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目的探讨音猬因子(SHH)对神经干细胞(NSCs)增殖与分化的影响。方法从大鼠胚胎端脑皮质及海马分离培养的NSCs分为SHH组、RA组和PBS组。分别用SHH、维甲酸(RA)及PBS条件培养液处理后,通过免疫细胞化学等方法检测NSCs的增殖及分化情况。结果体外培养的NSCs能快速增殖并形成神经球,其中的细胞均表达NSCs特异性标志物nestin。BrdU作用于NSCs3h后,SHH组的BrdU阳性率明显高于RA组和PBS组,而RA组和PBS组之间无显著差异。当NSCs在分化条件培养液中培养7d后,SHH组和RA组中的βⅢ-tubulin阳性率及Rip阳性率显著高于PBS组,SHH组又显著高于RA组和PBS组。结论SHH可以促进NSCs增殖及其向神经元和少突胶质细胞的分化。
Objective To investigate the effect of sonic hedgehog (SHH) on the proliferation and differentiation of neural stem cells (NSCs). Methods NSCs isolated from rat embryonic cortex and hippocampus were divided into SHH group, RA group and PBS group. The proliferation and differentiation of NSCs were detected by immunocytochemistry after treated with SHH, RA and PBS respectively. Results The NSCs cultured in vitro could rapidly proliferate and form neurospheres. All of them expressed nestin, a specific marker of NSCs. After BrdU treatment for 3h, the positive rate of BrdU in SHH group was significantly higher than that in RA group and PBS group, but there was no significant difference between RA group and PBS group. When NSCs were cultured in differentiation medium for 7 days, the positive rate of βⅢ-tubulin and the positive rate of Rip in SHH group and RA group were significantly higher than those in PBS group, and were significantly higher in SHH group than those in RA group and PBS group. Conclusion SHH can promote the proliferation of NSCs and their differentiation into neurons and oligodendrocytes.