黄芪多糖对C17.2神经干细胞定向分化的调控作用  

Experimental Study on Regulating Directional Differentiation of C17.2 Neural Stem Cells by Astragalus Polysaccharide in Vitro

作　　者：叶妮 马金昀 程晓东 Ye Ni;Ma Jinyun;Cheng Xiaodong(Institute of Clinical Immunology,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院临床免疫研究所

出　　处：《世界中医药》2019年第10期2598-2602,2608,共6页World Chinese Medicine

基　　金：国家自然科学基金项目(81673669,81703782);上海市交叉学科“中医临床免疫学”建设项目(30304113598);上海中医药大学高峰学科建设项目(30304114323);上海中医药大学研究生创新培育项目(JX61.02.03.55)

摘　　要：目的:观察黄芪多糖(APS)对C17.2神经干细胞体外定向分化的调控作用。方法:建立C17.2神经干细胞的体外培养体系及分化模型,设置APS干预组及PBS对照组。诱导分化4 d后,通过细胞免疫荧光染色的方法检测2组细胞中神经干细胞的标志性蛋白巢蛋白(Nestin)、星形胶质细胞的标志性蛋白胶质纤维酸性蛋白(GFAP)、少突胶质细胞的标志性蛋白髓鞘碱性蛋白(MBP)及神经元的标志性蛋白神经元特异性核心抗原(NeuN)的表达水平。结果:与PBS对照组比较,APS干预组细胞的Nestin及GFAP蛋白表达水平明显下调,MBP及NeuN蛋白表达水平明显上调,差异有统计学意义(P<0.05)。APS下调了分化模型中的细胞Nestin蛋白的表达水平;APS下调了GFAP蛋白的表达水平,上调了MBP及NeuN蛋白的表达水平。结论:APS可抑制C17.2神经干细胞向星形胶质细胞的定向分化,促进向少突胶质细胞及神经元的定向分化,APS可抑制C17.2神经干细胞的干性维持,促进其进入分化状态,有可能成为治疗神经退行性疾病的潜在药物。Objective:To observe regulatory effects of Astragalus Polysaccharide(APS)on directional differentiation of C17.2 neural stem cells in vitro.Methods:Cultivation system and differentiation model of C17.2 neural stem cells were established in vitro.APS intervention group and PBS control group were set up.Immunofluorescence staining was used to detect expression levels of Nestin,GFAP,MBP and NeuN,the simbolic marker proteins of neural stem cells,astrocytes,oligodendrocytes and neurons,so as to explore the regulatory effects of APS on directional differentiation of C17.2 neural stem cells.Results:Compared with the PBS control group,the expression levels of Nestin and GFAP proteins in the APS intervention group were down-regulated while the expression levels of MBP and NeuN proteins were up-regulated.The differences were statistically significant(P<0.05).APS down-regulated the expression level of Nestin protein in the differentiation model,which meant it could inhibit the stemness maintenance of C17.2 neural stem cells and lead them to enter the differentiation state.APS down-regulated the expression level of GFAP protein,and up-regulated the expression levels of MBP and NeuN proteins,which meant it could promote C17.2 neural stem cells to differentiate into oligodendrocytes and neurons directionally,and inhibit them to differentiate into astrocytes.Conclusion:APS can effectively regulate the directional differentiation of C17.2 neural stem cells in vitro,which suggests that it might be a potential drug for the treatment of neurodegenerative diseases.

关 键 词：神经干细胞 定向分化 黄芪多糖 少突胶质细胞 神经元 星形胶质细胞 神经退行性疾病 体外研究 

分 类 号：R284[医药卫生—中药学]