上海交通大学学报（医学版）

›› 2009, Vol. 29 ›› Issue (10): 1191-.

• 论著（基础研究） • 上一篇    下一篇

大鼠神经干细胞中PI3-K/AKT信号转导通路的研究

赵 宇1, 谢 鹏1, 朱晓峰2   

  1. 1. 重庆医科大学 附属第一医院神经内科, 重庆 400016；2. 佳木斯大学神经科学研究所, 佳木斯 154007
  • 出版日期:2009-10-25 发布日期:2009-10-26
  • 作者简介:赵 宇（1973—）, 女, 副主任医师, 博士生；电子信箱: wangpang0916@yahoo.com.cn；现在哈尔滨医科大学附属第四医院神经内科工作。
  • 基金资助:

    高等学校博士学科点专项科研基金（20050631012）

Effects of PI3-K/AKT signal transduction pathway in rat neural stem cells

ZHAO Yu1, XIE Peng1, ZHU Xiao-feng2   

  1. 1. Department of Neurology, The First Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China；2. Neuroscience Research Institute, Jiamusi University, Jiamusi 154007, China
  • Online:2009-10-25 Published:2009-10-26
  • Supported by:

    University Doctoral Subject Research Foundation, 20050631012

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摘要：

目的 应用PI3-K特异性阻滞剂LY294002孵育大鼠神经干细胞（NSCs），探讨PI3-K/AKT信号转导通路在NSCs中的作用。方法 体外分离、培养E15～16 胎鼠NSCs，应用不同浓度（0~40 μmol/L）的LY294002孵育NSCs后进行细胞存活(WST-8检测)、增殖(BrdU免疫组织化学鉴定)、分化(β Tubulin-Ⅲ免疫组织化学鉴定)和AKT磷酸化蛋白水平(Western Blotting)的检测。结果 LY294002对NSCs的存活、增殖和分化的影响呈剂量依赖性。在LY294002较高浓度（25、30、35、40 μmol/L）时，NSCs的存活率明显下降（P<0.05）；LY294002为30、35、40 μmol/L时，BrdU阳性细胞数明显少于正常对照组（LY294002 0 μmol/L）（P<0.05）；LY294002为35、40 μmol/L时，β Tubulin-Ⅲ的阳性细胞数明显少于正常对照组（P<0.05）。LY294002 阻断AKT表达的作用呈剂量依赖性。结论 PI3-K/AKT信号转导通路对大鼠NSCs的存活、增殖、分化起重要作用。

关键词: 磷脂酰肌醇3激酶, 蛋白激酶B, 神经干细胞, 大鼠

Abstract:

Objective To incubate rat neural stem cells (NSCs) with selection-specific inhibitor (LY294002) of PI3-K, and explore the effects of PI3-K/AKT signal transduction pathway in NSCs. Methods NSCs derived from E15-16 rats were isolated and cultured. After incubation of NSCs with different concentrations of LY294002 (0 to 40 μmol/L), cell survival (WTS-8 assay), proliferation (BrdU immunohistochemical identification), differentiation (β Tubulin-Ⅲ immunohistochemical identification) and AKP phosphorylated protein (Western blotting) were detected. Results The effects of LY294002 on the survival, proliferation and differentiation of NSCs was concentration-dependent. The survival of the NSCs decreased significantly with a higher concentrations of LY294002 (25, 30, 35, 40 μmol/L)(P<0.05). The numbers of BrdU positive cells with LY294002 of 30, 35 and 40 μmol/L were significantly smaller than that of the control (LY294002 0 μmol/L)(P<0.05). The numbers of β Tubulin-Ⅲ positive cells with LY294002 of 35 and 40 μmol/L were significantly smaller than that of the control (P<0.05). The expression of AKT was blocked by LY294002 in a concentration-dependent manner. Conclusion PI3-K/AKT signal transduction pathway plays a vital role in the survival, proliferation and differentiation of NSCs.

Key words: phosphatidylinositol 3 kinase, protein kinase B, neural stem cells, rat

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