Experimental And Clinical Study On Effect Of Rheumatoid Arthritis Treated By Long Yi Qing Feng Mixture

Objective:(1) To observe the clinical effect on rheumatoid arthritis (RA) treated by long yi qing feng mixture(LYQF).(2) To investigate the changes on the level of TNF-a and IL-6 in serum and the changes on the distribution of T-cell subsets in peripheral blood in patient with RA; And to investigate the influence on the expression of TNF-a and IL-6 in joint tissue and the influence on the expression of TNF-amRNA、IL-6mRNA in synovium of CIA rats treated by LYQF.(3) To contrast the effectiveness between LYQF ultramicro-powder (LYQFu) and LYQF decoction (LYQFd).Methods:(1) 60 case wind-damp-heat Bi RA patients were randomly divided into treatment group and control group, there were treatment group treated with LYQF mixture and control group treated with methotrexate (MTX). Observe the changes of joint’s tender and swollen, and the disease activity score (DAS) before and after treatment. Test the blood count, stool, urine, function of liver and kidney, ESR, CRP, IgM were examined before and after treatment separately.(2) Analyze the changes on level of TNF-a and IL-6, and the distribution of T-cell subsets in peripheral blood in patient with RA by ELISA and tri-colour fluorescence labeled monoclonal antibody and using multiparameter flow cytometry.(3) The experiment arthritis rat model was established by injected BovineⅡcollagen(BⅡC)and complete Freunds adjuvant(CFA).The CIA rats were divided into model group, LYQFu group, LYQFd group, Tripterygium wilfordii Hook(TWH) group, and blank-controlled group randomly. Rats in Model group (n=9) were given saline by perfusion of stomach, LYQFu group(n=10) were given LYQF ultramicro-powder by perfusion of stomach, LYQFd group (n=9)were given LYQF decoction by perfusion of stomach, and TWH group(n=9)were given Tripterygium wilfordii multi-glucoside by perfusion of stomach, blank-controlled group were drink water freely. All the rats were killed after the treatment 28 days.(4) Observe the changes of weight and arthritis index score of CIA rat’s during in treatment and investigate the Pathological changes in joint’s tissue of CIA rats.(5) Examine the expression of TNF-a and IL-6 in CIA rat’s joint tissue and TNF-amRNA AND IL-6mRNA in CIA rat’s synovium by immunohistochemistry (IHC) and Real-time PCR.Result:(1) This clinical study indicate that LYQF decoction have reliable effectiveness on patient with RA, which compared with MTX, P>0.05.(2) LYQF decoction inhibit the level of TNF-a and IL-6 in peripheral blood of patient with RA, which compared with pretherapy, the difference is obvious, P<0.01. LYQF decoction can regulate the distribution of T-cell subsets, which down-regulate the count of CD4+T cell and up-regulate the count of CD8+T cell, compared with pretherapy, difference is obvious, P<0.05.(3) LYQF ultramicro-powder and LYQF decoction both meliorate the weight of CIA rats, compared with Tripterygium wilfordii multi-glucoside, P<0.05; LYQF ultramicro-powder improve the arthritis index in CIA rats obviously, compared with LYQF decoction, P<0.05.(4) Pathological observations:Model group:synovium surface roughness, synovial cell hyperplasia, fibrous tissue in synovial tissue hyperplasia, the blood capillary congestive expansion, a large number of inflammatory cells and fibroblast infiltration can be seen, some of synovial tissue edema.LYQFu group:the layer of synovial cells uneven, but the majority of cells is 1-4 layers and only see synovial cells which mild hyperplasia, synovial tissue can also see the hyperplasia of blood capillary and fibrous tissue, there is a small amount of inflammatory cell infiltration.LYQFd group:synovial surface roughness, synovial cell hyperplasia, some region of synovial tissue hyperplasia, part region of the capillary increased, there is inflammatory cell infiltration.TWH group:the majority of synovial surface smooth, but synovial tissues can be seen villiform hyperplasia, some synovial tissue is still seen increased capillaries, and the expansion hyperemia, a small amount of inflammatory cell infiltration.(5) LYQF ultramicro-powder and LYQF decoction both inhibite the expression of TNF-a、IL-6 in the joint tissue and in the synovium of CIA rats obviously, compared with model group P<0.01. and LYQF ultramicro-powder is better than LYQF decoction significantly, P<0.05.Conclusion:1. LYQF decoction which improve the value of Hb and count of RBC obviously have reliable effectiveness on patient with RA than with MTX.2. LYQF decoction can regulate the imbalance condition in T-cell subsets distribution in patient with RA.3. LYQF decoction can inhibit the level of TNF-a、IL-6 in peripheral blood in patient with RA.4. LYQF decoction also can inhibit the level of TNF-a、IL-6 in rat’t joint tissue.5. LYQF decoction can down-regulate the expression of TNF-amRNA and IL-6mRNA in the rat’t synovium.6. The effectiveness of LYQF ultramicro-powder was significantly better than it’s decoction.