Skeletal muscle performance is influenced by both diet and the mode of exercise, with diet playing a crucial role in individuals' adaptation to exercise training. Our study investigated the interaction of oat bran (OB) diet and moderate intensity exercise training (MIET) on skeletal muscle function and athletic performance. Studies have reinforced the positive association of high-fat diet (HFD) with chronic systemic inflammation and corresponding peripheral skeletal muscle dysfunction during exercise training. OB could alleviate the inflammation, oxidative stress, and energy homeostasis disorder associated with HFD. We observed improvement in mice limb grip strength and endurance treadmill running distance with OB intake, accompanied by regulation of muscle function-related gene expression. OB intensified exercise training-induced carbohydrate and lipid metabolism, as indicated by changes in lactate, fumarate, malate, pyruvate, succinate, and citrate levels. Additionally, specific probiotic genera producing short-chain fatty acids (SCFAs) were promoted, while inflammation-related circulating metabolites were significantly decreased with oat bran intake. Our findings suggest interactions between OB and MIET improved HFD-induced skeletal muscle dysfunction on both the phenotype and the related mechanisms. This study is an extension of our previous study on the anti-fatigue effect of oat bran, providing a novel prospective by integrating exercise adaptation, gut microbiota, molecular metabolism and skeletal muscle in situ analysis.