参枣健脑口服液促进神经干细胞向神经元分化的作用机制研究

Study on the Mechanism of Shenzao Jiannao Oral Liquid Promoting the Differentiation of Neural Stem Cells into Neurons

目的探究参枣健脑口服液(SZJN)对神经干细胞(NSCs)分化的影响及其作用机制。方法原代培养海马NSCs,构建阿尔茨海默病(AD)细胞模型。CCK-8法检测AD细胞模型(APP-NSCs)细胞活力,微量酶标法检测乳酸脱氢酶释放量,神经球形成实验检测APP-NSCs细胞增殖能力,免疫荧光染色检测APP-NSCs细胞分化情况,NSCs迁移实验检测神经元迁移距离,Western blot法检测Wnt/β-catenin通路关键蛋白的表达情况。结果SZJN能够有效提高APP-NSCs细胞活力,减轻APP-NSCs损伤,增加神经球直径和神经球数量,促进NSCs和APP-NSCs向神经元方向分化,增加NSCs和APP-NSCs神经元迁移距离,提高Wnt-3a和β-catenin蛋白表达量,促进GSK-3β磷酸化。结论SZJN可通过激活Wnt/β-catenin信号通路促进NSCs向神经元分化。

Objective To study the effect of Shenzao Jiannao oral liquid(SZJN)on the differentiation of neural stem cells(NSCs),and explore the underlying mechanism.Methods Primary culture of hippocampal neural stem cells to construct a model of Alzheimer’s disease cells.The activity of APP-NSCs cells is detected by CCK-8 assays,the level of lactic dehydrogenase in the culture is tested to assess the damage of APP-NSCs cell,immunofluorescence is used to detect the differentiation of APP-NSCs,the migration distances of neurons are measured to evaluate the migration of NSCs,Western blot analysis is used to detect the expression of key proteins in the Wnt/β-catenin pathway.Results SZJN can effectively improve the cell viability of APP-NSCs,alleviate the injury of APP-NSCs,increase the diameter and number of neurospheres,promote the neuronal differentiation of NSCs and APP-NSCs,and increase the migration distance of NSCs and APP-NSCs.Moreover,SZJN can also increase the protein levels of Wnt-3a andβ-catenin in neurons derived from APP-NSCs,as well as promote the phosphorylation of GSK-3β.Conclusion SZJN can promote the proliferation of APP-NSCs,as well as the neuronal differentiation of APP-NSCs by activating the Wnt/β-catenin signaling pathway.