Notch信号系统对骨髓间充质干细胞向GABA能神经元分化的调节作用及机制研究

项目名称： Notch信号系统对骨髓间充质干细胞向GABA能神经元分化的调节作用及机制研究

项目编号： No.31301216

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 龙乾发

作者单位： 西安交通大学

项目金额： 21万元

中文摘要： 研究证实骨髓间充质干细胞(BMSCs)可向GABA能神经元分化，且我们前期研究发现Notch信号通路靶基因Hes1沉默可促进BMSCs的这一分化潜能，但是具体机制不清。本课题拟首先通过条件诱导BMSCs向GABA能神经元分化，建立标准分化方案；然后依赖Notch信号通路阻断或效应器沉默，观察其对GABA能神经元生成的影响，确定Notch信号系统是否参与调节BMSCs向GABA能神经元分化；继而通过检测Hes1及其密切相关的上/下游基因（RBPJ/Mash1）在BMSCs分化前后的表达变化，明确其参与调节方式；而后采用RNA干扰或过表达上述基因，体内外观察其对GABA能神经元生成的影响，分析Hes1及其上/下游基因在该分化中的调节作用，最终阐明Notch信号系统对BMSCs向GABA能神经元分化的调节机制。阐明该机制将拓展BMSCs向GABA能神经元分化的潜能，为干细胞治疗癫痫开创新路径。

中文关键词： 骨髓间充质干细胞；伽马氨基丁酸能神经元；Notch信号通路；神经母细胞特异性转移因子1；癫痫

英文摘要： Evidence to date has shown that bone marrow mesenchymal stem cells (BMSCs) are capable of differentiating into gamma aminobutyric acid (GABA)ergic neurons,and our previous study indicated that Hes1 gene (which is the target of Notch signaling pathway) silencing can promote the GABAergic differentiation of BMSCs. But, the mechanism of GABAergic differentiation of BMSCs is not clear.This study was intended to establish the standard method which induced BMSCs to differentiate into GABAergic neurons firstly.Then to confirm whether Notch signals participated in the GABAergic differentiation of BMSCs or not,we examined the influence on GABAergic differentiation by inhibiting Notch signaling pathway or silencing its effector. Furthermore, we evaluated the regulatory role of Hes1 gene and its closely related upstream (RBPJ) and downstream (Mash1) gene in the Notch signals by analyzing the expression change of these genes in the process of GABAergic differentiation of BMSCs. Additionally, to further investigate the regulatory role of RBPJ, Hes1 and Mash1 gene in the GABAergic differentiation of BMSCs, we examined the influence on GABAergic differentiation with RNAi or overexpression of the RBPJ, Hes1 and Mash1 gene in BMSCs in vitro and in vivo. Upon the results of these essays, we would clarify the regulatory mechanism

英文关键词： bone marrow mesenchymal stem cells；GABAergic neuron；Notch signaling pathway；mammalian achaete-scute homologue 1；epilepsy