补肾活血汤提取物对MSCs迁移过程中Wnt信号通路的分子机理研究

项目名称： 补肾活血汤提取物对MSCs迁移过程中Wnt信号通路的分子机理研究

项目编号： No.81473699

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 罗毅文

作者单位： 广州中医药大学

项目金额： 72万元

中文摘要： 本项目拟采用Transwell体外迁移实验观察补肾活血汤石油醚、乙酸乙酯、无水乙醇、水溶性4个提取部位对MSCs迁移能力的影响，选取促进MSCs迁移的特定活性部位；根据选取的特定活性部位继续观察补肾活血汤提取物对MSCs相关趋化因子及其受体的表达；采用RT-PCR、Western Blot等方法，研究补肾活血汤在MSCs迁移过程中Wnt信号通路作用的影响；运用荧光素酶报告基因技术，观察Wnt5a基因过表达对MSCs迁移能力的影响；进一步运用siRNA技术，干扰Wnt5a在MSCs的表达，观察对MSCs迁移能力的影响。期望通过本项目的研究，进一步丰富中医肾主骨、髓生骨理论，从而为中医药治疗骨缺损、骨不连及骨折长期制动后出现的废用性骨质疏松等难治性骨病提供新思路和理论依据。同时，深入发现补肾活血汤干预骨折愈合的药理学作用，从而为今后中医临床治疗骨折药物的评价与筛选提供药理学依据。

中文关键词： 补肾活血汤；骨髓间充质干细胞；迁移；Wnt信号通道；分子机理

英文摘要： The Transwell assay will be used to identify the active components of the BuShenHuoXueTang(BSHXT) involved in promoting migration of MSCs. The BSHXT is extracted with petroleum ether,ethyl acetate,absolute alcohol and aqueous respectively.And this active component will be used to research the expression of MSCs related chemotactic factor and its receptor.The influence of Wnt signaling pathway affected by the active component will be analyzed with the RT-PCR,Western Blot etc. Subsequently,the migration capacity of MSCs treated either with Wnt5a over-expression or siRNA against Wnt5a will be analyzed with the luciferase reporter assay and Transwell assay in the further study. We hope this study could further enrich the traditional Chinese theory of kidney governs bone and marrow giving birth to bone,and offer traditional Chinese medicine the new ideas and theoretical basis in the treatment of refractory bone diseases like bone defect,bone ununion and disuse osteoporosis after long terms immobilization. Meanwhile,the pharmacological activities that BSHXT promotes fracture healing could be found in-depth,and pharmacological evidence in evaluating and screening of chinese medicine in the treatment of bone fractures could be provided.

英文关键词： BuShenHuoXueTang extracts;MSCs;migration;Wnt signaling pathway;molecular mechanism