Evaluation Of S1P Metabolism In The Synovial Membrane Of CIA Mice By Shexiangwulong Pill

Objectives 1.To evaluate the efficacy of shexiang-wulong pill in the treatment of CIA mice by comparing the changes of histopathological and histological synovial fluid indexes in collagen-induced arthritis(CIA)mice.2.To explore the possible mechanism of shexiang-wulong pill in the treatment of rheumatoid arthritis(RA)by comparing the effects of shexiang-wulong pill on the expression of sphingosine kinases(SphK1),sphingosine-1-phosphate(S1P)and s1 phosphate lymphoid(S1PL)in CIA mice.Methods 1.Among 45 DBA/1J mice,30 were randomly captured as the model group and the remaining 15 as the control group.The modeling process was divided into the first immune response and the second immune response on day 21.2 manufacturing model Mice in the modeling group were subcutaneously injected with 0.1 mL CII emulsion,which was made by the fusion of freund's complete adjuvant and bovine type II collagenic acetic acid solution in equal amounts.Making CIA models,when mice showed symptoms of secondary arthritis,made the pathological model successful.Group 3:15 mice were randomly selected from the successfully modeled mice as the treatment group and 15 as the model group.On the 22 nd day,the treatment group received musk Oolong suspended turbidity solution as gavage,while the control group and the model group received the same volume of normal saline.After continuous gavage treatment for 21 days,synovial fluid and synovial membrane were extracted on the 43 rd day.The levels of tumor necrosis factor(TNF-),interleukin-6(IL-6),1-acid sphenol(S1P),interferon-(IFN-)and serum S1 P in synovial fluid of mice were measured by double-antibody sandwich Elisa(Elisa).Western Blot and immunohistochemical staining were used to determine the expression levels of SphK1,S1 P and S1 PL in synovial tissue of the knee joints of mice.Results 1.Compared with the control group,the concentrations of TNF-?,il-6,IFN-? and S1 P in the synovial fluid of the CIA model group were significantly increased,while the concentrations of S1 P in the serum were decreased,with statistically significant differences(P < 0.05).The concentrations of TNF-?,il-6,IFN-?,and S1 P in synovial fluid of mice in the treatment group were significantly lower than those in the CIA model group,while the concentrations of S1 P in serum were increased,with statistically significant differences(P < 0.05)2.Western Blot results showed that the relative expressions of SphK1,S1 P and S1 PL proteins in synovial tissues of the CIA model group were significantly higher than those of the control group(P < 0.05).After treatment with shexiang wulong pill,the protein expressions of SphK1,S1 P and S1 PL in synovial tissues of CIA mice were significantly decreased(P < 0.05).3.Immunohistochemical results showed that the positive expression and average optical density of S1 P,SphK1 and S1 PL in synovial tissues of the CIA model group were significantly higher than that of the control group(P < 0.05).After treatment with shexiang wulong pill,the positive expressions of SphK1,S1 P and S1 PL and the average optical density of CIA mice in the synovial tissues of the knee joint were significantly reduced(P < 0.05).Conclusions 1 Musk oolong pills effectively reduced the inflammatory factors in synovial fluid of CIA mice.2.Musk oolong pill can inhibit the expression of S1 PL,SphK1 and S1 P in synovial membrane of knee joint.Figure5;Table1;Reference 187...