银杏内酯B对神经干细胞分化的影响及其机制研究

Mechanisms and Effect of Ginkgolide B on the Differentiation of Neuron Stem Cells

目的观察银杏内酯B对体外培养的神经干细胞(NSCs)分化的影响。方法用含不同浓度银杏内酯B(20mg/L、40mg/L、60mg/L)的分化培养基培养NSCs3d和7d,测量细胞突起长度和胞体面积,用免疫荧光检测神经丝蛋白-200(NF-200)、胶质纤维酸性蛋白(GFAP)及少突胶质细胞特异性蛋白(CC-1)、细胞因子信号抑制因子2(SOCS2)、DNA结合抑制因子2(Id2)的表达。结果银杏内酯B组细胞突起增长,胞体增大;NF阳性神经元样细胞百分率增加,GFAP阳性星形胶质样细胞百分率随银杏内酯B浓度增加而增加;SOCS2阳性细胞百分率(35.46%)较对照组(22.17%)明显增加(P<0.01),银杏内酯B组Id2阳性细胞百分率(51.52%)较对照组(66.24%)明显减少(P<0.01)。结论银杏内酯B可促进分化细胞成熟,促进NSCs向神经元分化;星形胶质细胞百分率的增高与银杏内酯B呈剂量依赖关系。

Objective To observe the effect of Ginkgolide B of various consistency on the differentiation of neuron stem cells （NSCs）. Methods NSCs were cultured in differentiation medium containing Ginkgolide B of various consistency for 3 and 7 days, the neurites length and cell body area were measured by inverted phase-contrast micrograph, then neurofilament-200 （NF-200）, glial fibrillary acidic protein （GFAP）, adenomatus polyposis coli （CC-1） expression were detected and counted by fluorescence microscope. The suppressor of cytokine signaling-2 （SOCS2）, inhibitor of DNA binding-2 （Id2） were alsoimmunostained. The percentage of positive cells were counted respectively. Results The neurites length and cell body area in Ginkgolide B groups were obviously larger than that in the control group. The percentage of NF, GFAP positive cells in Ginkgolide B groups increased with dosage increasing of Ginkgolide B. Compared with the normal control group, the percentage of SOCS2 positive cells increased significantly （ P〈0. 01） and the percentage of Id2 positive cells decreased significantly （ P〈0.01） in Ginkgolide B groups. Conclusion Ginkgolide B can promote NSCs to differentiate into neuron and astrocyte, the percentage of astrocyte is increased with a dosage-dependent relationship with Ginkgolide B.