【摘要】 目的:探讨银杏内酯B(GKB)对大鼠中脑神经干细胞(NSCs)体外诱导分化为多巴胺能神经元的作用。方法:采用无血清培养技术从大鼠胚胎中脑组织中分离培养出神经干细胞克隆球,然后分为2组诱导分化,对照组分化培养液中仅含10%FBS,银杏内酯B组在对照组分化培养液的基础上加入终浓度为40μg/ml的银杏内酯B。2周后免疫印迹检测分化的细胞中多巴胺能神经元发育和分化调节因子Nurr1蛋白的表达情况;免疫荧光检测分化神经元中酪氨酸羟化酶(TH)标记的多巴胺能神经元数量和分化情况。结果:对照组Nurr1蛋白相对表达量很低,仅有极少量的神经干细胞分化为不成熟的多巴胺能神经元;银杏内酯B组Nurr1表达显著升高,分化的多巴胺能神经元较多,且细胞形态较成熟,突起较丰富。结论:银杏内酯B具有诱导中脑NSCs向多巴胺能神经元分化的作用,其作用机制可能与上调Nurr1表达有关。更多还原

【Abstract】 Objective:To explore effect of ginkgolide B(GKB) on inducing neural stem cells(NSCs) derived from rat midbrain to differentiate into dopaminergic neurons in vitro.Methods:Neurospheres were obtained from rat fetal midbrain by serumfree culturing,and were divided into 2 groups.In the control group,the differentiation medium only contained 10%FBS.In GKB group,the medium contained 10%FBS and 40 μg/ml GKB.After 2 weeks,the expression of Nurr1 protein in differentiated cells was detected by Western blotting,and the tyrosine hydroxylase(TH) positive cells were detected by immunofluorescence.Results:In the control group,the relative level of Nurr1 was low,and there were few TH positive neurons with poorly developed morphology.While the expression of Nurr1 was increased significantly in GKB group,more dopaminergic neurons were produced with fully differentiated neuronal shapes and extensive neurite fibres.Conclusion:Ginkgolide B has an obvious effect on inducing NSCs derived from fetal mescenphalon to dopaminergic neurons,and the possible mechanism may be associated with the up-regulation of Nurr1.更多还原