Effect Of Tripterygium Glycoside With Sodium Ferulate To The Rats With Collagen Induced Arthritis And The Expression Of RANKL/OPG In Synovium

Objective: Rheumatoid Arthritis (RA) is a kind of chronic and autoimmune disease. Its diseased region is joint. It shows that articular synovium tissues abnormal hyperplasy, lots of inflammatory cells invasion and progressive destruction of bone and cartilage in the joint of RA. The destruction of bone and cartilage is the major reason to disability in RA. Tripterygium Glycoside is isolated from Tripterygium wilfordii Hook.f, which is used in immune suppression and anti-inflammatory effect as the best activity of chemical compound. Sodium Ferulate is isolated from Chinese Angelica and Szechwan Lovage Rhizome which is used in improving circulation, anti-oxidation. We make collagen induced arthritis (CIA) of rats and research the level of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), nitric oxide (NO), induced nitric oxide synthases (iNOS) in the serum and the expression of receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) in the synovium to discuss therapeutic effect and bone protection mechanism of traditional Chinese drug effective constituent compatibility and supply evidence to clinical application.Methods: Choose Wistar rats weight from 130g to 150g, the number of the male and female is egual. Choose 8 rats as normal group and the others are used to make model of CIA. Inject 0.2ml chicken typeⅡcollagen solution to the tail, right hind limb and back together, and boosted at the 14th day. Divid the models of CIA randomly into model group, TG group, SF group and TG+SF group at the 21th day. There are 8 rats in every group. Except model group, give the other groups medicines once a day and keep 28 days. Observe its general status, measure weight, paw swell thickness and assess arthritis index (AI) before making model, giving medicine and after drug withdrawal. After drug withdrawal, the rats are sacrificed by drawing blood from abdominal aorta. Then get and hold the serum. Get and fix the ankle joints. The levels of TNF-αand IL-1βin the serum were measured by radioimmunoassay. The levels of NO in the serum were measured by nitrate reductase assay. Measure the level of iNOS. The ankle joints were dehydrated, wrapped, sliced. HE staining was done to observe the pathologic change by light microscope and to assess the pathological score. Immunohistochemistry was used to detect the expression of RANKL and OPG in synovium tissue and their intensity were analyzed by computerized image system. All the data is analyzed by SPSS13.0 statistical software and expressed as x±s. The comparison of mean among groups is evaluated by one-way analysis of variance (ANOVA).Results:1 The effect of TG&SF to general condition of CIA rats The rats of CIA appeared swelling in the 10th day post-modal, achieved peak in the 14th day, and obviously after booster. The weights of model rats decreased in the 21th day of post-model, there was significant difference compared to normal group (P<0.05). Gradually, AI and claw swell thickness increased and achieved peak in the 28th day. There was significant difference compared to normal group (P<0.05). After administrated for 28 days, those of medication groups decreased, and there was significant difference compared to model group (P<0.05); compatibility group was significantly lower TG group and SF group (P<0.05). There was significant difference between model and normal group (P<0.01).2 The effect of TG&SF to level of TNF-αand IL-1βin the serum of CIA ratsThe levels of TNF-αand IL-1βin the serum of model group and medication groups was significantly higher than normal group's (P<0.05). The model group's and SF group's was significantly higher than TG group's and compatibility group's (P<0.05). There was no significant difference between model group's and SF group's (P>0.05).3 The effect of TG&SF to level of NO and iNOS in the serum of CIA ratsThe levels of NO and iNOS in the serum of model group, TG group and SF group was significantly higher than normal group's and compatibility group's (P<0.05). The model group's was significantly higher than TG group's and SF group's (P<0.05). There was no significant difference between TG group's and SF group's (P>0.05). There was no significant difference between compatibility group's and normal group's (P>0.05).4 The effect of TG&SF to pathologicals result in the joint tissue of CIA ratsPathologicals results showed that there were a lot of macrophages and T cells in synovial tissues, articular surface didn't complete and pannus emerged in model group; there were less macrophages and T cells in synovial tissues and bone destruction degree decreased. Remarks of model group and medication groups were higher than those of normal group (P<0.05); Remarks of model group were higher than those of medication groups (P<0.05); Remarks of TG group and SF group were higher than those of compatibility group (P<0.05); There was no significant difference between TG group's and SF group's (P>0.05).5 The effect of TG&SF to expression of RANKL and OPG in the synovium tissue of CIA ratsImmunohistochemistry showed that the expression and ratio of RANKL and OPG in synovium of model group and medication groups was higher than those of normal group (P<0.05); those of model group and SF group were higher than those of TG group and compatibility group (P<0.01); those of TG group were higher than those of compatibility group (P<0.05). There was no significant difference between model group's and SF group's (P>0.05).Conclusions:1 Tripterygium Glycoside combined with Sodium Ferulate has notable therapeutical effect on rats of CIA. Its is better than signal Tripterygium Glycoside's. This shows that Sodium Ferulate has synergistic effect to Tripterygium Glycoside therapying CIA.2 The effective mechanism that Tripterygium Glycoside has a good therapeutic effect on rats of CIA may correlate with decreasing the level of TNF-α, IL-1β, NO and iNOS in serum and the expression of RANKL in synovium.3 The synergism mechanism of Sodium Ferulate towards Tripterygium Glycoside may include that it could decrease the level of NO and iNOS, so lessen bone destruction; it could modify blood stasis symptoms, so enhance Tripterygium Glycoside's biology utilization.4 The synergism of traditional Chinese medicine effective ingredient compatibility to CIA supplys a new idea to treating RA by integrated traditional and western medicine.