| Objective:To investigate the pretection effects of soluble tumor necrosis factor (sTNFR)-Fc fusion protein on rats with collagen-induced arthritis(CIA) and To analyze TNF-αand OPN changes following therapy in order to understand their primary mechanism of interaction.Methods:CIA was induced by injecting bovineⅡcollagen(BⅡC),from the 13th day after first injection of BⅡC,the rats received sTNFR-Fc till the 36th day. Measured ankle joints anteroposterior diameters and weighs weekly; Pathology score of rat ankle joints were evaluated by HE stainin and toluidine blue staining;The blood plasma TNF-αand OPN levels were measured by ELISA and its histology expressd by immunohistochemistry.Take the synovial membrane of the normal control group and model group respectively to culture Fibroblasts synovial cells(FLS),Divided into control group, model group, model group + lipopolysaccharide (LPS), model group + lipopolysaccharide (LPS) + p38 channel inhibitor (SB203580),Then observing FLS s OPN mRNA change by RT-PCR.Further analysis the link of their and the possible signaling pathways.Rusults: In the modeling process, the normal control group s weight continued to grow, the average weight is 202.3 g before killed. Model group and treatment group incrase slowly, the average weight are 147.2 g and 194.3g before killed respectively. Model group s weight decreased to near baseline levels at 28 days. Comparing the three groups P <0.05, significantly different. HE staining can see severe synovial hyperplasia,a large number of plasma cells,lymphocytes and Toluidine blue dye staining showed severe loss of cartilage in model group than treatment group.Quantitative analysis showed pathological score in model group and treatment group treatment group scores were significantly reduced joint pathology[(8.16±1.02)vs(4.83±1.44),P<0.05]. ELISA results indicated mean plasma TNF-αand OPN values were 713.33±145.83 pg / ml, 4.34±0.56 ng / ml Respectively in model group, the treatment group were 67.62±20.05 pg / ml, 4.20±0.63 ng / ml, Between the two groups, serum TNF-αvalues were significantly different (P <0.05), while no significant difference between the value of plasma OPN (P = 0.688). sTNFR-Fc reduced the cells expressing OPN in the interface layer of synovium and cartilage(P<0.05). RT-PCR reveal that model by adding LPS stimulated cells was significantly increased OPNmRNA , By p38 inhibitor treatment could significantly reduce the formation of OPNmRNA, There were significant differences between the four groups (P <0.05)Conclusion: Pathology scores and ELISA results exhibit sTNFR-Fc had a good therapeutic action,It can significantly reduce the bone and cartilage damage of CIA mouse model , and can significantly reduce the expression of OPN in the sliding joints. Thereby delaying disease progression obviously. But It can not reduce the expression of OPN in the peripheral blood plasma. Willing to speculate the reasons for OPN might be involved in bone destruction and resorption rather than in the inflammatory process. Increased expression of TNF-αcan induce the expression of OPN, p38MAPK pathway play an important role in their... |
PDF Full Text Request