类风湿关节炎滑膜新生血管的Netrin-1体系调控机制及清痹方的干预作用

Netrin-1 is known as a kind of proteins involved in axon guidance, however, recent studies found the Netrin-1 regulating system, constituting Netrin-1 and its receptor family, has a regulatory role in new synovium vascularization of Rheumatoid Arthritis (RA) fibroblast-like synoviocytes as well; the Netrin-1 regulating system plays an important role receptor-dependently in the "death control" molecular mechanism of the process of blood vessel growth and development, and bi-regulation of angiogenesis, while the new synovium vascularization is precisely thought to be a key pathological mechanism of RA. Thus, it can be speculated that the onset and development of RA has an important association with Netrin-1 regulating system. However, no study on the Netrin-1 system role in the regulation of RA synovial neovascularization has been reported by now. Therefore, this study intends to a set CIA animal model and transfected cell researches carrier of Netrin-1 in RA synovial neovascularization process and its receptor, the expression levels of the relevant factors, and the relationship between Netrin-1 regulating system in the RA pathogenesis, in vitro and vivo. At the same time, molecular pharmacology research methods to explore the intervention effect of Qingbi Decoction, following the purpose of TCM'S blood detoxification and collateral dredging, aimed at finding a new target of Chinese medicine treatment of RA and for further research.

Netrin-1是经典的神经突触导向因子，然而新近研究发现Netrin-1及其受体家族构成的体系对类风湿关节炎（RA）成纤维样滑膜细胞的迁移也同样具有调控作用；并且在血管生长发育过程中受体依赖性的参与调控血管新生；而滑膜新生血管异常增生正是类风湿关节炎的关键病理机制，同时已知Netrin-1在RA患者滑膜高表达，故可推测RA的发生发展与Netrin-1调控体系有重要关联。但目前尚无研究报告该体系对RA关节滑膜新生血管形成的调控作用。清痹方治疗RA有确切疗效，基础研究表明其能有效干预滑膜病理进程。因此本研究拟以CIA动物模型、基因干扰细胞为研究载体，研究RA滑膜新生血管增生过程中Netrin-1及其各个受体、相关因子的表达水平和相互关系，进而从体内、体外两条途径阐明该调控体系在RA病理机制中的作用；同时，以分子药理学研究方法探讨解毒通络中药清痹方对该机制的干预作用，旨在寻找中药新靶点。

滑膜新生血管异常增生是类风湿关节炎（RA）的关键病理机制，神经突触导向因子Netrin-1及其受体家族构成的体系对RA成纤维样滑膜细胞的迁移具有调控作用，并且在血管生长发育过程中受体依赖性的参与调控血管新生。本课题以CIA动物模型为研究载体，研究RA滑膜新生血管增生过程中Netrin-1及其各个受体、相关因子的表达水平。实验结果显示，造模后CIA小鼠关节炎症状评分、关节局部肿胀度、X线评分等症状表现，以及关节滑膜炎、血管翳增生等病理学评分明显上升，给予中、高剂量QBD或MTX可抑制关节炎症状进展，并可有效控制关节滑膜炎、血管翳增生，以解毒通络法为核心治法的中药复方QBD各剂量组显示出一定的量效关系，高剂量QBD对该模型的治疗作用近似或优于MTX。可见以解毒通络为主法的清痹方可通过遏制类风湿关节炎netrin-1/受体相关滑膜血管新生、炎症反应等病理变化等机制，对类风湿关节炎具有明确的干预作用。项目成果为临床应用中药治疗类风湿关节炎提供了依据，并为进一步以Netrin-1调控网络相关因子为靶点开发中药新药提供了基础支持。