Observation Of Clinical Efficacy And Mechanism Of Action Of Juanbi Decoction In Treating Rheumatoid Arthritis

Objective:To explore the efficacy and safety of Juan Bi Tang(JBT)in the treatment of active rheumatoid arthritis through a multicenter,randomized,double-blind,placebocontrolled trial.In vitro and in vivo verification were performed using the rheumatoid arthritis fibroblast-likes synoviocytes and collagen-induced arthritis model.To explore the possible mechanism and target of anti-articular inflammation of JBT and its active components,and provide clinical and theoretical foundation for the popularization and application of JBT.Method:1.Study on the efficacy and safety of JBT in the treatment of active rheumatoid arthritis According to the multicenter,randomized,double-blind principle,115 patients with active rheumatoid arthritis(RA)who met the inclusion criteria were divided into JBT group and placebo group,and the two groups were treated with methotrexate(MTX)as the basic drug.After 12 weeks of clinical medication and 48 weeks follow-up,the two groups were observed for DAS28 score,EULAR score,CDAI score,pain swollen joint number,morning stiffness,SF-36 score,MOS score,serum inflammatory biomarkers,TCM syndrome integral.Evaluate the effectiveness and safety of JBT in the treatment of rheumatoid arthritis.2.Effect of JBT and effective components on the rheumatoid arthritis fibroblast-likes synoviocytes cellsHuman rheumatoid arthritis fibroblast-likes synoviocytes cells(MH7A)were selected as the research object.By observing JBT and 11 may have a monomer which contains anti-inflammatory effect on the influence of TNF-α-induced cytokine MH7 A and cytokines such as matrix metalloproteinase gene and protein levels.To evaluate the effects of sputum and its effective monomers on anti-inflammatory,anti-cartilage and bone destruction by regulating synovial cells,through the observation of the influence of intracellular signaling pathways,explore its possible mechanisms play a role.3.Effect of JBT and effective components on inflammation of collagen-induced arthritis miceEstablished mouse model of collagen-induced arthritis(CIA)and a threedimensional ultrasound assessment of CIA arthritis technology platform.The effects of JBT and active ingredients on joint inflammation in CIA mice were observed by arthritis clinical symptom score,3D-US evaluation,pathological score,Micro-CT bone destruction score,serum test,and immunohistochemistry.Thus,the efficacy and possible mechanism of action of the soup and effective monomers against antiinflammatory,anti-cartilage and bone destruction were evaluated.Results:1.JBT improves clinical symptoms and reduces disease activity in patients with active rheumatoid arthritisAfter 12 weeks of drug treatment and 48 weeks of follow-up,JBT can effectively reduce the DAS28 score of RA patients,reduce the activity of rheumatoid arthritis diseases,improve the EULAR standard of patients,and also reduce the CDAI score,reduce the number of joint pain swelling and morning.Stable time,improve the patient’s SF-36 quality of life score,improve patient sleep,reduce serum inflammation indicators,improve patients’ TCM symptoms,reduce TCM syndrome scores,and be safe,reliable,and have fewer adverse reactions.2.JBT and effective components inhibit TNF-α-induced inflammatory factors and MMPs production in MH7 A cellsThe appropriate drug concentrations were screened by CCK-8 method.It was found by PCR and ELISA that JBT and the effective components of nodakenin and astragalin can effectively inhibit the expression of inflammatory factors and MMPs genes and protein levels.The results of western blot and immunofluorescence showed that JBT and nodakenin could effectively inhibit the phosphorylation of p38,JNK,NF-κB p65 protein and NF-κB p65 nuclear translocation in MH7 A cells induced by TNF-α.Astragalin inhibits TNF-α-induced phosphorylation of p38,JNK,AP-1 c-Jun and p-cJun nuclear translocation in MH7 A cells.3.JBT and effective components improve joint inflammation in CIA mice and reduce cartilage and bone destructionA three-dimensional ultrasound and color Doppler technology platform for assessing joint inflammation in CIA mice was established.It was found that 3D-US can evaluate the synovial inflammation and vascular changes in the knee and ankle joints of mice in three dimensions.The 3D-US test results have a good correlation with the pathological scores.The 3D-US evaluation is superior to the clinical symptom scores.And MicroCT bone destruction score.By observing the clinical symptom score and the swelling thickness of the foot,it was found that JBT and the effective component astragalin can improve the clinical symptoms of CIA mice and reduce the swelling of the foot.Three-dimensional color Doppler ultrasound assessment,histopathological scoring,and Micro-CT bone destruction also showed that JBT and the effective component of astragalin can reduce the inflammation of the knee and ankle joint,synovial hyperplasia,cartilage and bone destruction.Serum detection and immunohistochemistry showed that the mechanism of its action may be related to reducing the production of serum inflammatory factors and inhibiting the expression of MMPs in synovial membrane and chondrocytes.Conclusion:1.JBT can effectively improve the disease activity of active RA patients,reduce the symptoms of arthritis,improve the quality of life of patients,and at the same time be safe,reliable and have fewer adverse reactions.2.JBT and its effective components,nodakenin and astragalin can effectively inhibit the production of inflammatory factors and MMPs in synovial cells.The mechanism of its action may be related to the inhibition of the phosphorylation of p38,JNK,NF-κB p65 and AP-1c-jun.3.JBT and the effective component,nodakenin and astragalin can improve joint inflammation and reduce cartilage and bone destruction in CIA mice.The mechanism of action may be related to reducing serum inflammatory factor production and inhibiting the expression of MMPs in synovial membrane and chondrocytes.4.3D-US can evaluate the synovial inflammation and vascular changes in the knee and ankle joints of CIA mice in vivo,non-invasively,three-dimensional and quantitatively.The 3D-US detection technique can be promoted in basic research of CIA mice.