Galangin alleviates cold stress-induced splenic injury in Danzhou chickens by inhibiting NLRP3 inflammasome-mediated pyroptosis.

Cold stress poses a significant challenge to the poultry industry, leading to impaired health and economic losses. However, the specific mechanisms by which cold stress induces injury in the chicken spleen, a critical immune organ, remain poorly defined. This study aimed to investigate the protective effects of the natural flavonoid galangin against cold stress-induced splenic injury and to elucidate the underlying mechanisms, with a focus on the NLRP3 inflammasome-mediated pyroptosis pathway. Using an in vivo chicken model, we demonstrated that acute cold exposure (10±1°C for 48 h) significantly induced splenic structural damage, characterized by lymphoid depletion and necrotic debris. This was accompanied by a profound disruption of redox homeostasis, marked by increased malondialdehyde (MDA) and suppressed antioxidant enzyme activities, alongside robust activation of the NLRP3/caspase-1/GSDMD pyroptosis pathway and elevated levels of inflammatory cytokines (IL-1β, IL-18). Notably, prophylactic administration of galangin (50 mg/kg) effectively mitigated all these pathological changes in vivo, restoring antioxidant capacity, reducing inflammation, and inhibiting pyroptosis. In vitro studies using chicken HD11 macrophages confirmed that galangin suppressed cold stress-induced oxidative damage, inflammation, and pyroptosis. Critically, the protective effects of galangin were reversed upon NLRP3 overexpression, confirming that galangin acts, at least in part, by directly targeting this inflammasome. Molecular docking further predicted a strong binding affinity between galangin and the NLRP3 protein. In conclusion, this study identifies NLRP3-mediated pyroptosis as a key mechanism in cold stress-induced splenic injury and demonstrates that galangin alleviates this damage by inhibiting this pathway. These findings support the potential of galangin as a natural feed additive to enhance resilience against cold stress in poultry.