基于骨免疫探讨补肾通络法对类风湿关节炎骨破坏及Treg/Th17的影响及机制研究

Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by joint destruction and joint dysfunction. Clinical treatment takes control of the disease and inhibition of bone destruction as the goal. Kidney deficiency and blocks in main and collateral channels are important factors in rheumatoid arthritis bone destruction. Our previous studies have shown that Bushen Tongluo therapy can significantly reduce the level of MMP-3, TNF-a in RA rats; clinical studies have shown that Bushen Tongluo therapy can delay RA bone destruction and is effective in clinical treatment. This project intends to take bone metabolism and immune balance as the breakthrough point. Through the technology of Western blot, RT-PCR, pathology and imaging evaluation, this project will establish osteoclast differentiation model in vitro, observe the effect of Bushen Tongluo therapy on osteoclast differentiation, bone resorption function and Treg / Th17, explore the role and molecular mechanism of Bushen Tongluo therapy in RA bone destruction and bone immunology from such aspects as gene, mRNA and protein level, comprehensively expound the modern biological characteristics and internal laws of " Bushen Tongluo therapy ", and enrich the scientific theory of "treating diseases from the kidney and adjusting the whole" of RA TCM to guide the clinical practice more effectively.

类风湿关节炎（RA）是一种以关节破坏和关节功能障碍为特点的全身性自身免疫性疾病。临床以控制病情、抑制骨破坏为治疗目标。肾虚络阻是类风湿关节炎骨破坏的重要因素，我们前期研究表明补肾通络方药能明显降低RA大鼠MMP-3、TNF-a的水平，临床研究证实补肾通络方药具有延缓RA骨破坏的作用，临床治疗RA骨破坏疗效显著。本课题拟以调节骨代谢及免疫平衡为切入点，通过Western blot、RT-PCR、关节病理及影像学评价的技术，建立体外破骨细胞分化模型,观察补肾通络方对破骨细胞分化、骨吸收功能及Treg/Th17的影响，从基因、mRNA、蛋白水平多层次探讨补肾通络方在RA骨破坏及骨免疫学的作用及分子机制，全面阐释“补肾通络法”理论的现代生物学特征及内在规律，丰富RA中医“从肾论治，整体调节”的科学理论，更有效的指导临床实践。

RA是一种以对称性、多关节的炎症性滑膜炎和骨侵蚀为主要特征的慢性自身性免疫性疾病,肾虚络阻是RA骨破坏的重要病机，补肾通络方前期应用于临床，在延缓RA骨破坏、促进骨修复，防止关节变形，提高患者生存质量等方面取得了独特疗效。本课题拟以调节骨代谢及免疫平衡为切入点，观察各组大鼠一般状态、踝关节活体CT、滑膜病理切片以及采用ELISA法、Western blot法、R-T PCR法测定相关炎性因子和骨破坏相关蛋白的表达，观察补肾通络方对破骨细胞分化、骨吸收功能及Treg/Th17的影响。实验结果表明补肾通络方可能通过调节RANKL-RANK-OPG信号通路以及调节Th17/Treg细胞失衡、抑制促炎因子，从而有效控制CIA大鼠体内炎性程度和骨破坏的发展，对RA有一定的治疗作用，为补肾通络方临床用药提供了理论依据。