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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。
In order to explore the role of ginsenoside Rg1 in inducing the differentiation of hippocampal neural stem cells (NSCs) in neonatal rats, the cells were cultured in serum-free and monoclonal culture methods. The morphology of NSCs and the differentiated neurons were observed by immunofluorescence staining , The morphology of glial cells, and image analysis of the number of cells differentiated into hippocampal NSCs. The experiment was divided into 4 groups: 5% fetal bovine serum group, 5% fetal bovine serum +30 mg / kg ginsenoside Rg1 group, 5% fetal bovine serum +15 mg / kg ginsenoside Rg1 group and 5% fetal bovine serum +10 mg / kg ginsenoside Rg1 group. The results showed that hippocampal NSCs could form nestin and BrdU positive cells in serum-free culture. After induced differentiation, immunofluorescence staining revealed that the expression of β-Ⅲ tubulin (Tuj -1) and vimentin positive cells. The results of image analysis showed that the NSCs differentiated into hippocampal neurons in the group of 15 mg / kg ginsenoside Rg1 were the highest. The results suggest that under certain concentration of ginsenoside Rg1, the number of NSCs differentiated into neurons in hippocampus increased, which provided experimental evidence for further study on the effect of ginsenoside Rg1 on the differentiation of NSCs.