Objective This study aims to assess the impact of a single bout of high-intensity exercise on thymus metabolism in mice and reveal the underlying molecular mechanisms.

Methods Male C57BL/6 mice were randomly assigned to exercise and control groups, and omics techniques were utilized to analyze the exercise-induced changes in the thymic metabolome and transcriptome.

Results The study identified 62 differentially expressed metabolites (DEMs), of which 55 were upregulated and 7 downregulated, with the DEMs mainly enriched in metabolic pathways such as protein digestion and absorption, tryptophan metabolism, and dopaminergic synapses. Additionally, 962 differentially expressed genes (DEGs) were identified, with 703 upregulated and 259 downregulated. KEGG enrichment analysis showed that the DEGs were mainly involved in several pathways, including the PI3K-Akt signaling pathway, cytokine-cytokine receptor interaction and oxidative phosphorylation. Integrated analysis revealed the significant enrichment of DEMs and DEGs in signaling pathways such as the TCA cycle and glycolysis. The qRT-PCR results were highly consistent with the RNA-seq data.

Conlusions A single bout of high-intensity exercise induced significant changes in the levels of metabolites such as docosapentaenoic acid, hydroquinone, indoxylsulfuric acid, and l-glycine in the thymus of mice. Genes such as Hsp90aa1, Pten, Bcl-2, FoxN1, FoxP3 and Sirt1 may play crucial roles in the immune regulation process.