Abstract: In this study, octacosanol (OCT) was evaluated for its ameliorative effect on exercise-induced fatigue in mice via behavioral and biochemical indices. The results showed that OCT significantly increased the rate of body mass gain, forelimb grip strength, exhaustive swimming time, total moving distance, and activity frequency in fatigued mice. Meanwhile, it markedly decreased the levels of blood lactic acid (BLA), lactate dehydrogenase (LDH) and malondialdehyde (MDA), and elevated the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Using network pharmacology, 327 potential anti-fatigue targets of OCT were identified, with 248 potential target proteins obtained from further analysis. Gene Ontology (GO) functional enrichment analysis identified 329 biological processes, 29 cellular components, and 67 molecular functions. Through network topological analysis, a total of 19 core targets were identified. Molecular docking results revealed that OCT had the highest binding affinity for the core anti-fatigue target proteins epidermal growth factor receptor (EGFR), nitric oxide synthase 3 (NOS3), protein kinase C alpha (PRKCA), and v-rel reticuloendotheliosis viral oncogene homolog A (RELA). Additionally, 16S rRNA sequencing analysis demonstrated that OCT had beneficial regulatory effects on the gut microbiota at the phylum, family, genus, and operational taxonomic units (OTUs) levels, significantly increasing the relative abundance of Lactobacillus, Alloprevotella, and Bacteroides. In conclusion, OCT may exert its anti-fatigue function by reshaping the gut microecosystem through a multi-target mechanism.

Key words: octacosanol; anti-fatigue; network pharmacology; gut microbiota