Mechanism Of Hubei Fengyang Decoction Induced-Apoptosis Of Synovial Cells In CIA Rats

Objective To observe the therapeutic efficacy of Fengyang on rheumatoid arthritis and evaluate the effect of Fengyang to induce synovial fibroblast cell apoptosis in collagen-induced arthritic(CIA)rats.Methods Sixty healthy male SD rats with body weight 200�20g.Fifty rats were used to establish CIA rats model and randomly divided into CIA control control(CC)group,Tripterygium polyglycoside(TG)-treating group,and three three Fengyang-treating Fengyang-treating(FY)groups bred with low,medium and high dose of Fengyang decoction,10 rats in each group respectively.Another 10 normal rats as negative control(NC)group.From the 21 st day after the initial immunization,three FY rats were gavaged with maple Fengyang Decoction;TG group with Tripterygium glycoside suspension,and NC and CC groups with normal saline at the same volume.Rats in each group were gavaged for 28 consecutive days.During modeling and administration,knee joint diameter was measured and arthritis score was evaluate every 7 days.Rats were killed when finished 28 days observation.The artical tissues of rats were taken for observing the pathological changes of the arthritis and the expression of synovial TNFRII,and the expression of synovial TNFRII,Caspase-8,Bcl-xl proteins in different groups.And inflammatory factor in blood of rats.Results(1)General conditions: Compared with the negative control group,the weight of rats in the CC group increased slowly,the diameter of knee joint and the arthritis index increased significantly;compared with the model group,the weight of rats in each medication group increased significantly,the diameter of knee joint and the arthritis index decreased,among which,the effect of the high dose group of FY was better than that of the TG group(P<0.05).(2)Pathological observation of synovial tissue: in the CC group,synovial cells were increased,and the lining layer of synovial membrane was proliferated,thickened,disordered and evacuated;a large number of inflammatory cells infiltrated,and vascular hyperplasia and even pannus formed.Synovial tissue inthe treated group showed improvement in synovial cell proliferation,inflammatory cell infiltration,pannus and so on(P<0.05),while the improvement effect in the high-dose FY group was significantly stronger than that in the TG group(P<0.05).The results of HE staining showed that there was a large amount of inflammatory cell infiltration in the knee joints of rats in the model group.FY groups could significantly alleviate the swelling degre of knee joint,inhibit the swelling of foot,and decrease the score of HE staining microscopy(P<0.05).(3)ELISA results for cytokines:Compared with the model group,the levels of cytokines: Compared with the model group,the levels of TGF-? and IL-4 were significantly increased and the levels of IL-17 A and IFN-? were significantly decreased in each treatment group(P<0.05).(4)Western Blot test for apoptosis-associated proteins: Compared with the negative control group,the protein expression of TNFRII and Bcl-xl in knee synovium of CIA rats increased significantly,while the expression of Caspase-8decreased significantly(P<0.05).Compared with the CC group,the expression of TNFRII in TG group and low-dose FY group was significantly decreased;however which in medium-dose and high-dose FY groups was significantly increased(P<0.05).Compared with the CC group,the expression of Caspase-8 protein in each drug group was significantly increased;and compared with the TG group,this protein in low,medium and high-dose groups of FY groups was significantly increased(P<0.05).Compared with the CC group,the expression of Bcl-xl protein was decreased in all medication groups,among which there was a significant difference in the low-dose FY group and a significant decrease in the low-dose FY group compared with the TG group(P < 0.05).Conclusion The efficacy of Fengyang in treatment of collagen-induced arthritic rats may partially obtained by inducing synovial fibroblast cell apoptosis and regulating the expression of cytokines IL-17 A,TGF-?,IFN-?,IL-4.