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目的:探讨细胞外信号调节激酶1/2(ERK1/2)信号通路在胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)诱导新生小鼠神经干细胞(neural stem cells,NSCs)增殖分化中的作用。方法:取新生C57BL/6J小鼠海马体外分离、培养NSCs,分别加入终浓度100 ng/ml的IGF-1和20μmol/L的抑制剂U0126。CCK-8试剂盒检测IGF-1对NSCs增殖的影响;免疫荧光细胞染色法检测IGF-1对细胞分化的影响;Western blot检测ERK1/2和p-ERK1/2蛋白的表达,并对各组进行比较。结果:CCK-8测定细胞增殖,第3 d U0126组相对OD值显著低于IGF-1组,第7 d时IGF-1组OD值则显著高于对照组和U0126组(P<0.05)。倒置荧光显微镜下可观察到IGF-1组βⅢTubulin和GFAP阳性分化率均明显高于对照组和U0126组(P<0.05)。Western blot结果显示:IGF-1组蛋白表达量显著高于对照组和U0126组(P<0.05),IGF-1促进ERK磷酸化,U0126则抑制ERK的活化。结论:IGF-1可显著诱导NSCs的增殖和分化,ERK1/2信号通路可能具有重要作用,同时IGF-1可能参与ERK1/2的活化。
OBJECTIVE: To investigate the effect of extracellular signal-regulated kinase 1/2 (ERK1 / 2) signaling pathway on neural stem cells (NSCs) induced by insulin-like growth factor-1 (IGF- ) Proliferation and differentiation in the role. METHODS: NSCs were isolated from hippocampus of neonatal C57BL / 6J mice. IGF-1 (100 ng / ml) and U0126 (20 μmol / L) were added respectively. The effect of IGF-1 on the proliferation of NSCs was detected by CCK-8 kit. The effect of IGF-1 on cell differentiation was detected by immunofluorescence staining. The expressions of ERK1 / 2 and p-ERK1 / 2 were detected by Western blot. Compare. Results: Compared with IGF-1 group, the relative OD value of U0126 group on day 3 was significantly higher than that of IGF-1 group on the 7th day (P <0.05). Under inverted fluorescence microscope, the positive rate of βⅢTubulin and GFAP in IGF-1 group was significantly higher than that in control group and U0126 group (P <0.05). Western blot results showed that IGF-1 protein expression was significantly higher than that of the control group and U0126 group (P <0.05), IGF-1 promoted ERK phosphorylation, and U0126 inhibited ERK activation. Conclusion: IGF-1 can significantly induce the proliferation and differentiation of NSCs. ERK1 / 2 signaling pathway may play an important role, and IGF-1 may be involved in the activation of ERK1 / 2.