中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2015, Vol. 10 ›› Issue (2): 314-320.doi: 10.4103/1673-5374.143439

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

MicroRNA-9激活自噬活性促进大鼠骨髓间充质干细胞向神经元样细胞样细胞分化

  

  • 收稿日期:2014-08-25 出版日期:2015-02-17 发布日期:2015-02-17
  • 基金资助:

    郑州省科技厅项目（121pptgg507-11）

MicroRNA-9 promotes the neuronal differentiation of rat bone marrow mesenchymal stem cells by activating autophagy

Guang-yu Zhang 1, Jun Wang 1, Yan-jie Jia 2, Rui Han 3, Ping Li 1, Deng-na Zhu 1   

  1. 1 Rehabilitation and Treatment Center for Children with Cerebral Palsy of Henan Province, the Third Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China
    2 Department of Neurology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China
    3 Department of Pediatrics, the Third Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China
  • Received:2014-08-25 Online:2015-02-17 Published:2015-02-17
  • Contact: Deng-na Zhu, M.D., zhudengna@126.com.
  • Supported by:

    This study was supported by a grant from the Science and Technology Bureau of Zhengzhou City in China, No. 121PPTGG507-11 and the National Natural Science Foundation of China, No. 81071114, 81371385.

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摘要：

有报道称microRNA-9 (miR-9)能促进骨髓间充质干细胞向神经元样细胞分化，但具体机制尚不清楚。我们前期研究发现自噬活性增加后能够促进骨髓间充质干细胞向神经元样细胞分化，并且越来越多的证据表明miRNAs能够调节自噬通路。为此，实验设计通过miR-9-1慢病毒载体和miR-9-1抑制子调节miR-9的表达，检测LC3阳性荧光点数量及LC3-Ⅱ/LC3-Ⅰ比值评价自噬活性，检测神经元样细胞标志物神经元特异性烯醇化酶及微管相关蛋白2表达评价大鼠骨髓间充质干细胞神经分化效率。结果发现，当miR-9过表达后，LC3阳性荧光点数量及LC3-Ⅱ/LC3-Ⅰ比值增加，神经元特异性烯醇化酶及微管相关蛋白2表达增加，自噬活性提高的骨髓间充质干细胞更易于分化为神经元样细胞。说明miR-9可通过增加自噬活性促进骨髓间充质干细胞向神经元样细胞样细胞分化。

关键词: 神经再生, microRNA-9, 骨髓间充质干细胞, 分化, 神经元样细胞, 细胞自噬, 神经元特异性烯醇化酶, 微管相关蛋白, LC3

Abstract:

MicroRNA-9 (miR-9) has been shown to promote the differentiation of bone marrow mesenchymal stem cells into neuronal cells, but the precise mechanism is unclear. Our previous study confirmed that increased autophagic activity improved the efficiency of neuronal differentiation in bone marrow mesenchymal stem cells. Accumulating evidence reveals that miRNAs adjust the autophagic pathways. This study used miR-9-1 lentiviral vector and miR-9-1 inhibitor to modulate the expression level of miR-9. Autophagic activity and neuronal differentiation were measured by the number of light chain-3 (LC3)-positive dots, the ratio of LC3-II/LC3, and the expression levels of the neuronal markers enolase and microtubule-associated protein 2. Results showed that LC3-positive dots, the ratio of LC3-II/LC3, and expression of neuron specific enolase and microtubule-associated protein 2 increased in the miR-9+ group. The above results suggest that autophagic activity increased and bone marrow mesenchymal stem cells were prone to differentiate into neuronal cells when miR-9 was overexpressed, demonstrating that miR-9 can promote neuronal differentiation by increasing autophagic activity.

Key words: nerve regeneration, microRNA-9, bone marrow mesenchymal stem cells, differentiation, neuron-like cells, autophagy, neuron specific enolase, microtubule-associated protein, LC3, neural regeneration