基于外泌体miR-337介导TGF-β信号通路探讨祛湿通脉法抑制类风湿关节炎软骨破坏的机制

As known, Rheumatoid arthritis (RA) is an intractable disease to current western and traditional Chinese medicine both. The study team has been insisting on integrating theory of TCM with clinical practice in research. The team proposed that invasion of wind-dampness and blood stasis lead to damage of tendons and bones is the pathogenesis of Rheumatoid arthritis and carried out a series researches on Duanteng Yimu decoction. Based on the researches, the team put forward “ clearing damp and promoting circulation”, which is a treatment method of eliminating wind-dampness, promoting circulation and strengthening tendons and bones, and refined the prescription. Besides, the researches show that the method can significantly improve the clinical symptoms and cartilage destruction of RA. And it is found that the levels of miR-337 decline in the serum、serum exosomes, synovial membrane of RA patients and rise after treatment of the method.In this project, it will explore the key signaling pathways of exosomes miR-337 through studying the differential expression and effect on cartilage destruction of exosomes miR-337 in RA with human RA synoviocytes, chondrocytes and CIA mouse model. Then, it will try to verify that exosomes miR-337 mediated TGF- β signaling to control the RA cartilage destruction is the pharmacological mechanism of clearing damp and promoting circulation. At last, this project may provide scientific evidences for application of the method and development of RA innovative drugs.

类风湿关节炎（RA）是当今中西医学诊疗的疑难疾病。本课题组长期坚持经典理论结合临床实践，提出RA的病因病机为“风湿瘀阻，筋伤骨损”，开展了断藤益母汤系列研究，进一步提出“祛湿通脉法”（祛风湿、通血脉、强筋骨），并优化组方。初步研究表明：祛湿通脉法对改善RA临床症状和软骨破坏有显著疗效；并发现RA患者血清、血清外泌体及滑膜组织中的miR-337水平下降，经祛湿通脉法治疗后其水平上升。本项目拟采用人RA滑膜细胞模型、人软骨细胞模型及CIA小鼠模型，通过观察外泌体miR-337在RA中的差异表达及对软骨破坏的影响，探讨外泌体miR-337抑制RA软骨破坏的关键信号传导通路，以阐明祛湿通脉法通过外泌体miR-337介导TGF-β抑制RA软骨破坏的机制。本项目将为祛湿通脉法临床应用提供科学依据，为RA创新药物研究奠定基础。

类风湿关节炎（RA）是临床最常见的炎性关节炎之一，具有较高的致残率。RA发病后不久就会出现不可逆的关节软骨和骨丢失，软骨侵蚀导致的关节功能受损是致残的主要原因。断藤益母汤治疗RA疗效确切，能减轻滑膜炎症、延缓软骨破坏、保护关节功能，但其保护软骨的作用机制尚不明确。. 课题组前期应用基因芯片及网络药理学方法，发现本方治疗后小鼠与软骨保护相关的miR-337明显上调，我们猜测miR-337可能是断藤益母汤抑制RA软骨破坏的关键因素。软骨没有血管，断藤益母汤通过何种方式作用于软骨细胞，值得深究。已知软骨营养大部分来源于滑液与滑膜，我们猜测断藤益母汤可能先作用于滑膜细胞，再由滑膜细胞递送给软骨细胞起到保护软骨的作用。. 外泌体是当下细胞间物质传递和通讯的热门话题，我们假设断藤益母汤可以通过调节滑膜细胞外泌体miR-337的表达来防治RA软骨破坏。动物实验验证了断藤益母汤对CIA大鼠的软骨保护作用；共聚焦显微镜下观察软骨细胞摄取滑膜细胞上清外泌体，验证了滑膜细胞与软骨细胞之间存在基于外泌体的通讯或物质运输；从体内、体外两方面，验证断藤益母汤诱导的滑膜细胞外泌体具有减轻软骨破坏的功能；通过转染miR-337mimics和inhibitors，验证miR-337是滑膜细胞外泌体携带上软骨细胞发挥软骨保护作用的关键成分。达成了验证外泌体源miR-337在类风湿关节炎软骨分化中的作用及断藤益母汤的干预效果的研究目的。. 研究明确祛湿通脉法通过外泌体miR-337介导TGF-β抑制类风湿关节炎软骨破坏的作用机制，为祛湿通脉法临床应用提供科学依据，为RA创新药物研究奠定基础。