中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2014, Vol. 9 ›› Issue (12): 1241-1248.doi: 10.4103/1673-5374.135333

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

microRNA-124过表达促进骨髓间充质干细胞向神经元样细胞的分化

  

  • 收稿日期:2014-05-23 出版日期:2014-06-24 发布日期:2014-06-24

Overexpression of microRNA-124 promotes the neuronal differentiation of bone marrow-derived mesenchymal stem cells

Defeng Zou 1, Yi Chen 2, Yaxin Han 1, Chen Lv 1, Guanjun Tu 1   

  1. 1 Department of Orthopedics, First Affiliated Hospital of China Medical University, Shenyang, Liaoning Province, China
    2 Department of Orthopedics, Jinhua Central Hospital of Zhejiang University, Jinhua, Zhejiang Province, China
  • Received:2014-05-23 Online:2014-06-24 Published:2014-06-24
  • Contact: Guanjun Tu, M.D., Department of Orthopedics, First Affiliated Hospital of China Medical University, Shenyang 110001, Liaoning Province, China, tu188@sina.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81070971.

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摘要：

microRNA在干细胞自我更新及其分化中发挥着重要的调节作用。所以我们设计对骨髓间充质干细胞与神经干细胞或神经元差异最明显的microRNA进行干预，增强其表达，探索促进骨髓间充质干细胞定向神经元分化的可能性。我们通过基因芯片技术，比较骨髓间充质干细胞、神经干细胞、神经元的microRNA的表达差异，挑选差异最明显的（减弱）microRNA-124进行研究。构建microRNA-124过表达慢病毒载体，以其转染骨髓间充质干细胞后，细胞中神经元早期标志物β-III 微管蛋白和微管相关蛋白2表达明显上调，细胞抗凋亡能力增强。将转染microRNA-124的骨髓间充质干细胞移植至大鼠脊髓损伤部位后，脊髓神经元标志物神经丝蛋白200在脊髓损伤部位出现大量免疫阳性表达；BBB评分显示，脊髓损伤大鼠后肢运动功能明显改善。由此，我们认为microRNA-124是骨髓间充质干细胞向神经元定向诱导分化的重要靶点，为提高骨髓间充质干细胞移植修复脊髓损伤的效力提供了新思路。

关键词: 神经再生, microRNA-124, 慢病毒, 过表达, 骨髓间充质干细胞, 神经干细胞, 脊髓损伤, 神经发生, 基因芯片, 运动功能

Abstract:

stem cells. In this study, we examined the effects of miRNA-124 (miR-124) overexpression in bone marrow-derived mesenchymal stem cells. In particular, we focused on the effect of overexpression on the differentiation of bone marrow-derived mesenchymal stem cells into neurons. First, we used GeneChip technology to analyze the expression of miRNAs in bone marrow-derived mesenchymal stem cells, neural stem cells and neurons. miR-124 expression was substantially reduced in bone marrow-derived mesenchymal stem cells compared with the other cell types. We constructed a lentiviral vector overexpressing miR-124 and transfected it into bone marrow-derived mesenchymal stem cells. Intracellular expression levels of the neuronal early markers β-III tubulin and microtubule-associated protein-2 were significantly increased, and apoptosis induced by oxygen and glucose deprivation was reduced in transfected cells. After miR-124-transfected bone marrow-derived mesenchymal stem cells were transplanted into the injured rat spinal cord, a large number of cells positive for the neuronal marker neurofilament-200 were observed in the transplanted region. The Basso-Beattie-Bresnahan locomotion scores showed that the motor function of the hind limb of rats with spinal cord injury was substantially improved. These results suggest that miR-124 plays an important role in the differentiation of bone marrow-derived mesenchymal stem cells into neurons. Our findings should facilitate the development of novel strategies for enhancing the therapeutic efficacy of bone marrow-derived mesenchymal stem cell transplantation for spinal cord injury.

Key words: nerve regeneration, microRNA-124, lentivirus, overexpression, bone marrow-derived mesenchymal stem cells, neural stem cells, spinal cord injury, neurogenesis, GeneChip, motor function, NSFC grant, neural regeneration