Rho相关卷曲螺旋蛋白激酶通路阻断剂Y-27632对大鼠脂肪组织源性干细胞向神经元样细胞分化的诱导作用  

Potentiation of neuroal-like differentiation from rat adipose mesenchymal stem cells by Rho-associated coiled kinase inhibitor Y-27632

作　　者：任宏媛[1] 徐洪[2] 于婉莹[2] 袁媛[2] 孙月[2] 杨方[2] 吴庆文[1] 

机构地区：[1]河北联合大学护理与康复学院,唐山063000 [2]河北联合大学,医学实验研究中心,唐山063000

出　　处：《解剖学杂志》2013年第4期772-774,809,F0003,共5页Chinese Journal of Anatomy

摘　　要：目的：研究Y-27632对大鼠脂肪间充质干细胞向神经元样细胞诱导分化作用.方法：原代培养大鼠脂肪间充质干细胞,将3～6代细胞随机分成4组,无血清对照组、无血清诱导组、血清对照组、血清诱导组.采用倒置显微镜观察细胞形态,免疫细胞化学检测巢蛋白、神经元特异性核心抗原（NeuN）、血清应答因子（SRF）、平滑肌肌动蛋白（α-SMA）、vimentin的表达,免疫印迹检测SRF、vimentin的表达.结果：在无血清情况下,Y-27632能够诱导脂肪组织源性干细胞（ADSCs）出现典型的神经元样细胞形态,免疫细胞化学检测显示神经元特异性蛋白巢蛋白、NeuN的阳性表达；而在10％血清条件下,这种诱导效应被抑制,巢蛋白、NeuN无阳性表达.免疫细胞化学染色显示无血清条件下,Y-27632能够显著抑制α-SMA的表达,同时SRF和vimentin也具有较强的阳性表达.免疫印迹检测显示无论是否存在血清,Y-27632均能够抑制SRF的表达；vimentin在无血清＋Y-27632组表达高于其他各组.结论：Y-27632能够诱导ADSCs向神经元样细胞分化并伴随神经特异标记物的表达.Objective： To investigate the effect of Y-27632 in the neuroal-like differentiation from adipose tissue-derived stem cells（ADSCs）. Methods： ADSCs were cultured from adult rats and the 3-6 passage cells were divided into four groups： serum-flee control, serum-free＋Y-27632, 10% serum control, 10% serum＋Y-27632. Morphological changes of the ADSCs were observed under a microscope. The expressions of the nestin, NeuN, SRF, a-SMA and vimentin were detected by immunohistochemistry, and the expressions of SRF and vimentin were measured by Western blotting. Results： Y-27632 had a pontency to induce neuronal-like differentiation from ADSCs in serum-free condition, with a positive expression of nestin and NeuN and an inhibition of a-SMA, while this inductive effect was suppressed by 10~ serum. The ADSCs treated with Y 27632 also induced a positive expression of SRF and vimentin. However, Western blotting showed that Y-27632 inhibited the expression of SRF and induced the expression of vimentin in the serum-flee condition. Conclusion： Y-27632 can induce ADSCs differentiation into neuron-like cells with the expression of neural specific markers in vitro.

关 键 词：Rho相关卷曲螺旋蛋白激酶通路阻断剂 脂肪间充质干细胞 神经元样分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]