The Investigation Of The Mechanism Of Expelling Wind And Removing Heat And Promoting Blood Circulation To Remove Meridian Obstruction Method In Treatment Of Collagen-induced Rat Arthritis

Objective: Establishment of expelling wind and removing heat and promoting blood circulation to remove meridian obstruction method.To evaluated the influence of JBY on pathological changes and inflammatory cytokines and metalloproteinase (MMP) levels in CIA Rats, exploring the possible mechanism for JBY in therapy of arthritis.Methods: (1)Theory study of expelling wind and removing heat and promoting blood circulation to remove meridian obstruction method. Chose 60 Wistar Rats to prepare type II collagen-induced arthritis (CIA) model, randomly divided into 6 groups, feed by different doses of JBY for 28 weeks. The arthritis scores and pathological arthritic changes were observed in both JBY-treated CIA rats and control ones. The levels of inflammatory cytokines interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and metalloproteinase MMP3 in CIA rat sera as well as in supernatant of cultured CIA rat synoviocytes treated with or without JBY were examined by ELISA. The influence of JBY on CIA synoviocyte proliferation were evaluated by MTT.Results:(1)CIA model results; the incidence of arthritis: 90% appears naked eye, joint swelling, mainly in the left hind legs, accounting for about 9/10, forelimb swelling accounted for 3/10. Serum ELISA test; a typeⅡcollagen antibody exists.(2)CIA rat joints pathological examination: a high degree of soft tissue lesions joint swelling, synovitis outstanding inflammatory response with a lining cell layer thickening, interstitial edema, and there is an abundance of inflammatory cell infiltration consisting mainly of lymphocytes, with pannus formation and invasion of cartilage and subchondral bone tissue caused by bone and cartilage destruction. Saline group: articular surface of the soft tissue inside the compartment can be seen the formation of a large number of spindle fiber cells. Organization sparse edema. And to see a large number of neutrophils and some lymphocytes, mononuclear cell infiltration. Capillary congestion significantly. And a red blood cell leakage. Medium dose Duocheng articular surface skin cells, spindle-shaped. The skin can be seen on a number of monocytes and lymphocytes. Some fibroblast proliferation. There capillary hyperplasia and congestion. Part of the plasma cell infiltration can be seen. Control group: tissue and cell arrangement rules, no significant inflammatory response.(3) serum inflammatory cytokines and metalloproteinases changes: After JBY treatment of serum TNF-α, IL-1β, MMP-3 an average drop of water to high and middle dose of the most significant (P <0.05)(4) synovial cell culture and stimulation: Each dose of JBY, were inhibited JBY trends in the growth of synovial cells.(5)JBY on the therapeutic effect of RA in rats: injection injection of typeⅡcollagen, Vladimir style is not entirely complete adjuvant, a mixture of acetic acid after 18 h, the right hind feet appear red, swollen, hot, continuous 3 d After gradually reducing. 10-14d,appearance of the multiple systemic arthritis, characterized by delayed hypersensitivity, 10 rat limb joints in 90% (56/60) there are different degrees of swelling, inflammation are numerous, persistent up to 2 more than a month Some developing joint deformity. In addition the majority of the ears and tail appeared in rat inflammatory nodules, some rats exhibited allergic corneal nebula and other extra-articular manifestations. Oral JBY rats in each group, joint lesions were improved, high-, medium-dose group was, significantly shorter duration (P <0.01). Fewer extra-articular manifestations, disease less. 28 days after treatment, the lesions from the general observation of both joints, such as pre-immune restoration, but joints slices on microscopic observation, the visible part of the joint organization of collagen in varying degrees of proliferation and periosteal plane. Conclusion: JBY was proved to be an efficient mixture in treatment of CIA rats. And the roles of inhibiting production of inflammatory cytokines and proliferation synoviocytes may be the therapeutic mechanism on treating CIA rats.