Kartogenin预处理促进TGF-β3诱导骨髓间充质干细胞成软骨分化及阻抑骨化的机制研究

项目名称： Kartogenin预处理促进TGF-β3诱导骨髓间充质干细胞成软骨分化及阻抑骨化的机制研究

项目编号： No.81501604

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： 何晓敏

作者单位： 上海交通大学

项目金额： 18万元

中文摘要： 如何有效促进骨髓间充质干细胞（BMSCs）成软骨分化并维持软骨细胞表型以避免骨化是目前软骨组织工程研究的热点和难点。我们前期研究发现BMSCs经小分子化合物Kartogenin（KGN）体外预处理一定时间，再利用TGF-β3常规诱导可以促进成软骨并抑制骨化，但具体机制不明。KGN与RUNX1密切相关，且已有证据表明RUNX1/RUNX2失衡是引起软骨骨化的关键因素。本项目拟在前期研究基础上，探讨RUNX1/RUNX2信号通路在KGN预处理促进BMSCs成软骨及阻抑骨化中的作用，并利用蛋白质芯片技术筛选相关信号通路靶标，发现与调控RUNX1/RUNX2信号通路可能相关的蛋白质分子，再运用基因过表达、沉默等技术验证所筛选的信号分子对RUNX通路的调节作用，以阐明KGN预处理促进TGF-β3诱导BMSCs形成更为稳定的软骨组织并阻抑软骨内骨化的作用机制，为组织工程软骨走向临床应用提供实验依据。

中文关键词： 骨髓间充质干细胞；kartogenin；软骨组织工程；信号通路；骨化

英文摘要： How to effectively promote the bone marrow mesenchymal stem cells (BMSCs) differentiation into chondrocytes and maintain the chondrocytes phenotype in order to avoid ossification are the hotspot and difficult problem in cartilage tissue engineering. We have previously found that BMSCs could be promoted to chondrogenesis and inhibitted for ossification when BMSCs were pretreated by a new small molecule named kartogenin (KGN) in vitro, and then induced by traditional inducing factor of transforming growth factor β3 (TGF-β3). However, the pathways KGN promoted BMSCs chondrogenesis and inhibitted ossification were still unclear. The target of KGN for signaling pathway was runt related transcription factor 1(RUNX1). Besides, the evidences have shown that the imbalances of RUNX1/RUNX2 were key factors to cause cartilage ossification. Therefore, this project will further clarify the relationship of KGN and RUNX1/RUNX2 in based of preliminary experiment. Furthermore, the related signaling pathway will be screened by protein chip, in order to find related protein molecules which may regulate RUNX1/RUNX2 signaling pathway. Finally, these screened protein molecules wil be verified by in vitro cell biology experiment to clarify the molecular mechanism of KGN in chondrogenic differentiation and ossification inhibition of BMSCs induced by TGF-β3. This project will provide the experimental basis for the clinical application of cartilage tissue engineering.

英文关键词： Bone marrow mesenchymal stem cells;Kartogenin;Cartilage tissue engineering;signaling pathway;Ossification