The Influence Of Yupingfeng Polysaccharide On T Lymphocyte Subgroup Of Adjuvant Arthrits Rat

Rheumatoid arthritis (rheumatoid arthritis,RA) is a systemic inflammatory autoimmune disease mainly involving the peripheral joints. It has features of progressive and recurrent attacks.There is no specific treatment. Mainly show is symmetrical, chronic,progressive polyarthritis.Recent studies indicate that the development and progression of RA is relate to various immune cells and their secretion of cytokines, of which T cell-mediated abnormal immune response is the main mechanism of rheumatoid arthritis.Particularly,CD4+T cells plays an important role in the pathogenesis of rheumatoid arthritis and participat stimulation and continuation of rheumatoid arthritis.There is obvious imbalance of cytokines secreted by Thl/Th2 cells in RA patient's serum and synovial fluid,and the imbalance is directly related to occurrence and outcome of RA.Thl cells mainly involved in cellular immune function.After stimulated by inflammatory cytokines,Thl cells secrete interleukin-2,interferon-y and so on in the RA process.IFN-y can activate macrophages, enhance expression of MHC-II antigen,enhanc activity of NK cell, inhibit fibroblast producted collagen,stimulate and enhance the inflammatory response.Th2 cells mainly produce IL-4, IL-10 and so on.IL-4 can reduce the secretion of IL-1 and TNF-α,inhibit corrosion and damage of arthrodial cartilage.Thl cells in the blood can slow the condition of RA patients while the major role of Th2 cells are protective.Therefore actively targeted immune therapy to restore the balance of immune cells, inhibit the activity of certain inflammatory mediators, regulation of the expression of related cytokines, may be a new way to control the chronic inflammation of RA.In this study,we extract Yupingfeng polysaccharide, used adjuvant arthritis rat induced by Freund's complete adjuvant to investigate the impact of T cell subsets and related cytokines in adjuvant arthritis rats.The main contents are organized as the following:1. Extract YPF-PYupingfeng herbs (Astragalus,Atractylodes macrocepha,Raidix Saposhnikoviae three components in accordance with the weight ratio of 3:1:1) 6.0 kg, with 75℃hot water boiling 2 minutes,each time 2 hours.Extraction liquid was concentrated to 20-30L, precipitate by alcohol,then centrifugation and then dried.What we get is Crude Yupingfeng polysaccharide.Crude Extractive is deqreased by ethyl acetate, then using sevage method to remove protein, the dried brown product named Yupingfeng polysaccharide (YPF-P).(yield is 0.85% of YPF weight).2. The induction and evaluation on AA modelIn addition to the normal group, rats of other groups are injected in Freund's complete adjuvant on rat's left toe.On the day before inflammation and the 12th,16th,20th,24th, 28th days after inflammation, we measured the right paw volume and arthritis index of rats.The results showed that YPF-P (40,80,160 mg-kg-1, ig) inhibite the secondary inflammation(secondary paw swelling, arthritis index) in AA rats.3. Effective of YPF-P on the immune regulation of AA ratsPeripneral blood lymphocyte proliferation response induced by concanavalin A (ConA) was examined with MTT assay. The results showed that AA rats on ConA-induced lymphocyte proliferation were significantly lower than the control group, YPF-P (80,160 mg-kg-1, ig) can improve the ConA-induced lymphocyte proliferation.It suggests that YPF-P regulat cellular immune function in rats with AA.4. Effective of YPF-P on T cell subsets of AA ratThe CD4+ and CD8+ T cell in peripheral blood were detected by flow cytometer.The results showed that the CD4+T cells of AA rat were significantly increased, while CD8+ T cells decreased.After treatment by YPF-P, the number of CD4+T cells significantly reduced and CD4+/CD8+ ratio lower.It indicate that YPF-P can restore the imbalance of T cell subsets.5. Effective of YPF-P on cytokines secret by AA rat T cellsELISA method was applied to determine the content of factor IFN-γand radio-immunity assay of factor IL-4.The results showed that 80 and 160 mg-kg-1 YPF-P can significantly reduce the content of IFN-y and slightly increased the content of IL-4.In addition, by testing IFN-y mRNA expression of lymphocyte with RT-PCR and analysis semi-quantitative.We found that each treatment group can reduce the expression of IFN-γmRNA.Suggest that YPF-P have certain effect on disordered cytokine network.All these results suggest that:YPF-P can reduce the secondary inflammation of AA rats.These effection may be related to the increases of lymphocyte proliferation response in peripheral blood and the adjustion of the balance of T lymphocyte subsets and related cytokines.