The Effect Of Total Glucosides Of Paeony On IL-1β,IFN-γ In Blood Plasm And Synovium Of Rats With Adjuvant Arthritis

Objectives: Rheumatoid Arthritis (RA) is a kind of chronic, systemic, and autoimmune disease with key feature of arthrosynovitis, symmetric and destructive arthropathy. The disease progresses gradually and results in mutilation of joints. But the case of RA is still unclear. Many research have indicated that a lot of inflammatory cells and cytokines induced the abnormal cellular immunity and humoral immunity which conduced the systemic immunity and local joints dysfunction, all these are very important in the pathologic development of RA. Now the main therapeutic tool is the anti-rheumatic drugs, that can abatement symptom and step down the progression. But patients are often difficult to insist on taking medicines in long-term because of side effect or expensive cost. So it is imperative to find a kind of medicine with well match between price and effect. Total glucosides of paeony (TGP) is a kind of traditional Chinese drug, which used to cure autoimmune disease. It works gently, not only has the effect of anti-inflammation, adjusting immunity, but also has the effect of relief pain, and protecting the liver. It has been used in clinical therapy of rheumatic disease. But the relationship between dose and therapeutic effect is not clear. The aim of this study is to evaluate the therapeutic effect of TGP with different concentrations on AA rats induced by complete Freund's adjuvant from many aspects: weight, arthrocele, concentrations of IL-1βand IFN-γin blood and synovium, and investigate mechanism in treating RA.Methods: (1) The Wistar rats were divided into five groups in random. GroupⅠwas normal group; GroupⅡwas model group; GroupⅢwas model + low dose TGP group; GroupⅣwas model + high dose TGP group; GroupⅤwas model + MTX group. Rats of groupⅡ-Ⅴwere intradermal injected with 0.2ml complete Freund's adjuvant (CFA) on the root of tail. Fourteen days later, they were injected again with 0.1ml CFA to induce AA model. (2) On the 12th day after induction, the rats in groupⅢwere given an intragastric administration with TGP (50mg/kg·d); rats in groupⅣwere given an intragastric administration with TGP (300mg/kg·d); rats in groupⅤwere given an intragastric administration with MTX (2.7mg/kg·w). (3) The weight and volume of foot were detected and arthritic index (AI) was measured in different time. (4) The levels of IL-1βand IFN-γin serum were measured by ELISA on the 14th day and 28th day after cure. (5) Immunohistochemistry was used to detect the expression of IL-1βand IFN-γin synovium tissues and intensity was analyzed by computerized image system. (6) SPSS 11.5 statistical software was used to analyze the data. Results: (1) AI: In different time after induction, AI of groupⅡ-Ⅴwere significantly higher than that of groupⅠ(P<0.05), and the peak time was from the 20th day to the 25th day. On the 25th day, AI of groupⅢ, groupⅣwere significantly lower than that of groupⅡ(P<0.05),but similar with each other (P>0.05). On the 30th day, AI of groupⅤwas lower than that of groupⅡ(P<0.05) (2) Foot volume: Before induction, the foot volume of rats in every group was similar with each other. After 15 days, the foot volume of groupⅡ-Ⅴwere higher than that of groupⅠ(P<0.05).On the 20th day after induction, it reached the peak, and then it started to descend. Until 40th day, it still higher than normal (P<0.05). The curative effect of groupⅢand groupⅣemerged on the 20th day, five days earlier than groupⅤ. (3) Concentrations of IL-1βand IFN-γin blood plasma: On the 14th day of cure (the 26th day of induction), the concentrations of the two cytokines in groupⅡ-Ⅴwere obviously higher than groupⅠ(P<0.05).On the 28th day of cure (the 40th day of induction), they all descended, groupⅣand groupⅤdescended to normal, but groupⅡand groupⅢwere still higher than groupⅠ(P<0.05).Compared with groupⅡ, groupⅢ-Ⅴwere lower on the two days. GroupⅢ, groupⅣand groupⅤwere different with each other on the 14th day of cure. GroupⅣwas the lowest, and then was groupⅢ. On the 28th day of cure, groupⅣand groupⅤwere lower than groupⅢ, groupⅣwas similar with groupⅤ. (4) IOD of IL-1βand IFN-γin synovium: On the 14th day of cure, the IOD of the two cytokines in groupⅡ-Ⅴwere obviously higher than groupⅠ. On the 28th day of cure, they all descended, but still higher than groupⅠ(P<0.05). On the two days, the IOD of the two cytokines in groupⅢ-Ⅴwere lower than groupⅡ. On the 14th day of cure, the IOD of the cytokines in groupⅢ, groupⅣ, groupⅤwere different with each other. GroupⅣwas the lowest, and then was groupⅢ, groupⅤwas the highest of the three. On the 28th day of cure, the IOD of the cytokines in groupⅣand groupⅤwere lower than groupⅢ(P<0.05), that in groupⅣwas similar with that in groupⅤ(P>0.05). (5) Analyze of dependability: There were significant linear direct correlation between AI with foot volume, IL-1βwith IFN-γin plasma, IL-1βwith IFN-γin synovium. And the two cytokines not only in plasma but also in synovium respectively and directly correlated with foot volume.Conclusions: (1) AI was significantly linear direct correlated with foot volume. They all could reflect the swelling of joint, but the latter was better in precise and objective. (2) The levels of IL-1βand IFN-γin blood plasm or in synovium were obviously increased after induction and significantly correlated with foot volume, hinting that these two cytokines educed important effect in pathological process of RA. (3) There was a significant linear direct correlation between IL-1βwith IFN-γin blood plasma or in synovium, and both of them influenced each other, forming a vicious network in pathological process of RA. (4) TGP could lighten symptom by descending the concentrations of IL-1βand IFN-γin blood plasm. (5) TGP could reduce the expression of IL-1βand IFN-γin synovial tissues, lighten swelling of synovial cell, reduce infiltration of inflammatory cell, and inhibit hyperplasia of synovium. (6) The therapeutic effect of TGP was earlier than MTX. On the 14th day of cure, the effect of TGP was better than MTX; On the 28th day of cure, the effect of large dose TGP was similar with MTX.