Astilbin attenuated pyroptosis caused by terbuthylazine-induced histone lactylation in grass carp hepatocytes

Terbuthylazine (TER), a triazine herbicide, can trigger excessive production of reactive oxygen species (ROS) in organisms, leading to liver damage in aquatic species. Under conditions of abnormally elevated ROS, increased glycolysis promotes the lactic acid accumulation, which in turn enhances histone lactylation and ultimately triggers pyroptosis. Meanwhile, N6-methyladenosine (m 6 A) methylation contributes to the pathogenesis of various diseases by regulating redox states. Astilbin (AST), a flavonoid with powerful antioxidant and anti-inflammatory properties, offers effective protection against injury in organisms. However, whether AST can attenuate TER-induced liver injury and the underlying mechanism remain unclear. Therefore, we treated L8824 cells with siMETTL14, NAC, PX-478, 2-DG, DMOG or Oxamate to achieve it. By integrating network pharmacology with bioinformatics, we found that AST participates in m 6 A methylation, thereby modulating redox homeostasis, glycolysis process, and pyroptosis. Following experimental verification, AST treatment significantly reduced FOXO1 m 6 A methylation, leading to decreased expression of HIF-1α and factors related to glycolysis process, histone lactylation, and pyroptosis. Conversely, it effectively promoted the expression of factors related to oxidative stress upon TER exposure. Flow cytometry and ROS staining further demonstrated that AST suppressed excessive ROS production. Molecular docking analysis and CETSA revealed a binding interaction between AST and METTL14. In summary, this study revealed that AST inhibits METTL14-mediated m 6 A methylation of FOXO1 to maintain redox homeostasis, thereby suppressing histone lactylation and ultimately attenuating pyroptosis in L8824 cells.