MiR-210-3p转染对大鼠骨髓基质干细胞成骨活性影响  

Effect of Mir-210-3p transfection in rat bone marrow mesenchymal stem cells on their osteogenic capacity

作　　者：杨梦 原福贞 邓荣辉 翟小青 刘泽众 余家阔[2] YANG Meng;YUAN Fu-zhen;DENG Rong-hui;ZHAI Xiao-qing;LIU Ze-zhong;YU Jia-kuo(School of Clinical Medicine,Weifang Medical University,Weifang 261053,China;Institute of Sports Medicine,Peking University Third Hospital,Beijing 100191,China)

机构地区：[1]潍坊医学院临床学院,山东潍坊261053 [2]北京大学第三医院运动医学研究所,北京100191

出　　处：《中国矫形外科杂志》2022年第21期1975-1981,共7页Orthopedic Journal of China

基　　金：国家自然科学基金项目(编号:51773004、51920105006、52003008)。

摘　　要：[目的]探讨微小RNA(microRNA,miR)-210-3p转染大鼠骨髓基质干细胞(bone marrow mesenchymal stem cells,BMMSCs)对成骨活性的影响。[方法]体外培养大鼠BMMSCs,随机分3组,分别为空白对照组、miR-210-3p增强组(增强组)、miR-210-3p抑制组(抑制组),给予相应体外转染处理。检测细胞增殖、凋亡和迁移,以及诱导成骨分化和诱导成脂分化情况;采用qRT-PCR检测相应标志物的mRNA表达水平。[结果]第3代BMMSCs的CD44和CD90呈阳性表达,CD34和CD45呈阴性表达,符合间充质干细胞的表面抗原特征。与空白对照组对比,增强组的miR-210-3p的mRNA表达显著增加(P<0.05),而抑制组的miR-210-3p的mRNA表达显著抑制(P<0.05)。细胞增殖OD值、Transwell细胞迁移、茜素红染色钙盐沉积计数由高至低依次均为增强组>空白对照组>抑制组(P<0.05);而凋亡率和油红O染色脂滴计数由低至高依次为增强组<空白对照组<抑制组(P<0.05)。q RT-PCR检测,成骨表达基因,包括ALP和Bglap-2的mRNA相对表达量由高至低依次为增强组>空白对照组>抑制组(P<0.05);而成脂表达基因,包括PPARγ和LPL的mRNA相对表达量由低至高依次为增强组<空白对照组<抑制组(P<0.05)。[结论]本研究表明miR-210-3p可促进大鼠BMMSCs成骨分化,抑制成脂分化,减少细胞凋亡并促进细胞增殖和迁移。[Objective]To explore the effect of microRNA(miR-210-3p)transfection in rat bone marrow mesenchymal stem cells(BMMSCs)on their osteogenic capacity.[Methods]Rat BMMSCs were isolated and cultured in vitro,and were randomly divided into 3groups.The cells in the blank control group,MiR-210-3p enhanced group(enhanced group)and miR-210-3p inhibited group(inhibited group)were given transfection with the corresponding genes in vitro respectively.The cell biological behavious were detected,involving proliferation,apoptosis and migration,as well as osteogenic differentiation and adipogenic differentiation,in addition,qRT-PCR was used to detect mRNA expression levels of corresponding markers.[Results]The third generation BMMSCs obtained in this study were positive for CD44 and CD90,negative for CD34 and CD45,which were consistent with the surface antigen characteristics of mesenchymal stem cells.Compared with blank control group,mRNA expression of miR-210-3p in enhanced group was significantly increased(P<0.05),whereas which in the inhibition group was significantly inhibited(P<0.05).The OD value of cell proliferation,Transwell cell migration and alizarin red staining calcium deposition count were significantly ranked from high to low as the enhanced group>blank control group>inhibition group(P<0.05).Conversely,the apoptosis rate and oil red O staining lipid drop count were significantly ranked from low to high as the enhanced groupblank control group>inhibition group(P<0.05),whereas the mRNA relative expression levels of lipid-expressing genes,including PPARγ and LPL,were from low to high as the enhanced group<blank control group<inhibition group(P<0.05).[Conclusion]In this study the miR-210-3p does promote osteogenic differentiation,inhibit adipogenic differentiation,reduce apoptosis,and promote cell proliferation and migration of BMMSCs in rats.

关 键 词：微小RNA(miRNAs) 骨髓基质干细胞 成骨分化 成脂分化 

分 类 号：R318[医药卫生—生物医学工程]