人胚胎干细胞神经分化中VEGF作用的分子机制研究  

Molecular mechanism of VEGF on neural differentiation of human embryonic stem in vitro

作　　者：焦淑洁[1,2] 许慧芳[1] 张苏明[1] 许杰[1] 湛延强[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030 [2]郑州大学第一附属医院神经内科,郑州450052

出　　处：《中国实用神经疾病杂志》2011年第19期4-6,共3页Chinese Journal of Practical Nervous Diseases

摘　　要：目的观察血管内皮生长因子(vascular endothelial growth factor,VEGF)对人胚胎干细胞分化为神经元的促进作用是否与p38MAPK信号通路相关。方法人胚胎干细胞经拟胚体向神经元分化,分为3组:A组:常规诱导组;B组:常规诱导+VEGF(10ng/mL)作用组;C组:常规诱导+p38MAPK抑制剂预处理+VEGF(10ng/mL)作用组。免疫荧光法检测计算各组不同阶段细胞阳性率,半定量RT-PCR观察各组神经干细胞p38αmRNA表达,MTT法检测3组神经干细胞增殖速率。结果免疫荧光法检测,B、C 2组产生神经干细胞的阳性率明显高于A组,差异有统计学意义(P<0.01),B、C 2组比较差异无统计学意义(P>0.05);B组进一步分化为神经元的阳性率明显高于A、C组,差异有统计学意义(P<0.01),A、C 2组比较差异无统计学意义(P>0.05);半定量RT-PCR观察B组神经干细胞p38αmRNA表达明显高于A、C 2组;MTT法检测与半定量RT-PCR结果相似。结论人胚胎干细胞体外分化过程中,血管内皮生长因子促进神经干细胞分化的作用与p38MAPK通路无关,而VEGF通过激活p38MAPK信号通路,促进神经干细胞进一部分化为神经元。Objective To study the mechanism of VEGF on the neural differentiation of hESCs in vitro. Methods hESCs were cultured and differentiated into neurons through the embryonic body. Before the VEGF （10 ng/mL） was added, p38 MAPK inhibitor was given at every stage of induction for 1 hour. The markers of every stage were detected and the percentages of the imunostaining positive cells were counted through immunofluoresce group were detected by RT-PCR. Results The applications of p38 MAPK nce. The expressions of p38 MAPK mRNA in each inhibitor produced the similar number of neural stem ceils with the VEGF group; while the neurons of p38 MAPK inhibitor pretreatment group and conventional induction group were similar. RT-PCR analysis showed that the expression of p38 mRNA in the VEGF treated group was higher than p38 MAPK inhibitor preconditioning group and the conventional group. Conclusion In the differentiation of human neural stem cells into neurons, VEGF can promote neural stem cells to differentiate into more neurons via p38MAPK way. p38 MAPK inhibitors can block the role of VEGF in this process.

关 键 词：人胚胎干细胞 神经分化 血管内皮生长因子 P38MAPK信号通路 

分 类 号：R34[医药卫生—基础医学]