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双向电泳分析叶酸对神经干细胞蛋白表达的影响

刘欢1,黄国伟1,何冰2,杨阳1,赵琳1   

  1. 1. 天津医科大学
    2. 武警医学院天津市职业与环境危害生物标志物重点实验室
  • 收稿日期:2011-01-05 修回日期:2011-02-24 出版日期:2011-07-15 发布日期:2011-07-15
  • 通讯作者: 刘欢

Two-dimensional Gel Electrophoresis Analysis for Effect of Folate on Protein Expression in neural stem cells

  • Received:2011-01-05 Revised:2011-02-24 Published:2011-07-15 Online:2011-07-15

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摘要： 目的：比较叶酸缺乏和叶酸补充干预下胚胎大鼠神经干细胞（Neural Stem Cells，NSCs）蛋白质表达的差异。方法：采用无血清体外细胞培养方法，培养胚胎大鼠神经干细胞并进行鉴定，采用叶酸缺乏培养基培养NSCs，根据叶酸添加终浓度将细胞分为对照组（Control），叶酸终浓度为4μg/mL；叶酸缺乏组（Folate-D），叶酸浓度为0.6μg/mL；叶酸补充组（Folate-L），叶酸终浓度为8μg/ml，应用双向电泳（2-DE）技术分析各组蛋白质表达的差异情况。结果：建立了稳定可重复的NSCs2-DE图谱，与Control组相比，Folate-D组2个蛋白点表达量降低，4个蛋白点表达量升高；Folate-L组2个蛋白点表达量降低，1个蛋白点表达量升高。结论：叶酸缺乏或补充影响NSCs蛋白表达，差异蛋白分离为进一步研究叶酸对NSCs增殖分化调控机制奠定良好基础。

关键词: 叶酸, 神经干细胞, 双向电泳, 蛋白质组学

Abstract: Objective: To analyze the differentially expressed proteins of embryonic rat neural stem cells (NSCs) under folate deficiency or supplementation. Methods: Embryonic rat NSCs were cultured in serum-free suspension culture. Cells were identified by their expression of immunoreactive nestin. Individual cultures were assigned to one of 3 treatment groups that differed according to the concentration of folate in the medium: Control (baseline folate concentration of 4μg/mL), folate deficiency group (Folate-D, folate concentration of 0.6μg/mL) and folate supplementation (4μg/mL above baseline, Folate-L). Protein expressions in 3 groups were separated by two-dimensional electrophoresis (2-DE). Results: The stable and repeatable 2-DE maps of NSCs were successfully established. Compared with Control group, 2 proteins in Folate-D group were lowly expressed, 4 proteins were highly expressed. 2 protein spots were lowly expressed and 1 protein was highly expressed in Folate-L group. Conclusion: Folate deficiency or supplement may impact NSCs protein expression. The isolation of differentially expressed proteins will be useful for exploring the mechanism of folate on proliferation and differentiation of NSCs.

Key words: folate, neural stem cells, two-dimensional, proteome