ERK1/2信号通路在IGF-1诱导新生小鼠海马神经干细胞增殖分化中的作用  

The role of ERK1/2 signal pathway in IGF-1 inducing proliferation and differentiation of neural stem cells in hippocampus of neonatal mice

作　　者：朱裕华[1] 胡安康[1] 陈仁金[1] 彭长凌[1] 吴连连[2] 袁红花[2] 朱孝荣[1] 

机构地区：[1]徐州医学院实验动物中心,徐州221002 [2]徐州医学院神经生物学教研室,徐州221002

出　　处：《神经解剖学杂志》2013年第6期609-614,共6页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(31172171);江苏省自然科学基金(BK2011209);江苏省青年基金(BK2012138)

摘　　要：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 role of extracellular signal-regulated kinase 1/2 （ERK1/2） signal pathway in insulin-like growth factor-1 （IGF-1） inducing proliferation and differentiation of neural stem cells （NSCs） in neonatal mice. Methods： Neonatal hippocampus of C57BL/6J mice were isolated to culture NSCs in vitro, and then the NSCs were added to a final concentration of 100 ng/ml IGF-1 and 20 μmol/L inhibitor U0126. Detecting the proliferation of NSCs added IGF-1 with CCK-8 kit; detecting the differentiation when IGF-1 acting on cells by immunofluorescence histo- chemistry; detecting the expression of ERK1/2 and p-ERK1/2 protein with Western blot, and then compared each group. Results： CCK-8 determination of cell proliferation, the relative OD value in U0126 group was lower than that in IGF-1 group in the third day, OD value was significantly higher in IGF-1 group than that of control group and U0126 group in the seventh day （ P 〈 O. 05 ）. Under inverted fluorescence microscope, ~ III Tubulin and GFAP positive differentiation rate in IGF-1 group were significantly higher than those in control group and U0126 group （ P 〈 O. 05 ）. Western blot results showed that the expression protein content in IGF-1 group was significantly higher than that in control group and U0126group （P 〈0.05）. IGF-1 promoted the phosphorylation of ERK and U0126 inhibited ERK activation. Conclusion： IGF- 1 can significantly induce NSCs proliferation and differentiation, which may play an important role in ERK1/2 signaling pathway, and IGF-1 may be involved in ERK1/2 activation.

关 键 词：ERK1 2信号通路 胰岛素样生长因子-1 神经干细胞 增殖和分化 小鼠 

分 类 号：R338[医药卫生—人体生理学]