中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (6): 992-995.doi: 10.3969/j.issn.1673-8225.2010.06.008

• 干细胞培养与分化 • 上一篇    下一篇

体外缺氧条件下新生鼠神经干细胞pERK1/2表达与叶酸的影响

杨  阳，黄国伟，张绪梅，赵  琳，尤清菊，刘佳杰   

  1. 天津医科大学公共卫生学院，天津市  300070
  • 出版日期:2010-02-05 发布日期:2010-02-05
  • 通讯作者: 黄国伟，博士，教授，博士生导师，天津医科大学公共卫生学院，天津市 300070 guoweihuang@yahoo.com.cn
  • 作者简介:杨 阳，女，1981年生，辽宁省沈阳市人，汉族，天津医科大学在读硕士，主要从事营养与慢性病方面的研究。 yueyangzhen@yahoo.com.cn
  • 基金资助:

    国家自然科学基金资助项目（30771797），课题名称“叶酸对神经干细胞增殖分化作用的蛋白质组学的研究”；
    天津医科大学科学基金资助项目（2008Ky12），课题名称“同型半胱氨酸对大鼠学习记忆及ERK信号通路的影响”。

Effects of folic acid on pERK1/2 expression of neural stem cells from neonatal rats under hypoxia condition in vitro

Yang Yang, Huang Guo-wei, Zhang Xu-mei, Zhao Lin, You Qing-ju, Liu Jia-jie   

  1. College of Public Health, Tianjin Medical University, Tianjin   300070, China
  • Online:2010-02-05 Published:2010-02-05
  • Contact: Huang Guo-wei, Doctor, Professor, Doctoral supervisor, College of Public Health, Tianjin Medical University, Tianjin 300070, China guoweihuang@yahoo.com.cn
  • About author:Yang Yang, Studying for master’s degree, College of Public Health, Tianjin Medical University, Tianjin 300070, China yueyangzhen@yahoo.com.cn
  • Supported by:

    the National Natural Science Foundation of China, No.30771797*;
    the Science Foundation of Tianjin Medical University, No.2008Ky12*

PDF

400

摘要：

背景：课题组前期实验已经证实，神经干细胞在正常培养条件下，叶酸可通过丝裂原活化蛋白激酶通路激活ERK1/2的磷酸化，进而促进神经干细胞的增殖。

目的：探讨叶酸在体外缺氧条件下对神经干细胞外信号调节蛋白激酶pERK1/2表达的影响。

方法：采用无血清培养法体外分离培养新生鼠神经干细胞，以1×108 L-1接种于培养瓶，设立4组，除正常对照组外，缺氧模型组、叶酸缺乏组、叶酸添加组细胞均于第3天放入自制缺氧装置，37 ℃恒温箱中缺氧培养6 h，4组的叶酸含量分别为4 mg/L，4 mg/L，0.65 mg/L，8 mg/L。收集增殖6 d的细胞，锥虫蓝计数细胞密度，RT-PCR法检测pERK1/2 mRNA的表达，Western blot法检测pERK1/2蛋白的表达。

结果与结论：与正常对照组比较，缺氧模型组神经干细胞增殖能力、pERK1/2 mRNA及蛋白的表达均明显降低。与缺氧模型组比较，叶酸添加组能促进缺氧条件下神经干细胞增殖以及pERK1/2 mRNA、蛋白的表达，而叶酸缺乏组则抑制缺氧条件下神经干细胞的增殖以及pERK1/2 mRNA、蛋白的表达，各组间比较差异有显著性意义(P < 0.001)。证实添加叶酸后可激活ERK1/2磷酸化，进而促进缺氧条件下神经干细胞的增殖。

关键词: 叶酸, 缺氧, 增殖, pERK1/2, 神经干细胞

Abstract:

BACKGROUND: Previous studies have verified that under normal culture of neural stem cells (NSCs), folic acid can accelerate proliferation of NSCs by phosphorylation of mitogen activated protein kinase path activation ERK1/2.

OBJECTIVE: To investigate the effect of folic acid on NSC extracellular signal regulatory protein kinase pERK1/2 under hypoxic condition.

METHODS: NSCs from Neonatal rats were cultured in vitro by serum-free culture method, and incubated in a flask at 1×108/L. Except normal control group, self-made hypoxia equipment was used in the hypoxia model, folic acid deficiency and folic acid supplemented groups at day 3. At 37 ℃, hypoxia culture was conducted in the thermostat for 6 hours. The contents of folic acid were 4 mg/L, 4 mg/L, 0.65 mg/L, 8 mg/L in the four groups. Cells following 6 days were collected to count the density using trypan blue. RT-PCR was utilized to detect pERK1/2 mRNA expression. Western blot assay was employed to determine pERK1/2 protein expression.

RESULTS AND CONCLUSION: Compared with the normal control group, the proliferation of NSCs and the expression of ERK1/2mRNA and pERK1/2 protein were decreased significantly in the hypoxia model group. Compared with the hypoxia model group, the proliferation of NSCs and the expression of ERK1/2mRNA and pERK1/2 protein were increased in the folic acid supplemented group, whereas decreased in the folic acid deficiency group. There were significant differences among groups (P < 0.001). Above-described results verified that folic acid supplementation can activate ERK1/2 phosphorylatin and accelerate proliferation of NSCs under hypoxia condition.

中图分类号: