor group,I/R group,and miR-92a mimic group in the protein levels of SHH(0.57±0.13 vs 0.51±0.11 vs 0.24±0.03 vs 0.14±0.02,P<0.01),of Smoothened(SMO,0.53±0.12 vs 0.49±0.10 vs 0.14±0.04 vs 0.09±0.01,P<0.01),of glioma-associated oncogene homolog 1(Gli-1,0.56±0.14 vs 0.50±0.13 vs 0.15±0.03 vs 0.08±0.01,P<0.01),and of glioma-associated oncogene homolog 2(Gli-2,0.58±0.11 vs 0.49±0.12 vs 0.18±0.02 vs 0.11±0.03,P<0.01).Conclusion MiR-92a is abnormally highly expressed in cardiomyocytes after I/R injury,and inhibition of miR-92a can activate SHH signaling pathway to promote the expression of angiogenesis factors effectively.

关 键 词：猬蛋白质类 心肌缺血再灌注损伤 循环微RNA 细胞低氧 细胞凋亡 微小RNA-92a 

分 类 号：R54[医药卫生—心血管疾病]