人骨髓间充质干细胞神经元样细胞分化中自噬相关基因Beclin-1的表达  

Autophagy-related gene Beclin-1 expression in neuron-like differentiation of human bone marrow mesenchymal stem cells

作　　者：杨翼[1] 丁文静[1] 董万利[1] 

机构地区：[1]苏州大学附属第一医院,江苏省苏州市215006

出　　处：《中国组织工程研究》2014年第6期841-846,共6页Chinese Journal of Tissue Engineering Research

基　　金：江苏省自然科学基金项目资助(SBK201123617)~~

摘　　要：背景:骨髓间充质干细胞具有自我更新和多向分化的潜能,但是经过长期体外培养后,骨髓间充质干细胞增殖分化能力逐渐丧失,其机制仍不明确。目的:观察人骨髓间充质干细胞神经元样细胞分化过程中自噬相关基因表达的变化。方法:利用表皮生长因子诱导人骨髓间充质干细胞分化为神经元样细胞,观察其形态变化及生长情况。免疫细胞化学法检测神经元特异性烯醇化酶、胶质纤维酸性蛋白表达,采用Western blot检测诱导前后人骨髓间充质干细胞自噬相关基因Beclin-1表达的变化。结果与结论:经诱导后,人骨髓间充质干细胞呈典型神经元样细胞分化,神经元特异性烯醇化酶阳性率为78.7%,胶质纤维酸性蛋白阳性率为8.1%。诱导0.5 h后处理组细胞中Beclin-1的表达水平有所增加,但1 h后开始逐渐降低。说明体外可以诱导人骨髓间充质干细胞分化为神经元样细胞,自噬相关基因表达活性在诱导分化早期呈高表达,分化过程中逐渐降低。BACKGROUND：Bone marrow mesenchymal stem cells have potential to self-renewal and multi-lineage differentiation. But after a long period of culture in vitro, the proliferation and differentiation capacities of bone marrow mesenchymal stem cells gradual y loss, the mechanism underlying which is not clear now. OBJECTIVE：To observe the expression of autophagy-related gene Beclin-1 in differentiation from human bone marrow mesenchymal stem cells into neuron-like cells in vitro. METHODS：The changes of morphological characteristics of neuron-like cells differentiated from human bone marrow mesenchymal stem cells induced by epidermal growth factor were observed. The expression of neuron-specific enolase and glial fibril ary acidic protein in treated and untreated human bone marrow mesenchymal stem cells were detected using immunocytochemistry. The Beclin-1 protein expressions were detected by western blot before and after induction.RESULTS AND CONCLUSION：After being induced, human bone marrow mesenchymal stem cells presented classical neuron-like morphology;the expressions of neuron-specific enolase and glial fibril ary acidic protein were 78.7%and 8.1%, respectively. The expression of Beclin-1 protein was changed correspondingly during the induction, which increased after 30 minutes of induction and decreased gradual y after 1 hour of induction. Human bone marrow mesenchymal stem cells could be induced into neuron-like cells in vitro by epidermal growth factor. Autophagy-related gene was highly expressed in the induction of early differentiation and the expression gradual y reduced until it remained at a low level during the differentiation.

关 键 词：干细胞 骨髓干细胞 人骨髓间充质干细胞 神经元样细胞分化 体外实验 表皮生长因子 自噬 BECLIN-1 江苏省自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]