小鼠间充质干细胞脂向分化过程中miRNA-143的表达

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (1): 12-15.

小鼠间充质干细胞脂向分化过程中miRNA-143的表达

凌宏艳¹,胡小波¹,奉水东¹,杨丝丝¹,何剑琴¹,张恺芳¹,廖端芳²

1. 南华大学
2. 湖南中医药科大学

收稿日期:2011-03-21 修回日期:2011-08-22 出版日期:2012-01-05 发布日期:2011-12-28
通讯作者: 廖端芳 E-mail:dfliao@yahoo.com.cn
基金资助:
胰岛素抵抗相关miRNAs的鉴定及功能研究;Insigs与SCAP/SREBP失平衡在高糖高胰岛素诱导巨噬细胞胆固醇蓄积中的作用及机制

The expression of miRNA-143 during adipogenic differentiation of MSCs

Received:2011-03-21 Revised:2011-08-22 Online:2012-01-05 Published:2011-12-28

摘要/Abstract

摘要： 目的：探讨小鼠间充质干细胞（MSCs）脂向分化过程中miRNA-143（miR-143）的表达，为阐明MSCs脂向分化的调控机制提供新线索。方法：采用全骨髓体外分离结合差速贴壁法纯化扩增C57BL／6小鼠MSCs，将第5代MSCs采用脂肪细胞分化诱导剂进行成脂诱导。运用miRNA芯片技术比较MSCs组和脂肪细胞组中miR-143的表达，并通过实时定量PCR技术验证。结果：成功分离、纯化、培养MSCs；MSCs经脂肪诱导剂诱导后，胞内大量脂滴形成，油红O染色阳性。MiRNA芯片及实时定量PCR结果均表明miR-143在MSCs脂向分化过程中显著上调（3.73±0.42 vs 1.00±0.14，P<0.01）。结论：MiR-143可能参与调控MSCs的脂向分化。

关键词: miRNA-143, 间充质干细胞, 脂向分化, miRNA芯片, 实时定量PCR

Abstract: Objective: To investigate the expression of miR-143 during adipogenic differentiation of Mesechymal stem cells（MSCs）in mice, and the regulation mechanisms of MSCs’ adipogenic differentiation. Methods：The MSCs of C57BL／6 mice were isolated, cultured by using the whole bone marrow method, amplified by the differential adherent method, and adipogenic differentiation of the fifth generation of MSCs was induced with the adipogenic medium. We used the microRNA microarray to explore the expression of miR-143 in the MSCs group and adipocytes group, and validated it by real-time quantitative polymerase chain reaction (real-time PCR). Results：We got high-purity MSCs and Oil O staining showed that lipid droplets were increased in MSCs treated with adipogenic conditions, which implied that the MSCs were induced into adipocyte successfully. The miR-143 was up-regulated during adipogenic differentiation showed by microRNA microarray analysis and confirmed by using real -time PCR（3.73±0.42 vs 1.00±0.14，P<0.01）.Conclusion: MiRNA-143 may take part in regulating the adipogenic differentiation of MSCs.

Key words: miRNA-143, mesenchymal stem cells, adipogenic differentiation, microRNA microarray, real-time PCR

中图分类号:

凌宏艳胡小波奉水东杨丝丝何剑琴张恺芳廖端芳. 小鼠间充质干细胞脂向分化过程中miRNA-143的表达[J]. 基础医学与临床, 2012, 32(1): 12-15.

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