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目的:采用基因芯片技术筛选出人参皂苷Rg1促进NSCs分化的主要分子靶点。方法:通过基因芯片技术,观察Rg1诱导人神经干细胞(neural stem cells,NSCs)向神经元分化7 d时靶基因表达,通过数据演算筛选出Rg1促进NSCs分化的最主要的靶基因和信号转导途径,然后采用Western blot和免疫组化的方法对其中的ERK信号分子进行验证。结果:在Rg1诱导NSCs分化第7天时,获得差异基因675个,其中显著上调的基因255个,显著下调的基因420个;MAPK(丝裂原活化蛋白激酶)通路中的ERK1/2(细胞外信号调节蛋白激酶)信号分子与NSCs分化直接相关。经Western blot和免疫组化证实,在Rg1诱导NSCs分化中,ERK1/2蛋白明显上调,磷酸化水平也明显增强,此作用能够被PD98059(ERK1/2阻断剂)所阻断,同时PD98059也可以明显阻断NSCs的分化。结论:ERK1/2是人参皂苷Rg1促进NSCs分化的重要分子靶点。基因芯片筛选出的差异表达基因可能为研究Rg1促进NSCs分化的分子机制提供线索。
OBJECTIVE: To screen out the main molecular targets of ginsenoside Rg1 for differentiation of NSCs by gene chip technology. Methods: The gene expression of Rg1 on neural stem cells (NSCs) induced to neurons for 7 days was observed by gene chip technique. The most important target genes and signal transduction of Rg1 in promoting differentiation of NSCs were screened by data calculation Pathway, and then use Western blot and immunohistochemical methods to verify the ERK signaling molecules. RESULTS: On day 7 of Rg1-induced NSCs differentiation, 675 genes were differentially expressed, including 255 significantly up-regulated genes and 420 significantly down-regulated genes. ERK1 / 2 (extracellularly) in the mitogen-activated protein kinase Signal-regulated protein kinase) signaling molecules are directly related to the differentiation of NSCs. Western blot and immunohistochemistry showed that ERK1 / 2 protein was significantly up-regulated and phosphorylation was significantly increased in Rg1-induced NSCs, which was blocked by PD98059 (ERK1 / 2 blocker) and PD98059 Can significantly block the differentiation of NSCs. Conclusion: ERK1 / 2 is an important molecular target of ginsenoside Rg1 in promoting the differentiation of NSCs. The differentially expressed genes screened by gene chip may provide clues for studying the molecular mechanism of Rg1 promoting NSCs differentiation.