Copper (Cu), an environmental metallic pollutant, is closely associated with hepatic injury and inflammation, yet its molecular mechanisms remain incompletely understood. Although kaempferol (KAE) exhibits anti-inflammatory properties, its role in Cu exposure-induced hepatotoxicity, which is not fully clarified. In this study, we integrated animal experiments with network pharmacology and molecular docking to explore KAE's protective effects in a chicken model of Cu exposure-induced liver injury. Histopathological analysis revealed that Cu exposure induced inflammatory infiltration, hemorrhage, and collagen deposition, which were alleviated by KAE. Network pharmacology identified insulin (INS) as a key target, supported by strong binding affinity in molecular docking. Consequently, Cu exposure elevated blood glucose and upregulated INS and p38-MAPK expression, effects reversed by KAE. Further, the results showed that copper activated the p38-MAPK/NF-κB pathway, promoting NLRP3 inflammasome assembly and caspase-1-dependent pyroptosis, as evidenced by increased levels of NLRP3, ASC, caspase-1, GSDMD, N-GSDMD, IL-1β, and IL-18. Immunostaining confirmed upregulation of NLRP3 and N-GSDMD after Cu exposure, which KAE treatment suppressed. These findings elucidate a novel mechanism of Cu exposure-induced liver injury and highlight KAE's potential as a therapeutic agent for metal overload-related hepatotoxicity.

To Cite This Article: Guo S, Han M, Wu Z, Chen B, Yu J, Wang J, Huang H, Fouad D, Zhang Y, Tang Z, Zhang H and Guo J, 2026. Kaempferol alleviates copper exposure-induced liver injury in chickens by inhibiting pyroptosis. Pak Vet J, 46(4): 998-1007. http://dx.doi.org/10.29261/pakvetj/2026.073