Ergothioneine alleviates exercise-induced fatigue associated with modulation of muscle transcriptome and gut microbiota

Abstract Fatigue often occurs after extended physical activity. A weighted exhaustive swimming-induced overtraining model was used to assess whether ergothioneine alleviates exercise-induced fatigue and is associated with changes in gut microbiota and fatigue-related gene transcription. We measured exercise endurance through swimming, grip strength, and rotarod tests, and evaluated locomotor activity via the open field and elevated plus maze tests. Oxidative stress and muscle injury were assessed using serum biochemical markers, and gut microbiota composition was analyzed by 16S rRNA gene sequencing. Overtrained mice showed decreased exercise endurance, increased levels of inflammatory cytokines, and altered gut microbiota. Ergothioneine supplementation at both low and high doses significantly improved endurance and reduced interleukin-6, blood urea nitrogen, and creatine kinase levels. It also restored gut microbiota composition by increasing Akkermansiaceae and Muribaculum while decreasing Desulfobacterota , Campylobacterota , and Erysipelotrichaceae . Transcriptomic analysis indicated that ergothioneine’s anti-fatigue effects were associated with changes in the AMPK and PPAR signaling pathways. These findings suggest that ergothioneine mitigates exercise-induced fatigue and may involve modulation of gut microbiota and inflammation- and metabolism-related pathways.