Combination Methotrexate,Leflunomide And Cyclophosphamide Intermitently Therapy On Arthro Phlogosisand Serum TNF-,IL-1(?) In Collagen Induced Arthritis Rats

BackgroundRheumatoid arthritis (RA) is a mutilation autoimmune disease. Cytokines play importanteffects on enlarging immunologic derangement and joints destroy. TNF-d IL-1(?) arerepresentative cytokines, who make joints chronic symmetry corrodibility change and vasculitis.Combination therapy is becoming increasingly popular in treatment of RA. However, the choiceof combining DMARDs can not be expected effective in daily practice. To expore newcombination therapy is very important. Based on successful combination therapy inleukocythemia, we first combined methotrexate with low dose cyclophosphamide to treat RApatients. MTX as prototype agent in RA is a cell cycle special stage agent, while CTX as a cellcycle nonspecial stage agent. During the last decade, the new combination therapy had beenproved more effective and safetive through we carried on clinicl practice.ObjectiveInvestigate the effeet of combination methotrexate(MTX), leflunomide （LEF） andcyelophosphamide(CTX) intermittently therapy on collagen-indueed arthritis rats.Investigate itspossible synergistic effect and evaluate the possible meehanisms through the arthritis index，arthritis swelling degree and the Serum TNF-, IL-1(?)Methods1. SD rats were raised, received two times mixed liquor of cattleⅡcollagen and Freund’scomplete adjuvant (CFA) by endermic injection. CIA rats were made.2. CIA rats were divided into8groups randomly, and received combination MTX, LEF andCTX treatment or other kinds control drugs respectively. Rats reeeived different treatments for6weeks.3. Observe the changes of arthritis index, arthritis swelling degree, and appearance ofimageology and pathology. Take blood, separate and obtain serum, then detected the serumTNF-, IL-1(?) by enzyme-linked immunosorbentassay. Results1. Arthritis index, arthritis swelling degree, and appearance of imageology and pathologyare labels,78rats who had received mixed liquor of cattle Ⅱcollagen and CFA happenedsymmetry polyarthritis, achievement ratio of model is92.3%.2. Arthritis index (AI): AI compared with model group of the therpy groups improvedsignificantly (P <0.05); The degree of AI improvement: the MTX combined with CTXgroup>the LEF combined with CTX group> the LEF combined with MTX group> the MTXgroup> the LEF group> the CTXgroup；AI improvement was better in combination group thanin monotherapy group (P <0.05)；In combination group, AI improvement was superior in MTXcombined with CTX group and in the LEF combined with CTX group than in the MTXcombined LEF group (P <0.05).3. X-ray radiography: X-ray changes of each therpy group was lighter than the model group,the combination groups X-ray efficacy were better than the monotherapy group, There was nosignificant difference between the combination therapy groups, the monotherapy groups werealso.4. HE joint pathological changes: synovial levels of the CIA model rats increased up to6-8layers, even more than10layers, disorganized, and inflammatory cell infiltration, cartilage, bonetissue is severely eroded.It was in turn reduce on the improvement in the degree of synovitis,cartilage and bone erosion in the MTX combined with LEF group,the MTX group, the LEFgroup, the CTX group,no significant differences among the groups; Synovitis, cartilage and boneerosion degree of improvement was significantly in the MTX combined CTX group and in theLEF combined CTX group than in the MTX combined LEF group、the MTX group、the LEFgroup and the CTX group (P <0.05).5. Combination MTX, LEF and CTX treatment have synergistic effect ininhibiting arthritisindex, arthritis swelling degree, and serum TNF-, IL-1(?) of CIA rats.(P<0.05).ConclusionsThis is the first research of effects of MTX combined with CTX intermittently according tocell cycle with CIA. Combination MTX and CTX therapy was more effective than single-agenttherapy. The results suggest combination cell cycle special stage agent MTX, LEF with cell cyclenonspecial stage agent CTX may be synergistic in the treatment of RA. MTX, LEF and CTX caninhibit TNF-IL-1(?). Action of TNF-inflammation stage ofCIA model. Inhibition elevated TNF-IL-1(?) level, maybe important mechanisms of combination MTX and CTX treatment according to cell cycle in treating RA.