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目的观察胰岛素样生长因子-1(IGF-1)对体外培养神经干细胞(NSCs)向神经元分化的影响,并探讨PI3K/Akt信号通路与之的关系。方法取新生C57BL/6J小鼠海马组织,分离、培养NSCs,将细胞团吹散,以1×104个/mL密度接种于24孔板,分别加入终浓度100 ng/mL的IGF-1和50μmol/L的PI3K/Akt通路特异抑制剂LY294002,根据处理的情况,将细胞分为IGF-1组、正常对照组、LY组和IGF-1+LY组。βⅢTubulin免疫荧光细胞化学染色法检测NSCs分化为神经元的情况,DAPI染核计细胞总数;p-Akt免疫荧光染色检测各组细胞Akt磷酸化的情况;Western blotting法检测Akt和p-Akt蛋白的表达量,并对各组进行比较。结果 IGF-1组NSCs向神经元分化的比率显著高于对照组、LY组和IGF-1+LY组(P均<0.05);免疫荧光染色结果显示,IGF-1组Akt磷酸化水平显著高于对照组、LY组和IGF-1+LY组(P均<0.05);Western blotting结果显示,IGF-1促进Akt磷酸化,LY294002抑制Akt的磷酸化。结论 IGF-1诱导Akt的磷酸化,从而激活PI3K/Akt信号通路,促进NSCs向神经元分化。
Objective To observe the effects of insulin-like growth factor-1 (IGF-1) on the differentiation of neural stem cells (NSCs) into neurons in vitro and to explore the relationship between PI3K / Akt signaling pathway and neuronal differentiation. Methods The hippocampus of newborn C57BL / 6J mice were isolated, cultured, and NSCs were isolated. The cells were blown away and seeded into 24-well plates at a density of 1 × 10 4 / mL. IGF-1 and 50 μmol / L PI3K / Akt pathway specific inhibitor LY294002, according to the treatment, the cells were divided into IGF-1 group, normal control group, LY group and IGF-1 + LY group. βⅢTubulin immunofluorescence staining was used to detect the differentiation of NSCs into neurons. The total number of DAPI-stained nuclei was counted. The phosphorylation of Akt was detected by p-Akt immunofluorescence staining. The expressions of Akt and p-Akt were detected by Western blotting Quantity, and compare each group. Results The ratio of NSCs differentiated into neurons in IGF-1 group was significantly higher than that in control group, LY group and IGF-1 + LY group (all P <0.05). The results of immunofluorescence staining showed that the level of Akt phosphorylation in IGF-1 group was significantly higher In the control group, LY group and IGF-1 + LY group (all P <0.05), Western blotting showed that IGF-1 promoted Akt phosphorylation and LY294002 inhibited Akt phosphorylation. Conclusion IGF-1 can induce the phosphorylation of Akt and activate the PI3K / Akt signaling pathway to promote the differentiation of NSCs into neurons.