Dietary antioxidants alleviate antibiotic‐induced mitochondrial dysfunction through protein kinase AMP‐activated alpha (AMPKα) and nuclear factor, erythroid 2 like 2 (NRF2) pathway interaction

Background and Purpose Antibiotics and ibuprofen combinations cause mitochondrial toxicity and hepatotoxicity. This study investigated whether dietary antioxidants could protect against this damage via protein kinase AMP‐activated alpha (AMPKα)/ nuclear factor erythroid 2‐related factor 2 (NRF2) pathways. Experimental Approach Human umbilical vein endothelial cells (HUVECs) were treated with antibiotics (kanamycin, azithromycin, ampicillin or ciprofloxacin) plus ibuprofen with or without antioxidants. Azithromycin/ibuprofen induced hepatotoxicity was evaluated in C57BL/6J mice. Mitochondrial parameters including morphology, reactive oxygen species (ROS), mitochondrial membrane potential (Δ Ψ m ) and key proteins ( mitofusin 2 , AMPKα, glycogen synthase kinase 3 beta [GSK3B], NRF2 and haem oxygenase 1 [HO1]) were analysed. Key Results Antibiotics/ibuprofen combinations triggered mitochondrial fission, ROS overproduction and mitofusin 2 down‐regulation. Four antioxidants, that is, coniferaldehyde, raspberry ketone, gastrodin and eugenol, restored mitochondrial function and morphology. Coniferaldehyde and raspberry ketone effectively prevented in vivo hepatotoxicity and inflammation. Moreover, coniferaldehyde/raspberry ketone activated NRF2/HO1 while restoring AMPKα/GSK3B signalling. Conclusions and Implications Coniferaldehyde and raspberry ketone showed potent rescue effects in vitro against all antibiotic models and in vivo against azithromycin/ibuprofen‐induced hepatotoxicity through AMPKα‐GSK3B/NRF2‐HO1 modulation, with favourable safety profiles.