基础医学与临床 ›› 2011, Vol. 31 ›› Issue (1): 41-46.

• 研究论文 • 上一篇    下一篇

microRNA-1诱导大鼠骨髓间充质干细胞向心肌样细胞分化

文通,魏云峰,王梦洪,汪泱,曾俊义,彭小平,李宾公   

  1. 南昌大学第一附属医院
  • 收稿日期:2010-03-17 修回日期:2010-05-11 出版日期:2011-01-05 发布日期:2011-01-05
  • 通讯作者: 魏云峰

miRNA-1 induces rat bone marrow mesenchymal stem cells differentiation into cardiomyocyte-like cells

WEN Tong 1,WEI Yun-feng 1,WANG Meng-hong 2,WANG Yang 2,ZENG Jun-yi 2,PENG Xiao-ping 2,LI Bin-gong 2   

  1. 1. The First Affiliated Hospital of Nanchang University2.
  • Received:2010-03-17 Revised:2010-05-11 Online:2011-01-05 Published:2011-01-05
  • Contact: WEI Yun-feng

摘要: 目的 研究microRNA-1(miRNA-1)能否诱导骨髓间充质干细胞(MSCs)向心肌样细胞分化。方法 构建大鼠miRNA-1表达载体,分离扩增培养及鉴定大鼠MSCs。脂质体法转染大鼠第4代MSCs,实时定量RT-PCR(qRT-PCR)检测转染miRNA-1质粒后MSCs的miRNA-1表达水平。分别于转染2、4和6天后:RT-PCR检测心肌重要转录因子GATA4、NKx2.5和MEF2C的mRNA表达,免疫荧光检测心肌特异蛋白I(cTnI)的表达。结果90%以上的MSCs表达MSCs重要标志物CD29、CD44;未检测到造血前体细胞标志抗原CD34、白细胞标志抗原CD45的表达。转染miRNA-1质粒后,miRNA-1表达水平明显上调。转染miRNA-1质粒2、4和6天后,GATA4、NKx2.5和MEF2C的mRNA表达逐渐增强。第4和6天后可见cTnI阳性表达细胞。结论 miRNA-1能诱导大鼠MSCs向心肌样细胞分化。

关键词: miRNA-1, 骨髓间充质干细胞, 心肌样细胞

Abstract: Objective To investigate whether miRNA-1 may induce rat bone marrow mesenchymal stem cells (rMSCs) differentiation into cardiomyocyte-like cells. Methods rmiRNA-1 expression vector was constructed, rMSCs were isolated, amplified and identified. The fourth passage of rMSCs were transfected miRNA-1 plasmid with Lipofectamine2000 and miRNA-1 expression was then detected by quantitative real-time RT-PCR (qRT-PCR). The mRNA expression of cardiac-specific transcription factor GATA4, NKx2.5 and MEF2C were detected by RT-PCR,cTnI expression was measured by immunofluorescence,respectively,2,4,6 days. Results More than 90% of the cultured rMSCs were positive for CD29 and CD44, which are two significant markers of MSCs and negative for hematopoetic precursor cell markers CD34,leukocyte antigen CD45. After transfection of miRNA-1 in MSCs, the expression level of miRNA-1 was markedly enhanced. The mRNA expression of GATA4, NKx2.5 and MEF2C gradually increased from day 2 to day 6. The cTnI expression was detected by day 4,6. Conclusion miRNA-1 can induce MSCs differentiation into cardiomyocyte-like cells.

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