miR1-2诱导骨髓间充质干细胞向心肌细胞分化过程中的Wnt/β-catenin信号通路  

miR1-2 induces the differentiation of bone marrow mesenchymal stem cells into cardiomyocytes via Wnt/beta-catenin signal pathway

作　　者：高维 李文薇[2] 任增福[2] 陈诗芸 Gao Wei;Li Wenwei;Ren Zengfu;Chen Shiyun(Haiyuan College of Kunming Medical University,Kunming 650106,Yunnan Province,China;Third Affiliated Hospital of Sun-Yat Sen University,Guangzhou 510000,Guangdong Province,China)

机构地区：[1]昆明医科大学海源学院,云南省昆明市650106 [2]中山大学附属第三医院,广东省广州市510000

出　　处：《中国组织工程研究》2019年第1期7-12,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:骨髓间充质干细胞具有向心肌细胞分化的潜能,但其分化率较低,且其分化机制尚不完全明确。目的:探讨miR1-2在小鼠骨髓间充质干细胞分化成心肌样细胞中的作用,揭示此过程中涉及的信号通路。方法:用miR1-2和5-氮杂胞嘧啶处理小鼠骨髓间充质干细胞。通过q PCR检测心肌细胞标志物GATA4在骨髓间充质干细胞中的表达。流式细胞术检测细胞凋亡率,并通过测定该信号通路的上游蛋白评价Wnt/β-catenin信号通路的活性。结果与结论:(1)miR1-2模拟物转染骨髓间充质干细胞24 h后,miR1-2在骨髓间充质干细胞中的表达显著增加。miR-2模拟物处理48 h后,骨髓间充质干细胞的凋亡率无显著差异;(2)转染miR1-2模拟物48 h后,与对照组相比,GATA4的表达随着时间的延长显著增加,处理72 h后,这些基因在miR1-2组中的表达显著高于5-氮杂胞嘧啶组和对照组;(3)与miR1-2-mimics-NC相比,用miR1-2模拟物处理后,β-catenin和Wnt11的m RNA和蛋白相对表达均显著增加,而添加Wnt/β-catenin信号通路抑制剂LGK-974后,β-catenin和Wnt11的相对表达量均显著降低;(4)结果提示,骨髓间充质干细胞中miR1-2可以更有效地诱导骨髓间充质干细胞向心肌细胞分化,并且与Wnt/β-catenin信号通路活性有关。BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)have the potential to differentiate into cardiomyocytes,but exhibit a low differentiation rate.Moreover,its differentiation mechanism is still unclear.OBJECTIVE:To investigate the role of miR1-2 in the differentiation of mouse BMSCs into cardiomyocytes and to reveal the signaling pathways involved in this process.METHODS:Mouse BMSCs were treated with miR1-2 and 5-azacytosine.The cardiomyocyte marker GATA4 in BMSCs was detected by qPCR.Apoptosis in BMSCs was detected by flow cytometry and the activity of Wnt/β-catenin signaling pathway was evaluated by measuring the upstream protein of this signal pathway.RESULTS AND CONCLUSION:After 24 hours of transfection with miR1-2 mimics,the expression of miR1-2 in BMSCs was significantly increased.After 48 hours of transfection with miR-2 mimics,there was no significant difference in the apoptotic rate of BMSCs.After 48 hours of transfection with miR1-2 mimics,the expression of GATA4 increased significantly with time.After 72 hours of transfection with miR1-2 mimics,the expression of these genes was significantly higher in the miR1-2 group than in the 5-azacytosine group and the control group.Compared with miR1-2-mimics-NC,treatment with miR1-2 mimics could significantly increase the relative expression ofβ-catenin and Wnt11 at mRNA and protein levels,while addition of Wnt/β-catenin pathway inhibitor LGK-974 reduced the relative expression ofβ-catenin and Wnt11.To conclude,miR1-2 in BMSCs has the potential to effectively induce the differentiation of BMSCs into cardiomyocytes via the Wnt/β-catenin pathway

关 键 词：miR1-2 骨髓间充质干细胞 心肌细胞分化 WNT/Β-CATENIN信号通路 5-氮杂胞嘧啶 凋亡率 GATA-4 细胞转染 干细胞 间充质干细胞 肌细胞 心脏 细胞分化 组织工程 

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