The Mechannism Study Of Total Saponins Of Clematis In The Treatment Of Rheumatoid Arthritis Which Based On JAK2/STAT3 Singal Pathway

Rheumatoid arthritis(RA) is systemic disease characterized by the proliferation and invasion of synovial tissues that leads to the destruction of bone and cartilage.Fibroblast-like synoviocytes(FLS) are central components of the aggressive, tumor-like synovial structure termed pannus which invades the joint space and cartilage. Multiple types of cells contribute to the pathological changes in rheumatoid synovium and accumulating evidence has implicated that fibroblast-like synovial cells(FLS) play a crucial role in both joint damage and the propagation of inflammation.In addition, the infiltration of inflammatory cells also attributed to the proliferation of RA-FLS and to the resistance of RA-FLS against apoptosis. As the resistance of RA-FLS against apoptosis has been identified as one of the key characteristics of these cells.JAK / STAT signaling pathway is one of the signal transduction pathways which was more extensive to be study in recent years. This pathway allows the extracellular chemical signals across the cell membrane and transmits the extracellular information to the gene promoter on the nuclear DNA, ultimately lead to the change of of DNA transcription and it’s activity in cell.JAK/STAT signal transduction pathway system is mainly mediated by cytokines, widely involved in many important biological processes,such as cell proliferation,differentiation, apoptosis and immune regulation etc. At present, researches on JAK / STAT signaling pathway as a target, and the corresponding drug has been developed for the treatment of RA.Therefore, further study about what role does JAK / STAT signaling pathway play in RA could help to reveal the pathogenesis of RA and to provide a basis for finding a safe and effective drug to therapy RA.Clematis is a traditional Chinese medicine which has been widely used to treat rheumatism, numbness, flexion and extension negative for many years. Total Saponins of Clematis(TSC) is an effective part extracted from Clematis, a previous study showed that it has anti-inflammatory analgesic and regulation of immune function. In this study,a rat model was established by using complete Freund’s adjuvant to induce arthritis.First,the overall level to investigate the effects of TSC on the role of adjuvant induced arthritis( AA) rats, then to explore the molecular mechanism of TSC inhibit RA-FLS molecular and cellular level in vitro experiments, observe the effect of TSC on the proliferation of RA-FLS and explore the regulatory of TSC on JAK2 / STAT3 signaling pathway in order to clarify the mechanism of the treatment of TSC on RA.The study includes the following two parts:1. The protective effect of TSC on AA rats which were induced by FCAFCA-induced arthritis rats model was established. The rats were randomly divided into normal control group, AA model group,TSC(50mg/kg、100 mg/kg、200 mg/kg),tripterygium glycosides tablet group(10 mg/kg). As shown with the results, TSC has a better protection for FCA-induced AA rats. TSC can effectively alleviate the weight loss in AA rats,significantly inhibit paw swelling of AA rats, reduce the joint score, change cell infiltration, reduce pannus production and lower the expression levels of IL-6 in serum, improve r the expression levels of IL-10.Meanwhile, TSC can effectively inhibit JAK2, STAT3 m RNA expression, decrease p-JAK2, p-STAT3 protein expression, while p-JAK2/JAK2,p-STAT3/ STAT3 relative expression may be reduced.2. Molecular mechanism of OM on anti-fibrosis2.1Inhibition effect of TSC on proliferation of RA-FLSRA-FLS cell model was established in vitro. TSC at a concentration of 0.5, 2.5,12.5, 62.5 and 312.5μg/ml could reduce the proliferation of RA-FLS; the inhibition rate was dose-depended related to the concentration of TSC.2.2 Influence of TSC on JAK2/STAT3 signaling pathway in RA-FLSResults showed that TSC could reduce JAK2, STAT3 m RNA relative expression,increase SOCS3, SHP-1m RNA relative expression while p-JAK2, p-STAT3 protein expression could be inhibited; at the same time, TSC also could reduce p-JAK2, p-STAT proteins relative expression and raise SOCS3, SHP-1 protein relative expression,suggesting that the effect of TSC on anti-RA can be achieved by regulating the JAK2 /STAT3 pathway.