Clinical And Mechanism Research Of Qingrehuoxue Therapy Treating Active Rheumatoid Arthritis

Rheumatoid arthritis (RA) is a chronic, systemic inflammatory disorder that may affect many tissues and organs, but principally attacks synovial joints. The pathology of the disease process often leads to the destruction of articular cartilage and bone, finally the affected joints are ankylosis. About1%of the world’s population is afflicted by RA. Onset is most frequent between the ages of40and50, but people of any age can be affected.70%patients with RA have joint erosion.90%patients with RA have joint destruction just2years after the onset of the symptoms.The primary goal of RA treatment is through control inflammation symptoms, prevent joint destruction, keep joint function, maintain normal social activities and maximize the long-term health of the patients, quality of life, and delay/block RA bone destruction is the ultimate goal of treatment. RA disease activity is major factor of bone destroy, the inflammatory process affect not only local bone, can also lead to far away from the parts of the bone loss, hints that bone destruction and RA synovial membrane inflammation has close relationship. Our department according to active period of pathogenesis of RA characteristics combined with years of the clinical experience, emphasis on heat and dampness stasis is the important active RA pathogenic mechanism and pathological key of bone damage, Qingrehuoxue therapy is basic treatment method of active period RA.This project under the guidance of Qingrehuoxue therapy, scientific development in the capital fund Beijing medical medical health technology project and the national natural science funds subject as the basis, used the immunohistochemical, ELISA, cell co-culture technologies, from the theory research, clinical research and the experimental research three aspects to explore effectiveness of Qingrehuoxue decoction treatment RA to reduce desease activity and delay bone destruction, and the internal mechanism was also be discussed.Therapeutic Study1. By sorting out the Traditional Chinese Medicine historical evolution, know RA related problems and symptoms and etiology, the history of the understanding, combed the disease and syndrome with combined therapy of diagnosis and treatment of RA ideas.2. To analyze the "heat, dampness and blood stasis to bi" relevant literature and modern research results, and summarized heat, dampness and blood stasis are the key TCM pathogenesis of active RA, and is the key pathogenesis of bone destruction, put forward that Qingrehuoxue therapy is active RA treatment method.3. Through sorting "bone" the physiology, pathology and treatment in the Traditional Chinese Medicine literature, discusses and RA bone destruction etiology and pathogenesis, the paper summarized the destruction of RA bone clinical and mechanism research progress, and puts forward the clinical and bone is destroyed RA mechanism study of the various ideas, enrich the theory of traditional Chinese medicine treatment RA.Clinical StudyObjective:to evaluate Qingrehuoxue decoction reduces RA disease activity and radiological mobility progress evaluation.Methods:Enrolled active RA patients were randomized into in Qingrehuoxue(QRHX) group and Methotrexate(MTX) group and QRHX+MTX group. A24weeks of randomized, controlled study and cohort study to assess RA disease activity (DAS28score) improvement; and by taking medicine follow-up, used the Sharp/van der Heijde method to evaluate one-year hands X-ray score.Results:1. We enrolled in this study213patients which were entered in Full Analysis Set (FAS),129patients completed24weeks of treatment. The results show that three groups all can reduce disease activity and the mean decline of DAS28score of QRHX group, MTX group and QRHX+MTX group were1.91,1.97and1.29, QRHX group and QRHX+MTX group have better curative effect than MTX group.2. QRHX+MTX group can improve joint function just after one month treating, while QRHX group and MTX group improve joint function after three months treating; In improving joint function, QRHX+MTX group works faster, QRHX group is the second, and MTX group is the third. QRHX group and QRHX+MTX group have a better effect than MTX group to improving the RA joint function.3. Three groups have good effect to reduce the inflammatory laboratory index (ESR); QRHX+MTX group works better than MTX group. 4. Follow-up into radiology qualified cases progress evaluation of42cases, QRHX group and QRHX+MTX group of each21cases, two groups of Sharp total score, bone erosion score and the joint space narrow scores are no statistical significance. Two groups of Sharp scoring rate very similar, QRHX group0.24±0.28/0.19(0.04,0.29), QRHX+MTX group0.25±0.44/0.15(0.04,0.25). Two different extent the number of cases also similar (p=0.46), no radiation study progress of QRHX group in7cases,8cases of QRHX+MTX medicine; Radiology progress the obvious, QRHX group in3, the other in1.5. Safety, QRHX group adverse reaction rate (0%) lower than QRHX+MTX group (1.55%) and MTX group (11.4%). The safety of pure Chinese herbal medicine treatment compared with MTX or combined with MTX treatments are more reliable.Experimental Study1. In vivo experimental study:QRHX decoction inhibit the CIA rat Th/RANK pathways and bone destruction.Objects:Based on "osteoimmunology", through Th/RANKL pathways expounds QRHX decoction treatment RA possible "protect bone" mechanism.Methods:to cattle II collagen type SD rats induced arthritis models to MTX for the positive control medicine, QRHX water decoction and ethanol-extraction agent to fill the stomach six weeks. From an X-ray, HE dyeing, bone mineral density check more levels observed CIA rats inflammation and bone joint synovial hyperplasia damage change, ELISA test levels of CIA rats bone damage factor RANKL, IL-17, the MIP-1.Results:Serum levels of CIA rats RANKL, IL-17, the MIP-1of the normal group were significantly increased, QRHX water decoction and ethanol-extraction agent groups, serum levels of IL-17, the MIP-1were decreased significantly than model group, the RANKL level had decline trend. MTX group, serum levels RANKL, IL-17are decreased significantly than model group, the MIP-1had decline trend.2. In vitro experimental study:influence of QRHX decoction to RA synovial cell co-culture OPG, RANKL secretion, and broken bone cell differentiation Objective:to observe influence of QRHX decoction to RA synovial cell co-culture OPG, RANKL secretion, and broken bone cell differentiation role and discuss the molecular mechanism.Methods:to establish the in vitro human blood PBMC and normal synovial fibroblasts (RA-HFLS) of co-culture system, from healthy people in the peripheral blood separation get PBMC, using IL-1induction induction IL-17, and uses the ELISA, Western law and the TRAP active detection methods of QRHX decoction to observe the influence of the RAW264.7induction formulas RANKL broken bone cell differentiation and levels of OPG secretion, IL-17.Results:blood serum containing QRHX (5%,10%and20%) can dose dependent in vitro inhibition of the RA-HFLS proliferation, improve RA-HFLS cultivation in the level of OPG, and inhibit of RA-HFLS RANKL in the expression and reduce IL-17level. Conclusion1. Established the heat, dampness and blood stasis are the key pathogenesis and pathological of active RA bone destruction, Qingrehuoxue therapy is the basic principle.2. Clinical study confirms:Qingrehuoxue decoction can reduce RA disease activity, and delay RA bone destruction, has a "protect bone" role.3. Experimental study confirms:Qingrehuoxue decoction by inhibiting IL-17, cut down the RANKL expression, up the OPG level and adjusted the OPG/RANKL/RANK pathways, delay the bone destruction, confirm Qingrehuoxue decoction "protect bone" intrinsic mechanism. Characteristics and innovative points1. The theory innovation:first put forward heat, dampness and blood stasis are key points that lead to active RA inflammation lesion, bone destruction. Qingrehuoxue therapy can "protect bone" scientific hypothesis, enrich and perfect the Traditional Chinese Medicine for RA correspondence of prescription and syndrome, and correspondence of disease and syndrome therapy theory, clarify clinical Chinese medicine superior effect scientific basis.2. The idea innovation:2.1From intervention RA bone destruction, study the influence of Qingrehuoxue decoction treatment of RA patients bone destructive effects, to provide the influence of evidence-based medicine basis. 2.2Form" osteoimmunology " pespect, through the research on Th cell factors (IL-17), OPG and RANKL targets such as discussed Qingrehuoxue decoction invigorate the bone destruction systemic and local function mechanism, confirmed Qingrehuoxue decoction has "protect bone" scientific hypothesis.All in all, this project from clinical to the base, deeply researches on the Qingrehuoxue decoction treatment of active RA formulas blood disease curative effect and the influence of the RA bone destruction, and reveals the scientific connotation of Qingrehuoxue therapy treating active RA, guides clinical practice for Chinese medicine theory, and provides new methods and ideas of the scientific basis for Chinese medicine treatment of RA "protect bone" mechanism exploration.