上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (12): 1715-.doi: 10.3969/j.issn.1674-8115.2011.12.012

• 论著（基础研究） • 上一篇    下一篇

GDNF在精原干细胞中对H19和A-myb表达的调控研究

查巍巍1, 曹静萍2, 张 迪1, 冯立新1, 程金科1   

  1. 1.上海交通大学 医学院干细胞研究室, 上海 200025； 2.中国科学院上海生命科学研究院/上海交通大学医学院健康科学研究所, 上海 200025
  • 出版日期:2011-12-28 发布日期:2012-01-04
  • 通讯作者: 冯立新, 电子信箱: fenglx66@yahoo.com。
  • 作者简介:查巍巍（1981—）, 女, 硕士生；电子信箱: zhaweimm@gmail.com。

Role of GDNF in regulation of expression of H19 and A-myb in spermatogonial stem cells

ZHA Wei-wei1, CAO Jing-ping2, ZHANG Di1, FENG Li-xin1, CHENG Jin-ke1   

  1. 1.Laboratory for Stem Cell Research, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China；2.Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2011-12-28 Published:2012-01-04

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摘要：

目的 通过研究胶质细胞神经源性营养因子（GDNF）对H19和A-myb表达的调控，探讨GDNF调节精原干细胞自我更新的分子机制。方法 采用混合酶消化和差速贴壁法从出生后6 d的小鼠睾丸中分离、鉴定并培养精原干细胞。用GDNF分别刺激精原干细胞和经磷酸肌醇3激酶/蛋白激酶B(PI3K/Akt)通路抑制剂LY294002预处理的精原干细胞后，采用RT-PCR技术检测细胞H19和A-myb的表达。结果 相关鉴定分析证实分离培养的细胞为精原干细胞。RT-PCR检测结果显示：GDNF刺激下的精原干细胞的H19和A-myb表达均显著上调，而经LY294002预处理的精原干细胞在GDNF刺激下其H19和A-myb的表达无明显变化。结论 在体外培养的小鼠精原干细胞中，GDNF通过PI3K/Akt 信号通路上调H19和A-myb的表达。

关键词: 胶质细胞源性神经营养因子, 精原干细胞, H19, A-myb, 磷酸肌醇3激酶/蛋白激酶B

Abstract:

Objective To investigate the regulation of expression of H19 and A-myb by glial cell line-derived neurotrophic factor (GDNF), and explore the molecular mechanism of self-renewal of spermatogonial stem cells regulated by GDNF. Methods Spermatogonial stem cells were isolated, identified and cultured from testes of 6-day-old mice with mixed enzyme digestion and differential plating procedure. Spermatogonial stem cells and spermatogonial stem cells pretreated with LY294002, inhibitor of phosphotylinosital 3 kinase/protein kinase B(PI3K/Akt), were stimulated with GDNF, and the expression of H19 and A-myb in spermatogonial stem cells was detected by RT-PCR. Results Spermatogonial stem cells were successfully isolated and cultured. The expression of H19 and A-myb was upregulated in spermatogonial stem cells after stimulation with GDNF, while there was no significant change in the expression of H19 and A-myb in spermatogonial stem cells after pretreatment with LY294002. Conclusion GDNF can upregulate the expression of H19 and A-myb via PI3K/AKT signal pathway in cultured mouse spermatogonial stem cells.

Key words: glial cell line-derived neurotrophic factor, spermatogonial stem cell, H19, A-myb, phosphotylinosital 3 kinase/protein kinase B