白虎汤对蛋白泛素化与抗原递呈的影响OA

[Objective]By studying the ubiquitination of full length gasdermin D(GSDMD-FL)and Pro-caspase-1 in inflammatory cells and the interaction between the two proteins and major histocompatibility complex Ⅱ(MHC Ⅱ)molecules,the regulatory effect of the classic formula Baihu Tang on the endogenous antigen presentation process was investigated.[Method]Pyroptotic cell models were established in mouse alveolar macrophages(MHS)and myeloid-derived macrophages(BMDM),respectively.Pyroptosis was inhibited by Baihu Tang.The ubiquitination changes of endogenous antigens GSDMD-FL and Pro-caspase-1 in the cells were detected by co-immunoprecipitation technology.The expression of MHC Ⅰ and MHC Ⅱ proteins in cells before and after the intervention of Baihu Tang were detected by Western blotting.Through in vitro protein interaction studies,the effect of Baihu Tang intervention on the interactions between MHC Ⅱ and GSDMD-FL and Pro-caspase-1 was analyzed.[Result]In the pyroptosis models of MHS and BMDM,0.75-3 mg/mL of Baihu Tang significantly inhibited the release of lactate dehydrogenase(LDH)and interleukin-1β(IL-1β)in cells(P<0.05),and promoted the ubiquitination levels of pyroptosis-related proteins GSDMD-FL and Pro-caspase-1(P<0.05).Meanwhile,Baihu Tang also significantly promoted the expression of MHC Ⅱ in pyroptotic cells and inhibited the expression of MHC Ⅰ(P<0.05),with a dose-dependent manner.In vitro protein interaction results showed that Baihu Tang significantly enhanced the interaction between MHC Ⅱ and endogenous antigens GSDMD-FL or Pro-caspase-1,promoting antigen presentation.[Conclusion]The mechanism by which Baihu Tang promoted the ubiquitination-MHC Ⅱ presentation pathway of endogenous antigens was an important role in eliminating the risk factors of pyroptosis.