Research On Therapy Of Reumatoid Arthritis By Shuangtengbitong Tincture

Objeetive:RA is a chronic, systemic autoimmune disease that is characterized by arthrosynovitis. Its main clinical manifestations are chronic, symmetrical, multi-joint synovitis and articular damage, and it is classified as an autoimmune inflammatory disease. The rate of incidence is higher among young adults, and it severely impair health due to the difficulty of treatment. RA is costly for individuals, health services and socio-economic impact in our society. However, due to the lack of efficacy and undesirable side effects related to many western medical treatments, so far effective and safe therapeutic treatment of RA is still desired.RA belongs to "Bizheng"."Lijie","Hexifeng" in traditional Chinese medicine. It is treated by traditional Chinese medicine syndrome differentiation and treatment under theory of traditional Chinese medicine. Traditional Chinese medicine play a pharmacodynamics through the multi-level, multi-link, multi-combination ways and target. They have advantages in anti-inflammatory and analgesic, regulating immune function. So Shuangtengbitong tincture was used to research its effects and investigate it mechanism of action.Methods:1. The anti-inflammatory activities of Shuangtengbitong tincture were oserved by utilizing the following models:carrageenin-induced paw edema in rats, xylene-induced ear edema, acetic acid-induced peritoneal capillary permeability and tampons-induced granuloma formation in mice.2. The analgesic effeets of Shuangtengbitong tincture were evaluated by hot-plate test and acetic acid-induced writhing response in mice as well as chlonic pain model in collagen-induced arthritic rats.3. Collagen-induced arthritic rat model was established to evaluate the anti-arthritic effetes of Shuangtengbitong tincture. We determined the change of primary paw swelling. Seeondary paw swelling, the arthritis scores and histological assessment of ankle articular synovium of all CIA rats. The levels of TNF-α, IL-1β and IL-6were measured by ELISA. Then the levels of the TNF-α、 IL-6. et al. in synovial tissues were examined by immunohistological staining, western blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis. 4. Synovial cells were isolated from CIA rats, then cultured. Effect of Shuangtengbitong tincture on synovial cells was observed, as well as the level of cytokines. such as IL-1βResults:1. Anti-inflammatory experiments:(1) xylene-induced ear edema experiment:the ear weigh of Shuangtengbitong tincture groups all were less than the model, and there were significant difference compared with the model group (P<0.05orP<0.01):(2) Cotton pellet granuloma formation experiment:granulation tissue weigh of Shuangtengbitong tincture groups all were less than the model, and there were significant difference compared with the model group (P<0.05or P<0.01);(3) Acetic acid induced capillary permeability in mice experiment:the OD value of Shuangtengbitong tincture groups all were less than the model, and there were significant difference compared with the model group in Shuangtengbitong tincture medial-dose and high-dose groups (P<0.05);(4) Carrageenin induced foot pad swelling experiment:foot pad swelling of Shuangtengbitong tincture groups all were less than the model, and there were significant difference compared with the model group only in Shuangtengbitong tincture high-dose groups at1h,2h,4h,6h:2. Analgesic experiments:(1) The hot plate experiment:the pain response time of Shuangtengbitong tincture groups were prolonged (P<0.05or P<0.01).(2) Chemical irritants experiment:the pain response time of Shuangtengbitong tincture medial-dose and high-dose groups were prolonged compared with the model group, and torsion times of mice within15mins was obviously reduced (P<0.05or P<0.01).3. Effect on CIA rats experiments:Aarthritis index and paw swelling of Shuangtengbitong tincture groups were less than the model group. And also improved the inflammatory response in synovial tissues. High-dose group had significant difference compared to the model group (P<0.05or P<0.01). The levels of TNF-α, IL-1β, IL-6, IL-8, et al. of Shuangtengbitong tincture groups in serum and synovial tissues were less than the model group (P<0.05or P<0.01). Meanwhile protein and mRNA expression levels of TNF-a. IL-6. IL-8. IL-10. TLR4. MyD88. NF-κB. IKκ. P-IκB were less than the model group. and high-dose group had significant difference compared to the model group (P<0.01). However protein and mRNA expression level of IκB was up-regulated and high-dose group had significant difference compared to the model group (P<0.01).4. Effect on synovial cells experiments:Shuangtengbitong tincture0.5and1mg/mL groups and MTX group decreased A value, and compared with control group.1mg/mL group and MTX group could obviously decreased A value (P<0.05or P<0.01).1mg/mL group and MTX group also reduced TNF-α、 IL-1β generation in LPS induced inflammatory response, improved IL-10level as well (P<0.05or P<0.01).Conclusions:1. Shuangtengbitong tincture could significantly inhibite the inerease in Paw edema induced by carrageenin. ear edema induced by xylene. vaseular permeability induced by acetic acid and granuloma induced by agar.2. Shuangtengbitong tincture obviously prolonged the writhing latent period and reduced writhing time in acetic acid-induced mice. Moreover. Shuangtengbitong tincture also increased the paw withdrawal threshold in CIA rats. However. the effect of Shuangtengbitong tincture on hot-plate pain threshold was not obvious.3. Shuangtengbitong tincture could remarkably inhibite the primary and secondary paw swelling, reduce the arthritis scores. And Shuangtengbitong tincture also significantly regulated inflammatory signal transduction networks and related factors, which manifested as reducing levels of serum TNF-α, IL-1β and IL-6. Meanwhile, the pathological examination demonstrated that synovial hyperplasia and infiammatory cells in filtration in CIA rats were suppressed by Shuangtengbitong tincture.4. Shuangtengbitong tincture inhibited synovial cells proliferation and reduced TNF-α、 IL-1β generation in LPS induced inflammatory response, improved IL-10level as well.