miR⁃93⁃5p对心脏成纤维细胞纤维化的影响  

Effects of miR⁃93⁃5p on fibrosis of cardiac fibroblasts

作　　者：张伟峰[1] 雷素扬[1] 赵俊涛[1] ZHANG Weifeng;LEI Suyang;ZHAO Juntao(Department of Cardiology,Zhengzhou Seventh People's Hospital,Zhengzhou,Henan,China,450000)

机构地区：[1]郑州市第七人民医院心外科,河南郑州450000

出　　处：《分子诊断与治疗杂志》2020年第5期656-660,共5页Journal of Molecular Diagnostics and Therapy

基　　金：河南省医学科技攻关计划项目(2018063215)。

摘　　要：目的miR⁃93⁃5p是心肌梗死中新发现的差异微小RNA(miRNA),探索其在心脏成纤维细胞中的功能机制。方法用差速贴壁法分离心肌梗死小鼠心脏成纤维细胞和心肌细胞;脂质体法将miR⁃NC组(转染miR⁃NC)、miR⁃93⁃5p mimic组(转染miR⁃93⁃5p mimic)、inhibitor⁃NC组(转染inhibitor⁃NC)、miR⁃93⁃5p inhibitor组(转染miR⁃93⁃5p inhibitor)、pcDNA组(转染pcDNA)、pcDNA⁃弗林蛋白酶(Furin)组(转染pcDNA⁃Furin)、si⁃NC组(转染si⁃NC)、si⁃Furin组(转染si⁃Furin)、miR⁃93⁃5p mimic+pcD⁃NA组载体(共转染miR⁃93⁃5p mimic和pcDNA)、miR⁃93⁃5p mimic+pcDNA⁃Furin组(共转染miR⁃93⁃5p mimic和pcDNA⁃Furin)转染至分离的成纤维细胞。实时荧光定量逆转录聚合酶链反应(qRT⁃PCR)、免疫印迹(Western blot)检测细胞中miR⁃93⁃5p、Furin、胶原蛋白Ⅰ(CollagenⅠ)、胶原蛋白Ⅲ(CollagenⅢ)、成纤维细胞基质金属蛋白酶抑制物⁃2蛋白(TIMP2)、纤维连接蛋白(Fibronectin)的表达;双荧光素酶报告基因检测实验检测细胞的荧光活性。结果心肌梗死小鼠心脏成纤维细胞与心肌细胞相比,miR⁃93⁃5p、Furin的表达均显著升高(P<0.05)。过表达miR⁃93⁃5p、敲减Furin能明显抑制成纤维细胞中CollagenⅠ、CollagenⅢ、TIMP2、Fibronectin的表达,而抑制miR⁃93⁃5p、过表达Furin则具有相反的作用。miR⁃93⁃5p直接抑制野生型Furin细胞的荧光素酶活性,并且过表达Furin还可部分逆转miR⁃93⁃5p对心脏成纤维细胞的纤维化调控。结论miR⁃93⁃5p可抑制心肌梗死心脏成纤维细胞的纤维化,其机制与靶向Furin有关。Objective To explore the molecular function of miR⁃93⁃5p in cardiac fibroblasts.Methods The cardiac fibroblasts and cardiomyocytes of myocardial infarction mice were isolated by differen⁃tial adherence method.The miR⁃NC group(transfected miR⁃NC),miR⁃93⁃5p mimic group(transfected miR⁃93⁃5p mimic),inhibitor⁃NC group(transfected inhibitor⁃NC),miR⁃93⁃5p inhibitor group(transfected miR⁃93⁃5p inhibitor),pcDNA group(transfected pcDNA),pcDNA⁃Furin group(transfected pcDNA⁃Furin),si⁃NC group(transfected si⁃NC),si⁃Furin group(transfected si⁃Furin),miR⁃93⁃5p mimic+pcDNA group vector(co⁃transfected miR⁃93⁃5p mimic and pcDNA),miR⁃93⁃5p mimic+pcDNA⁃Furin group(co⁃transfected miR⁃93⁃5p mimic and pcDNA⁃Furin)were transfected into isolated fibroblasts.qRT⁃PCR and Western blot were used to detect the expression of miR⁃93⁃5p,Furin,CollagenⅠ,CollagenⅢ,TIMP2,and Fibronectin in the cells.Dual luciferase reporter assay was used to detect the fluorescence activity of the cells.Results The ex⁃pression of miR⁃93⁃5p and Furin was significantly higher in cardiac fibroblasts than in cardiomyocytes(P<0.05).Overexpression of miR⁃93⁃5p and knockdown of Furin significantly inhibitethe expression of CollagenⅠ,CollagenⅢ,TIMP2,and Fibronectin in cardiac fibroblasts,while inhibition of miR⁃93⁃5p and overexpression of Furin had opposite effects.miR⁃93⁃5p directly inhibits the luciferase activity of wild⁃type Furin cells,and overexpression of Furin can also partially reverse the fibrosis of cardiac fibroblasts by miR⁃93⁃5p.Conclusions miR⁃93⁃5p can inhibit fibrosis of cardiac fibroblasts in myocardial infarction,and its mecha⁃nism is related to targeting Furin.

关 键 词：miR⁃93⁃5p FURIN 心肌梗死 成纤维细胞纤维化 

分 类 号：R542.22[医药卫生—心血管疾病]