Study On The Material And Mechanism Of Jiu Wei Decoction Against Rheumatoid Arthritis

The main pathological feature of rheumatoid arthritis(RA)is synovitis,which is accompanied by synovial hyperplasia,cartilage and bone destruction caused by excessive synovial cell proliferation.At present,although the existing treatment drugs can effectively control the disease,they are expensive and have serious side effects.Traditional Chinese medicine has a long history in the treatment of RA,and has the advantages of good curative effect,low price and few side effects.However,due to the complex composition of traditional Chinese medicine,the active ingredients and mechanism are not clear,which limits the further development.Jiu Wei Decoction is a traditional Chinese medicine formulae composed of nine herbs.It is widely used in the folk to treat RA.However,there are few reports on the composition analysis and pharmacological efficacy of Jiu Wei Decoction.Based on this,this study intends to explore the pharmacodynamic material basis and mechanism of Jiu Wei Decoction in the treatment of RA through network pharmacology analysis,UPLC-MS/MS method and in vitro cell experiments.First,the pharmacodynamic substances and mechanism of action of Jiu Wei Decoction in the treatment of RA were preliminarily predicted by network pharmacology analysis.The results showed that TNF,IL-6,AKT,etc.may be the key targets of Jiu Wei Decoction in the treatment of RA;The anti-RA action mechanism of Jiu Wei Decoction may be achieved by regulating the expression of pathway proteins related to inflammation and cell proliferation.Secondly,6 active ingredients including quercetin,baicalein,wogonin,berberine,ferulic acid and kaempferol predicted by network pharmacology were analyzed and detected by UPLCMS/MS method.An analytical method for simultaneous detection and quantification of the six active components in the extract of Jiu Wei Decoction was established.Four components including quercetin,baicalein,wogonin and berberine were detected in the extract of Jiu Wei Decoction,and the quantitative analysis of the four components was carried out.Thirdly,the mechanism of Jiu Wei Decoction in the treatment of RA was explored through in vitro cell experiments.The results of western blotting experiments showed that Jiu Wei Decoction could reduce the phosphorylation level of AKT,increase the relative protein expression of Bax/Bcl-2,and effectively inhibit the proliferation of MH7 A cells;enzymelinked immunosorbent assay found that Jiu Wei Decoction could effectively inhibit the proliferation of MH7 A cells.It can inhibit the expression of inflammatory cytokines such as IL-6 and IL-1β in the supernatant of MH7 A cells induced by TNF-α,and has a good antiinflammatory effect.Finally,the anti-inflammatory effects of baicalein and wogonin in TNF-α-induced MH7 A cells were investigated in in vitro cell experiments.The enzyme-linked immunosorbent assay showed that both baicalein and wogonin could effectively reduce the contents of TNF-α,IL-6,IL-1β and other inflammatory cytokines in the supernatant of MH7 A cells induced by TNF-α.Comparing the effective threshold concentration of Jiu Wei Decoction extract and active ingredients,it was found that the efficacy of Jiu Wei Decoction was the result of the synergistic effect of low concentrations of active ingredients.In summary,the anti-RA effect of the traditional Chinese medicine compound Jiu Wei Decoction is to promote the apoptosis of synovial fibroblasts by regulating the expression of AKT,p-AKT,Bax,Bcl-2 and other proteins,and reduce TNF-α,which is achieved by the secretion of inflammatory factors such as IL-6 and IL-1β to inhibit inflammation.Quercetin,baicalein,wogonin,and berberine are the key active ingredients in Jiu Wei Decoction,which exert anti-inflammatory effects in MH7 A cells through synergistic effects.