Pyroptosis, a pattern of inflammatory death, is regulated by NLRP3-Caspase-1
inflammasome and GSDMD-FL
protein.
Antcin A is a small
triterpenoid molecule. In this study, Kupffer cells (KC) were used for in vitro model, which were treated with LPS and
Nigericin (L/N) to induce pyroptosis. ELISA was used to determine the influence of
Antcin A on the expression of inflammatory factors, IF was utilized to investigate NLRP3 and Caspase-1, PI staining was used to detect the opening level of membrane pores in KCs, C57BL/6J wild-type mice were fed with high-fat diet to construct a
NAFLD model, and were simultaneously treated with
Antcin A. H&E staining was used to detect hepatic pathological changes in mice, oil red staining was utilized to detect hepatic fat deposits in mice, IHC was used to detect the expression of NLRP3 and Caspase-1, Western blot was used to detect the expression levels of NLRP3
inflammasome (including NLRP3, ASC,
Caspase-1, GSDMD-FL and GSDMD-NT). Pull-down assay and immunoprecipitation assay were used to detect the binding between
Antcin A and NLRP3. As a result,
Antcin A could significantly inhibit the occurrence of pyrolysis, decrease the expression of inflammatory factors, inhibit the activation and assembly of NLRP3
inflammasome, and significantly down-regulate the expression of NLRP3, Caspase-1 and GSDMD-NT in KCs. In
NAFLD mice,
Antcin A could suppress the inflammatory response in liver tissues of mice, reduce
lipid deposition, down-regulate the levels of ALT and AST, and improve liver function in mice.
Antcin A could also inhibit the activation of NLRP3
inflammasome in liver tissue and decrease the level of inflammatory factors. In the study of mechanism, we revealed that
Antcin A could inhibit the assembly and activation of NLRP3
inflammasome by binding with NLRP3. In summary, in this study, we found that
Antcin A could inhibit pyroptosis in KC and alleviate the inflammatory response of liver tissue in
NAFLD by targeting NLRP3
inflammasome, which was one of the mechanisms of Anctin A in protecting liver.