The Anti-inflammatory Effect Of Roburic Acid And The Related Mechanism

Objective:In this study,we investigated the anti-inflammatory effect and molecular mechanism of roburic acid in mast cells and macrophage.To provide a preliminary experimental basis and theoretical basis for the development of such compounds into anti-inflammatoriesMethods:1.The effect of roburic acid on cell viability of BMMC,HMC-1and RAW267.7 were tested by MTT assay.2.The release of histamine,production of IL-6 and LTC₄ were evaluated by enzyme-linked immunosorbent assay（ELISA）.The mobilization of Ca²⁺was evaluated by immumofluorescence method.3.The effect of roburic acid on NO production in LPS-stimulated RAW264.7 cells was investigated by using Greiss reagent.4.Expressions of cPLA₂,inducible nitric oxide synthases（iNOS）,NF-κB,IκB,IKK and MAPKs including ERK,JNK,p38 were determined by Western blot.5.PCA test was used to evaluate the anti-inflammatory effect of roburic acid in vivo.Results:1.In the BMMC,we found that roburic acid reduced the production of LTC₄,IL-6,and the release of histamine in IgE/Ag-induced BMMC.The phosphorylation of cPLA₂ was suppressed by roburic acid treatment.The phosphorylation of IκBαand IKKα/β,as well as translocation of NF-κB to the nucleus,was suppressed by roburic acid treatment in BMMC.In addition,phosphorylation of MAPKs including p38 was inhibited in BMMC,but the phosphorylation of JNK and ERK were not effected.In PCA test,roburic acid has shown reduce the amount of Evans blue in the ears and thickness of the ear.2.In the RAW264.7 macrophase,we found that roburic acid reduced LPS-induced production of NO and IL-6,and the expression of iNOS.Meanwhile,phosphorylation of IκBαand IKKα/β,as well as translocation of NF-κB to the nucleus,were suppressed by roburic acid treatment.In addition,phosphorylation of MAPKs including p38 and JNK were inhibited in RAW264.7.However,the phosphorylation of ERK was not effected.Conclusions:Roburic acid could reduce the release of histamine,production of IL-6and LTC₄.Roburic acid exhibited inhibitory activities on production of LTC₄ via blocking MAPKs pathway in BMMC;roburic acid also exhibited inhibitory activities on production of IL-6 via blocking IKK/IκB/NF-κB pathway in BMMC.Roburic acid also exhibited inhibitory activities on production of NO and IL-6 via blocking IKK/IκB/NF-κB and MAPKs pathway in RAW264.7,suggesting the potential application as a drug candidate for therapy of inflammatory diseases.