基于TLR4/NF-κB通路研究“滑膜-软骨”轴对膝骨关节炎的发病作用及利湿化瘀方干预机制

Knee osteoarthritis (OA) as a degenerative joint disease, is caused by many factors accompanied with articular cartilage damage and demyelination, subchondral sclerosis and synovial hyperplasia, etc. New knowledge of pathological "synovium-cartilage" axis derives from abnormal immune-mediated synovial changes to cartilage degeneration of OA by a link of metabolic inflammatory cytokines. This utilizes us to understand the pathogenesis of OA and to develop therapeutic strategies with a new perspective. Previous clinical studies have confirmed the acceptable practice of Lishi Huayu decoction in the treatment of OA. Combined with activation of TLR4/NF-κB signaling pathway, its inhibition is an important way for OA synovial hyperplasia, Lishi Huayu decoction has one of the key aspects for OA treatment. The rat OA model will be used with histopathology, immunohistochemistry and methods in molecular biology to observe: (1) the dynamic pathological changes of "synovium-cartilage" axis and their relevancies; (2) the intervention of Lishi Huayu decoction for "synovium-cartilage" axis. Not only this study can further reveal mechanisms of therapy theory of "Lishi Huayu" to treat OA, but also may provide the new evidence for "synovial membrane" as a therapeutic target.

膝骨关节炎（OA）是由多种因素引起关节软骨破损、脱失，软骨下骨硬化及滑膜增生等而导致的关节退行性病。以免疫异常介导滑膜改变为源头，代谢性炎症因子为纽带，软骨退变为病理结果的OA“滑膜-软骨”轴新认识，为我们理解OA发病机理及制定治疗策略提供了新的视角。既往临床实践和研究证实利湿化瘀方有理想的治疗OA作用，结合TLR4/NF-κB信号通路活化，提出滑膜是OA治疗的重要靶位，也是利湿化瘀方治疗OA的关键环节之一。拟采用大鼠OA模型，通过组织病理学、免疫组化及分子生物学方法观察：（1）“滑膜-软骨”轴病理环节的动态变化及关联性；（2）利湿化瘀方对上述病理环节的干预作用。该课题不仅可以进一步揭示“利湿化瘀”治疗OA的作用机理，而且有可能为“以滑膜为靶位”的治疗策略提供新的证据。

滑膜是膝骨关节炎（OA）发病的重要因素之一，滑膜炎症和软骨退变相互影响。先天免疫通过TLR4/NF-κB通路过度活化，使炎症因子等指标的表达升高引起滑膜炎症，并介导关节软骨破坏，从而诱发OA的发生和发展。抑制TLR4/NF-κB 信号通路为我们认识OA 发病机理及制定治疗策略提供了新的视角。因此我们采用ACLT法建立大鼠膝OA模型，8周龄雄性SD大鼠随机分成正常组、模型组、塞来昔布胶囊组、利湿化瘀方组（高、中、低剂量组）。除正常组外，其余各组采用Hulth法建立膝OA动物模型。造模后，各实验组给予相应的药物灌胃，正常组、模型组给予等量生理盐水。术后4天、3周、6周随机选取各组3只处死，比较各组关节软骨和滑膜病理变化，基因及蛋白表达改变，以及利湿化瘀方的干预机制。.研究发现模型组的病理改变明显重于正常组。膝OA关节滑膜中CD14，TLR-4，IL-1β和TNF-α，ADAMTS-4，MMP-13基因、NF-κB p65 蛋白呈高表达变化。血清中PIIINP 及 HA 浓度的高于正常组。随造模时间的延长，两组间 CTX-II、COMP、PIIANP、IL-1β、TNF-α浓度均无显著变化。同时，我们发现随着NF-κB信号通路激活，软骨损伤的越重。.利湿化瘀方低、中、高剂量组的滑膜炎症和软骨损失明显小于同期模型组，但大于正常组。利湿化瘀方高剂量组的软骨退变及滑膜炎症较轻，并优于塞来昔布组。利湿化瘀方显著抑制外周血中炎症因子PIIINP及PIIANP，而对HA、CTX-II、TNF-α及IL-1β的浓度变化无明显影响。利湿化瘀方能明显抑制TLR4/NF-κB 信号通路的激活。.本课题已验证了退变性之OA，以先天免疫为“启动子”，滑膜改变为源头，滑膜是OA治疗的重要靶位，也是利湿化瘀方治疗OA的关键环节之一。在OA病理机制的探讨以及中医药防治OA方面都具有较好的科学价值，并为OA防治提供新证据。