Aβ_(25-35)对体外培养大鼠神经干细胞分化的影响  

The Effect of Aβ_(25-35) on Differentiation of Mice Neural Stem Cells in Vitro

作　　者：蒋珂[1,2] 李博[1] 郭思[1,3] 段萍[1] 韩雪飞[1] 许燕[1] 鄢文海[1] 邢莹[3] 

机构地区：[1]郑州大学基础医学院,郑州450001 [2]商丘市第一人民医院神经内科,河南商丘476100 [3]新乡医学院,河南新乡453003

出　　处：《河南职工医学院学报》2014年第1期1-5,共5页Journal of Henan Medical College For Staff and Workers

摘　　要：目的探讨β淀粉样蛋白(Aβ25-35)对体外培养神经干细胞分化的影响和机制。方法分离培养新生SD大鼠经干细胞,加入10%的胎牛血清使其分化7 d时,一部分细胞用Aβ25-35作用12 h,观测细胞生长状态,运用免疫细胞化学和免疫蛋白印记迹术研究各组Tau蛋白在Ser396,Ser262位点磷酸化水平变化及活化型GSK-3β[pTyr279,216]的表达水平。结果呈球形生长的神经干细胞在加入胎牛血清培养液后,逐渐从球的周边分化出神经细胞,长出神经细胞样的触突,在Aβ25-35作用下,分化细胞的突起粗大且生长速度较快。免疫细胞化学染色和Western-blot结果显示,所有分化细胞都有Tau[pS396]、Tau[pS262]、GSK-3β[pTyr279,216]表达,但经Aβ处理后表达量增高。结论 Aβ25-35可通过上调GSK-3β的表达提高Tau蛋白磷酸化水平,促进神经干细胞向神经细胞分化。Objective To explore the effect and the mechanism of Aβ25-35 on the process of differen- tiation of neural stem cells. Methods Neural stem cells were isolated and cultivated from the neonatal rat hippocampus dentate gyrus and added by 10% FBS. After 7 days, some cells were used to be acted with Aβ25-35 for 12h and their growth states were observed. Immuocytochemistry and Wstern-blot were used to explore and investigate the expression of Tau [ pS396 ], Tau [ pS262 ], GSK-3β [ pTry^279,216] in the two groups. Results After the neural stem cells which grow spherically were added with FBS, neural stem cells were differentiated gradually from neurospheres and neuron cell-like synapses were grown. Un- der the effect of Aβ25-35, the cell projection of differentiated cells becomes thick and grow rapidly. The results of chemical staining on immune cells reveal that all differentiated cells have the expression of Tau [ pS396 ], Tau [ pS262 ], GSK - 3β [ pTry^279,216 ]. However, the expression magnitude increased after processed with Aβ25-35. Conclusion Aβ25-35 could raise Tau protein phosphorylation levels by adjusting up the expression of GSK-313 and promote the process of differentiation from neural stem cells to neural cells.

关 键 词：Β淀粉样蛋白25-35 神经干细胞 TAU蛋白 糖原合成酶3β 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]