Study On The Mechanism Of Anti-gout Action Of Naru Sanwei Pills

Gouty arthritis(GA)is an arthropathy caused by the imbalance of uric acid metabolism in the body resulting in the deposition of sodium urate crystals(MSU)in the joints and tissues around the joints.The deposition of MSU crystals triggers the activation of the innate immune system,which triggers a strong inflammatory response in local joints and periarticular tissues.The disease is clinically characterized by recurrent episodes of severe pain,swelling,warmth,and redness in the gouty joints.Naru Sanwei Pills(Naru-3)is a compound medicine used by Mongolian medicine to treat rheumatism,joint pain,cold pain in waist and leg,toothache,diphtheria and other diseases.At present,western medicine clinically treats GA with allopurinol,colchicine,benzbromarone,febuxostat,non-steroidal anti-inflammatory drugs,and glucocorticoids.serious side effects.Traditional Chinese medicine compounds have the advantages of multi-component,multi-target,and few side effects,and most of them have the dual effects of lowering uric acid and anti-gout.With the current background of the modernization of traditional Chinese medicine,in-depth research on the material basis and mechanism of action of traditional Chinese medicine in the treatment of GA is imminent.Our research group found that Naru-3 has a significant curative effect on PO-induced hyperuricemia and MSU crystal-induced gouty arthritis.Therefore,modern technical means are used to clarify its related efficacy and potential therapeutic targets,and provide a theory for clinical application in accordance with.First,we identified the chemical constituents of Naru-3 and investigated the effect of Naru-3 on hyperuricemia(HUA)and its potential mechanism of action.A total of 31 compounds were identified based on Q-Orbitrap high-resolution LC/MS.The uric acid-lowering effect of Naru-3 was studied by establishing a hyperuricemia mouse model(intraperitoneal injection of potassium oxonate(PO),and detecting organ index,pathological changes of kidney,and renal function.In addition,the expressions of xanthine oxidase(XOD),adenosine deaminase(ADA),uric acid transporter protein and m RNA were also detected to explore the mechanism of Naru-3 reducing uric acid.The results showed that Naru-3 significantly decreased serum uric acid(Sur)levels in a dose-dependent manner,increased uric acid excretion rate(FEUA),and alleviated renal injury,indicating that Naru-3 had uric acid-lowering and renal protective effects.In addition,Naru-3 reduced the activity of hepatic XOD,down-regulated the expression levels of uric acid transporters GLUT9 and URAT1,and their m RNA levels,and up-regulated the protein and m RNA expression of uric acid-secreting protein OAT3.The results indicated that Naru-3 could reduce urate reabsorption and increase uric acid excretion to exert its uric acid-lowering and kidney-protecting effects.In conclusion,Naru-3 has uric acid-lowering and nephroprotective effects,and its mechanism may be a dual effect of reducing uric acid production and increasing uric acid excretion.Second,we explored the effect of Naru-3 on GA and its underlying mechanism.The GA mouse model was induced by joint injection of MSU crystals,and the toe swelling,hindlimb weight bearing,toe temperature,mechanical pain,thermal pain,inflammatory factors,and joint histopathological changes were detected to explore the anti-gout effect of Naru-3.The results showed that Naru-3 has anti-inflammatory and analgesic effects.Additionally,we examined joint NLRP3-related protein expression and validated using knockout mice.The results showed that Naru-3 could inhibit the activation of NLRP3.In conclusion,our study shows that Naru-3 exerts anti-gout effect by blocking the NLRP3/ASC/caspase-1 pathway,inhibiting the activation of NLRP3,and blocking the release of IL-1β.Thirdly,we explored the effect of the main compounds in Naru-3 on the air sac MSU inflammation model.By establishing a balloon model,MSU induced synovial inflammation,and detected changes in neovascularization,leukocyte count,inflammatory factors,and balloon histopathology.The experimental results show that the four monomers of piperine,piperamide,gallic acid and methyl gallate have anti-inflammatory effects.Finally,we also explored the effect of methyl gallate(MG)on hyperuricemic nephropathy(HN)and its underlying mechanisms.The HN model was established by intraperitoneal injection of PO,the improvement effect of MG on HN was evaluated,and the pathological changes of renal tissue,renal function,cytokines,and NLRP3-related proteins were detected.In addition,in vitro models of lipopolysaccharide(LPS)-stimulated bone marrow macrophages(BMDMs)and human peripheral blood mononuclear cells(PBMCs)were established to investigate the mechanism by which MG inhibits NLRP3 inflammasome activation.The results showed that MG significantly ameliorated HN by inhibiting uric acid production and promoting uric acid excretion and ameliorating NLRP3 activation-induced renal injury.MG,a direct NLRP3 inhibitor,inhibited NLRP3 inflammasome activation in macrophages,but had no effect on AIM2 or NLRC4 inflammasome activation.Mechanistically,MG inhibits NLRP3 inflammasome assembly by blocking NLRP3 oligomerization.Furthermore,MG was active against ATP-treated PBMCs and synovial monocytes from gout patients in vitro.In conclusion,MG has a renoprotective effect on PO-induced HN,and the mechanism may be related to blocking the oligomerization of NLRP3,thereby exerting its prominent anti-inflammatory activity in NLRP3-driven disease.In conclusion,the results of this research show that Naru-3 has uric acid-lowering,renal protection and anti-gout effects.The main mechanism may be to reduce uric acid production and increase uric acid excretion to reduce uric acid,and inhibit the activation of NLRP3 inflammasome to exert anti-gout effect.It provides a theoretical basis for the clinical application of Naru-3 against gout.