EGCG Alleviates Skeletal Muscle Oxidative Damage in Heat-Stressed Pigs via Keap1/PGAM5 Complex-Mediated Mitophagy

The heat stress (HS) induced by high temperatures can result in oxidative damage to muscles, thereby compromising both muscle growth and immune function within the organism. Mitophagy serves as a pivotal pathway in alleviating excessive ROS production and subsequent oxidative damage. However, the potential role of epigallocatechin-3-gallate (EGCG), a natural antioxidant found in tea, in mitophagy under HS remains unexplored. Here, we present evidence of EGCG mitigating the oxidative-redox imbalance in porcine skeletal muscles induced by HS involving the antioxidant enzyme system mediated by the Keap1/Nrf2 pathway and mitophagy mediated by the PINK1/Parkin pathway. Importantly, we identified phosphate mutase 5 (PGAM5) for the first time as a key protein modulated by EGCG under HS conditions, regulating mitophagy. Inhibition of PGAM5 significantly attenuated the activation of mitophagy by EGCG. Molecular docking and dynamics simulations further suggested that EGCG directly binds to Keap1, disrupting the Keap1-PGAM5 protein interaction and thus promoting the release of PGAM5 and subsequently activating mitophagy. In summary, this study represents the first discovery of EGCG directly targeting Keap1/PGAM5-mediated mitophagy, which serves as a potential functional supplement for regulating the antioxidant capacity in pigs.