miR34a与PI3KⅢ通路crosstalk调控成骨在补肾法治疗激素性骨质疏松症中的机制

项目名称： miR34a与PI3KⅢ通路crosstalk调控成骨在补肾法治疗激素性骨质疏松症中的机制

项目编号： No.81503591

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 江晓兵

作者单位： 广州中医药大学

项目金额： 18万元

中文摘要： 我们前期研究表明成骨细胞（OB）骨形成功能低下在激素性骨质疏松症（GIOP）发病中起关键作用。miR34a是一类非编码单链小RNA分子，直接调控骨形成。PI3KⅢ通路启动自噬可调节OB的活性和分化。且miR34a与PI3KⅢ通路crosstalk已被证实，但在GIOP中尚无报道。补肾法可促进成骨有效治疗GIOP，但其深层机制仍待明确。因此，我们提出“补肾中药通过miR34a与PI3KⅢ通路crosstalk调控成骨治疗GIOP”的假说。本研究拟用透射电镜、qPCR、蛋白印迹、免疫共沉淀等技术从不同层面进行验证：①体内探讨miR34a与PI3KⅢ通路在补肾法促GIOP骨形成中的作用；②体外GIOP的OB实验中过表达或沉默miR34a和激活或阻断PI3KⅢ通路探讨补肾法调控miR34a与PI3KⅢ通路crosstalk促骨形成的机制,为丰富GIOP发病机理及“肾主骨”理论的科学内涵提供依据。

中文关键词： 补肾法；；miR34a；PI3KⅢ通路；糖皮质激素性骨质疏松症；成骨细胞

英文摘要： Our pilot studies showed that bone formation deficiency of Osteoblast(OB) played a key role in the development of Glucocorticoid-induced osteoporosis(GIOP). MiR34a is a short non-protein coding RNA that regulates directly bone formation. PI3KⅢ pathway activating autophagy is a new player in regulating the activity and differentiation of OB. Moreover, the crosstalk between miR34a and PI3KⅢ pathway has been confirmed in the other studies, but its role in GIOP is still unclear. Tonifying kidney could promote bone formation for treating GIOP effectively, however, the underlying mechanism of action still need to be investigated. Therefore, we hypothesize that the Chinese herbs of tonifying kidney regulate bone formation to treat GIOP via the crosstalk between miR34a and PI3KⅢ pathway.To verify the hypothesis, the techniques, including transmission electron microscope, qPCR, Western-blot, Immunoprecipitation, etc, will be employed in two main experiments from different levels, one is in vivo GIOP rats model study in which the role of miR34a and PI3KⅢ pathway in the process of tonifying kidney promotes the bone formation will be clarified, and one is in vitro osteoblast cell experiments in which overexpression or silence of miR34a and activation or inhibition of PI3KⅢ pathway will be used to investigate that the mechanism of tonifying kidney regulate the crosstalk between miR34a and PI3KⅢ pathway to activate bone formation. This study will provide the theoretical basis to enrich the pathogenesis of GIOP and the scientific connotation of “kidney controlling bone” theory.

英文关键词： tonifying kidney; miR34a;PI3KⅢ pathway;glucocorticoid-induced osteoporosis;osteoblast