中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (2): 345-349.doi: 10.4103/1673-5374.290908

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

miR-132/甲基CpG结合蛋白2调节神经干细胞分化中的作用

  

  • 出版日期:2021-02-15 发布日期:2020-12-04
  • 基金资助:
    国家自然科学基金项目(81271990)

Role of miR-132/methyl-CpG-binding protein 2 in the regulation of neural stem cell differentiation

Dong Chen, Jie Liu, Zhong Wu*, Shao-Hua Li*   

  1. Department of Orthopedics, Shanghai Tenth People’s Hospital, Tongji University, Shanghai, China
  • Online:2021-02-15 Published:2020-12-04
  • Contact: Zhong Wu, MD, zhongwu0515@163.com; Shao-Hua Li, MD, doctorlish77@163.com.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81271990 (to SHL).

摘要:

甲基CpG结合蛋白2(MeCP2)是众所周知的转录抑制因子,其突变会引起严重的神经系统疾病。大量研究表明MeCP2仅与神经成熟有关,尚无报道探索其在神经干细胞分化中的作用。此次实验对MeCP2在神经干细胞中的这种可能作用进行分析,使用2种不同的促分化方法来探索MeCP2如何影响神经干细胞分化。结果发现当MeCP2过表达慢病毒转染神经干细胞时,星形胶质细胞分化受到损害。即使在存在有利于星形细胞分化的20%胎牛血清条件下,这种损伤也会发生。此外,在几种中枢神经系统相关的miRNA中,miR-132在分化后存在最明显的表达变化。荧光素酶测量证实miR-132可直接靶向MeCP2,并且其能够降低MeCP2 mRNA和蛋白的表达。而miR-132模拟物可上调miR-132表达并促进星形胶质细胞的分化,但这种作用可被MeCP2过表达则能完全恢复。表明miR-132可通过减少MeCP2表达调节神经干细胞分化。实验已于2018年3月10日经同济大学附属上海市第十人民医院伦理委员会批准,批准号SHDSYY-2018-4748。

https://orcid.org/0000-0001-7111-621X (Shao-Hua Li)

关键词: 干细胞, 模型, 因子, 通路, 星形胶质细胞

Abstract: Methyl-CpG-binding protein 2 (MeCP2) is a well-known transcription repressor, and mutations in MECP2 cause serious neurological disorders. Many studies have suggested that MeCP2 is involved in neural maturation only, and have not reported its role in neural stem cell differentiation. In the present study, we investigated this possible role of MeCP2 in neural stem cells. We used two different differentiation methods to explore how MeCP2 influences neural stem cell differentiation. When we transfected MeCP2-overexpressing lentivirus into neural stem cells, astrocytic differentiation was impaired. This impaired astrocytic differentiation occurred even in conditions of 20% fetal bovine serum, which favored astrocytic differentiation. In addition, miR-132 had the largest expression change after differentiation among several central nervous system related miRNAs. A luciferase assay confirmed that miR-132 directly targeted MeCP2, and that miR-132 was able to reduce MeCP2 expression at both the RNA and protein levels. The upregulation of miR-132 by miRNA mimics promoted astrocytic differentiation, which was fully recovered by MeCP2 overexpression. These results indicate that miR-132 regulates cell lineage differentiation by reducing MeCP2. The study was approved by the Ethics Committee of Shanghai Tenth People’s Hospital of TongJi University, China (approval No. SHDSYY-2018-4748) on March 10, 2018.

Key words: astrocytes, factor, model, pathways, stem cell