miR-148a促进人诱导性多能干细胞向心肌样细胞的分化  

miRNA-148a promotes the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells

作　　者：毛庆[1] 庞毅恒 卢永祥[1] 梁秀琳[1] Mao Qing;Pang Yiheng;Lu Yongxiang;Liang Xiulin(Department of Cardiology,The Second Affiliated Hospital of Guangxi Medical University,Nanning 530007,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学第二附属医院心内科

出　　处：《中国组织工程研究》2020年第19期2978-2984,共7页Chinese Journal of Tissue Engineering Research

基　　金：广西省自然科学基金项目(2017GXNSFAA198214)，项目负责人：毛庆~~

摘　　要：背景:研究发现,miR-148a可以促进人骨髓间充质干细胞定向分化为成熟心肌样细胞,但关于miR-148a对人诱导性多能干细胞向心肌样细胞分化的影响则未见报道。目的:探讨miR-148a对人诱导性多能干细胞向心肌样细胞分化的影响。方法:将人诱导性多能干细胞分为3组向心肌样细胞诱导分化,对照组细胞不做处理,低表达组细胞用miR-148a抑制物处理28 d,高表达组细胞用miR-148a模拟物处理28 d;另取常规培养28 d的人诱导性多能干细胞为空白对照组。CCK-8检测细胞增殖活力,qRT-PCR检测miR-148a mRNA的表达,免疫荧光染色和Western blot检测MHC和cTnT蛋白的表达。结果与结论:①低表达组细胞内miR-148a mRNA表达、细胞增殖活力低于空白对照组和对照组,高表达组高于其他3组(P<0.01);②空白对照组无MHC和cTnT的阳性表达,对照组、低表达组和高表达组均有MHC和cTnT的阳性表达;③低表达组细胞MHC和cTnT蛋白表达低于对照组,高表达组高于其他3组(P<0.01);④以上结果提示,miR-148a可促进人诱导性多能干细胞向心肌样细胞分化。BACKGROUND:Studies have shown that miRNA-148a can promote human bone marrow mesenchymal stem cells to differentiate into mature cardiomyocyte-like cells,but the effect of miRNA-148a on the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells has not been reported.OBJECTIVE:To investigate the effect of miRNA-148a on the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells.METHODS:Human induced pluripotent stem cells differentiating into cardiomyocyte-like cells were divided into three groups.Cells in the control group were not treated.Cells in the low expression group were treated with miRNA-148a for 28 days,and those in the high expression group were treated with mimics of miRNA-148a for 28 days.In addition,human induced pluripotent stem cells cultured for 28 days were taken as the blank control group.CCK-8 was used to detect cell proliferation activity.qRT-PCR was used to detect the expression of miRNA-148a.Immunofluorescence staining and western blot analysis were performed to detect the expression of MHC and cTnT protein.RESULTS AND CONCLUSION:The expression of intracellular miR-148a mRNA and cell proliferation activity in the low expression group were lower than those in the blank control and control groups,while those in the high expression group were significantly higher than those in the other three groups(P<0.01).There were no positive expression of MHC and cTnT in the blank control group.There were positive expressions of MHC and cTnT in the control,low expression and high expression groups.The expression of MHC and cTnT protein in the low expression group was significantly lower than that in the control group,and that in the high expression group was significantly higher than that in the other three groups(P<0.01).These results suggest that miRNA-148a can promote the differentiation of human induced pluripotent stem cells into cardiomyocyte-like cells.

关 键 词：干细胞 人诱导性多能干细胞 心肌样细胞 定向分化 miR-148a 微小RNA 人肌球蛋白重链 心脏肌钙蛋白T 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]