Clinical Efficacy Study And Mechanism Research On Rheumatoid Arthritis Treated By WenYangSanHanChuShi Method

Objective To study the clinical effects, efficacy mechanism and security of WenYangSanH-anChuShi method(Juan Bi particle) on the treatment of rheumatoid arthritis(RA), to provide theoretical basises for WenYangSanHanChuShi method in RA and enrich theories of treating RA with traditional Chinese medicine.Methods①Clinical studies cited internationally recognized DAS28, CDAI, SDAI disease activity index to evaluating the curative effect.There are totally102cases of RA patients with cold-dampness syndrome in this study, which are randomly divided into treatment group of68cases and control group of34cases. Treatment group use Juanbi particle(45g/d),and the control group use Zhengqing Fengtongning Retard tablets(120mg/d). Each group take a8weeks treatment, after that,observing the clinical efficacy and safety.②The models of carrageenan-induced edema of rat plantar and formation of cotton ball granulom in rats were adopted to investigate anti-inflammatory of WenYangSanHanChuShi method. Rats were divided into five groups, including the model group, the positive control group(DEX) and Juanbi particle groups(low, medium and high dose group). After drug intervention, effects on acute and chronic inflammation was evaluated.③Type ⅡII bovine collagen(CII)-induced arthritis (CIA) in rats were established for investigating the anti-synovitis of WenYangSanHanChuShi method. Rats were divided into six groups, including the blank control group, model group, the positive control group(MTX) and Juanbi particle groups(low, medium and high dose group).After drug intervention, arthritis disease degree was evaluated by arthritis index (AI) score method. The histological damage of synovial tissues was assessed by histological examination (H&E).Serum level of IL-1β was detected by enzyme-linked immunosorbent assay(ELISA).④Type Ⅱ bovine collagen(CII) induced arthritis (CIA) in mice were established for evaluating the inhibitory effect and mechanism of WenYangSanHanChuShi method. Mice were divided into three groups, including the model group, the positive control group(MTX) and Juanbi particle groups by the clinical score method. After drug intervention, arthritis disease degree was evaluated by the clinical score. The histological damage of arthrosis was assessed by histological examination (H&E) and X-ray mammography radiography. Lymphocyte proliferative ability in CIA mouse was detected by3H-thymidine incorporation assay. The inflammatory cytokine production level of IL-6,IL-17A and TNF-a in collagen(CII)-induced splenic lymphocytes was detected by Cytometric Bead Array (CBA) and IFN-y, IL-10was detected by ELISA. Surface marks of spleen lymphocyte(CD4, CD8, B220ratio) and the level of IFN-y, IL-17A in spleen lymphocytes was detected by fluorescence activated cell sorter(FACs).Results①Clinical observation results show that the syndrome curative effect of treatment group was better than control group. About improvement of TCM syndrome integral, CRP, ESR, DAS28, CDAI, and SDAI, it was better than the control group (P<0.01, P<0.05),and did not appear obvious side effects and adverse reactions.②WenYangSanHanChuShi method has significant inhibitory effect on swelling of paw and the formation of cotton ball granuloma(P<0.05).③Improving arthritis AI score(P<0.05), joint synovial inflammation (P<0.01), synovial tissue inflammatory cells infiltration and tissue hyperplasia and reducing levels of serum IL-1β(P<0.01) in CIA rats.④Improving clinical scores(P<0.01), reducing arthritis cell infiltration and bone injury, inhibiting immune response collagen induced(P<0.01), reducing the level of IL-17A in intrace-extracellular spleen lymphocytes collagen induced, raising IL-10in collagen induced splenic lymphocytes(P<0.01) and Promoting Treg cell expression in some degree. But there is no obvious influence on the level of IFN-y, TNF-a, IL-6and lymphocyte subgroups(CD4, CD8, B220ratio) in CIA mice.Conclusion①WenYangSanHanChuShi method is an effective measure in treatment of RA.②WenYangSanHanChuShi method has an obvious anti-inflammatory effect.③It has a therapeutic effect on CIA rats and CIA mice.④The upregulation of expression of IL-10and the downturn of expression of IL-17A in collagen(CII)-induced splenic lymphocytes may be one of the mechanisms in treatment of RA with WenYangSanHanChuShi method.