中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (17): 2711-2716.doi: 10.3969/j.issn.2095-4344.0517

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

miR-146b促进诱导多能干细胞向神经元样细胞的分化

李 强1，张明伟1，李建明2，胡俊林2   

  1. 1西南医科大学附属中医医院神经外科，四川省泸州市 646000；2自贡市第三人民医院神经外科，四川省自贡市 643020
  • 修回日期:2018-01-30 出版日期:2018-06-18 发布日期:2018-06-18
  • 通讯作者: 张明伟，硕士生导师，西南医科大学附属中医医院神经外科，四川省泸州市 646000
  • 作者简介:李强，男，1986年生，四川省自贡市人，2012年成都医学院毕业，医师，主要从事中西医结合神经外科相关研究。

miR-146b facilitates the differentiation of induced pluripotent stem cells into neuron-like cells

Li Qiang1, Zhang Ming-wei1, Li Jian-ming2, Hu Jun-lin2   

  1. 1Department of Neurosurgery, Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China; 2Department of Neurosurgery, Zigong Third People’s Hospital, Zigong 643020, Sichuan Province, China
  • Revised:2018-01-30 Online:2018-06-18 Published:2018-06-18
  • Contact: Zhang Ming-wei, Master’s supervisor, Department of Neurosurgery, Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • About author:Li Qiang, Physician, Department of Neurosurgery, Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, Luzhou 646000, Sichuan Province, China

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摘要：

文章快速阅读：

文题释义：
miR-146b：miRNAs是一类长20-25个核苷酸的非编码RNA，成熟的miRNA通过与特定mRNA的3’端非编码区结合，从而对其剪切或者抑制其蛋白水平的表达。miRNAs对细胞生长、增殖、分化和凋亡等都具有重要的调节作用。前期研究表明，miR-146b是一段高度保守的miRNA，在恒河猴胚胎干细胞向神经元样细胞分化过程中显著高表达。
诱导多能干细胞：2006年，山中伸弥利用病毒载体将4个转录因子(Oct4，Sox2，Klf4和c-Myc)转入小鼠成纤维细胞中，使其重编程为类似胚胎干细胞的一种细胞类型。这类细胞在形态、基因和蛋白表达、表观遗传修饰、类胚体、畸胎瘤生成能力、分化能力等方面都与胚胎干细胞相似。由于诱导多能干细胞获得的方法相对简单和稳定，不需要使用卵细胞或者胚胎，在技术和伦理上都更具有优势，其在发病机制研究、新药筛选以及临床疾病治疗中都具有巨大的应用价值。

 

摘要
背景：如何促进诱导多能干细胞向神经元样细胞分化对于神经损伤修复和神经再生具有重要意义。
目的：探讨miR-146b在诱导多能干细胞分化过程中的表达水平，及其对诱导多能干细胞向神经元样细胞分化的影响。
方法：小鼠诱导多能干细胞向神经元样细胞分化7 d后，利用实时荧光定量PCR方法检测多能干细胞特异蛋白和miR-146b的表达；通过过表达miR-146b慢病毒感染诱导多能干细胞，再诱导其向神经元样细胞分化，利用细胞免疫荧光染色法和荧光定量PCR检测干细胞特异蛋白和神经外胚层特异蛋白的表达，以及神经细胞标记分子Tuj-1的表达。
结果与结论：①与未分化小鼠诱导多能干细胞相比，向神经元样细胞分化7 d后，干细胞特异表达蛋白Nanog、Oct4和Sox2表达显著下调，而miR-146b表达显著升高；②诱导多能干细胞过表达miR-146b后，Oct4表达显著下降，Nanog和Sox2表达未发生显著变化；同时，在神经分化过程中，过表达miR-146b诱导多能干细胞Nanog、Oct4和Sox2的表达显著下调，神经外胚层特异蛋白Nestin、Sox1和Pax6的表达显著上调；③神经分化培养18 d后，miR-146b显著增加了Tuj-1阳性细胞数；④结果表明，miR-146b可以促进诱导多能干细胞向神经元样细胞方向分化。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-2917-523X(李强)

关键词: miR-146b, 诱导多能干细胞, 神经元样细胞, 诱导分化, 干细胞

Abstract:

BACKGROUND: To facilitate the neuronal differentiation of induced pluripotent stem cells (iPSCs) is of great benefit for the repair of nerve injury and nerve regeneration.
OBJECTIVE: To investigate the expression of miR-146b in iPSCs differentiation, and its effects on the neural differentiation of iPSCs.
METHODS: After 7-day neural induction of mouse iPSCs, the expressions of miR-146b and pluripotent stem cell markers were detected by real-time PCR. Then, we generated miR-146b overexpressing iPSCs followed by the neural induction. The expression of stem cell and neuroectoderm markers were examined by cell immunofluorescence and real-time PCR respectively. Meanwhile, the expression of Tuj-1, a nerve cell marker, was also detected.
RESULTS AND CONCLUSION: Compared with the normal iPSCs, the expressions of pluripotent stem cell markers Nanog, Oct4 and Sox2 were significantly down-regulated in iPSCs undergoing 7-day neural induction, while the level of miR-146b was increased obviously. In miR-146b-overexpressing iPSCs, we found that the expression of Oct4 was substantially decreased, while there were no statistical changes in the expression of Nanog and Sox2. Meanwhile, during the neural differentiation, miR-146b overexpression significantly down-regulated the expression of Nanog, Oct4 and Sox2, and up-regulated the expression of neuroectoderm markers, Nestin, Sox1 and Pax6. After 18 days of neural induction, the miR-146b overexpression significantly increased the number of Tuj-1-positive cells. Taken together, miR-146b plays a vital role in facilitating the differentiation of iPSCs into neuron-like cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Induced Pluripotent Stem Cells, MicroRNAs, Neurons, Cell Differentiation, Tissue Engineering

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