人参皂苷Rg1诱导大鼠海马神经干细胞分化的实验研究

Experimental study of ginsenoside Rg1 on the differentiation of rat hippocampus neural stem cells

为探讨人参皂苷Rg1诱导新生大鼠海马神经干细胞(NSCs)分化的作用,本实验采用无血清和单克隆培养方法进行细胞培养,应用免疫荧光染色法观察NSCs的形态及其分化而来的神经元、胶质细胞的形态,并经图像扫描分析仪分析海马NSCs分化的细胞数量。实验分4组:5%胎牛血清组、5%胎牛血清+30 mg/kg人参皂苷Rg1组、5%胎牛血清+15 mg/kg人参皂苷Rg1组和5%胎牛血清+10 mg/kg人参皂苷Rg1组。结果显示:海马NSCs在无血清培养下,可形成巢蛋白(nestin)和5-溴脱氧尿苷(BrdU)阳性细胞球;在诱导分化后,免疫荧光染色可见β-Ⅲ型微管蛋白(Tuj-1)与波形蛋白(vimentin)阳性细胞;图像扫描分析结果显示15 mg/kg人参皂苷Rg1干预组海马NSCs向神经元的分化比例最高。此结果提示在一定浓度人参皂苷Rg1的作用下,海马NSCs向神经元分化的数量增多,为进一步研究人参皂苷Rg1对NSCs分化的影响提供了实验依据。

To observe the effects of ginsenoside Rgl on the differentiation of neonate rat hippocampus neural stem cells （ NSCs）, the neo- nate rat hippocampus stem cells were cultured by serum free medium and single cell cloning technique in this study. Morphous of NSCs, neurons and astrocytes was observed by immunofluorescence staining method, and the number of positive cells from hippocampus NSCs was counted by image scanning analysator. The cultured NSCs were divided into four groups： 5% fetal bovine serum （FBS） group, 5% FBS ＋30 mg/kg ginsenoside Rgl group, 5% FBS ＋ 15 mg/kg ginsenoside Rgl group, and 5% FBS ＋ 10 mg/kg gir/senoside Rgl group. The results showed that the NSCs of hippocampus cultured by serum free medium could form nestin and BrdU positive cellular spheres. The stem cells of hippocampus can differentiate into Tuj-1 and vimentin positive ceils by differently inducing. The ratio of hippocampal NSCs differentiated into neurons was maximum in the 15 mg/kg consistency of ginsenoside Rgl group by image scanning analysator. The present results suggest that the number of hippocampal NSCs differentiated into neurons increases with effect of a certain concentration ginsenoside Rgl. It also provides the experimental basis for further research about the effect of ginsenoside Rgl on the differentiation of NSCs.