The Effects Of Bi Zheng Ning Decoction Combined With Methotrexate And Regulatory Of JAK-STAT Signaling Pathway On The Cold Dampness Type Of Collagen-induced Arthritis Rats

Objective:This project initially evaluate the effect of Bi Zheng Ning decoction on CIA model with animal experiments,further explore the efficacy of Bi Zheng Ning decoction and combined with methotrexate in the treatment of rheumatoid arthritis of the cold dampness type and targets based on JAK-STAT signaling pathway.To provide experimental basis for the clinical application of Bi Zheng Ning decoction and combined with methotrexate.Method:1.Forty Wistar rats were established a collagen-induced arthritis model and divided into 8 groups,then administrated by gavage with normal saline,the low?middle?high-dose of Bi Zheng Ning decoction,methotrexate and methotrexate combined with different doses of Bi Zheng Ning decoction for 4 weeks.The arthritis index score was observed weekly to initially evaluate the effect of Bi Zheng Ning decoction and the optimal ratio of Bi Zheng Ning decoction combined with methotrexate.2.Seventy Wistar rats were randomly divided into 7 groups,and each group except the normal were established a CIA model of the cold dampness type.Through the treatment of each dose of Bi Zheng Ning decoction ?methotrexate and combined with methotrexate on the CIA model to observe the influences of the rat body weight,joint swelling,detect inflammatory factors CRP,ESR content in serum and take synovial tissue to detect the levels of IL-6,MMP-3,MMP-9 by ELISA.3.Through the method of Western blot technology to explore the expression levels of SOCS1,SOCS3,PIAS3,P-STAT3 protein in synovial tissue,and using Realtime fluorescence quantitative PCR technology,researched the levels of JAK3 and STAT3 m RNA in synovial tissue.Results:1.After administrated for 2-4 weeks,each group of Bi Zheng Ning decoction,methotrexate group,and combined with methotrexate can reduce the arthritis index score(P <0.01),and methotrexate combined with the middle-dose of Bi Zheng Ning decoction was the lowest in the arthritis index score(P <0.01).The inhabitation of inflammation in the middle-dose of Bi Zheng Ning decoction was significantly better than the low-dose and high-dose groups(P<0.05,P <0.01).2.The middle-dose of Bi Zheng Ning decoction,methotrexate group,and the combination of Chinese medicine and western medicine all improved the body weight of CIA rats and reduced the livels of IL-6 in synovial tissue(P <0.01).Each administration group reduced the degree of joint swelling(P <0.01),decreased CRP and ESR content in serum(P <0.01),and the levels of MMP-3 and MMP-9in synovial tissue(P <0.01).The effect of the combination of Chinese medicine and western medicine was more significant(P<0.01).In the Bi Zheng Ning decoction groups,the middle-dose group was better than the low-dose and high-dose groups(P <0.05,P<0.01).3.The administration groups could promote the expression of SOCS1,SOCS3,PIAS3 protein in synovial tissue(P <0.01),and inhibit the expression of P-STAT3 protein(P <0.01),JAK3,STAT3 m RNA(P <0.01).In the administration groups,the combination of Chinese medicine and western medicine was significantly better than other administration groups(P <0.01).In the Bi Zheng Ning decoction groups,the middle-dose group was better than the low-dose and high-dose groups(P <0.05,P <0.01).Conclusion:Bi Zheng Ning decoction and combined with methotrexate in treating CIA model of cold and wet type can reduce arthritis index score,improve rat body weight,reduce joint swelling,reduce CRP,ESR content in serum and IL-6,MMP-3,MMP-9 level in the synovial tissue,and has a significant effect on inhibiting the inflammatory response.Its mechanism may be up the expression of SOCS1,SOCS3,PIAS3 proteins in synovial tissue,inhibit the overexpression of P-STAT3 protein and JAK3,STAT3 m RNA,thereby inhibiting JAK-STAT signaling pathway transduction and inhibiting the transcription of corresponding proinflammatory cytokines.Among them,the middle-dose Bi Zheng Ning decoction and combined with methotrexate have the best curative effect on CIA model of cold and wet type,which lays the experimental foundation for clinical application of rheumatoid arthritis.