Objective To investigate the anti-fatigue efficacy of Hibiscus manihot L. and its potential mechanism, providing scientific basis and data support for the high-value utilization of Hibiscus manihot L. resources and the development of anti-fatigue functional foods.

Methods The anti-fatigue activity of Hibiscus manihot L. aqueous extract was evaluated using a weight-loaded forced swimming test in Kunming mice. The levels of blood lactic acid, hepatic glycogen, and muscle glycogen in mice were determined. The components of Hibiscus manihot L. were screened by network pharmacology, and their potential targets were predicted. The intersection of these targets and fatigue-related targets was obtained to construct a protein-protein interaction network. Gene ontology functional annotation and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were further performed to explore the potential anti-fatigue mechanism. Molecular docking was performed to verify the binding affinity between core components and key target proteins.

Results A total of 32 mice were included in this study. The aqueous extract of Hibiscus manihot L. significantly prolonged the exhaustive swimming time with load, reduced blood lactic acid levels, and increased the contents of hepatic glycogen and muscle glycogen in mice (P < 0.05). A total of 15 key components (mainly flavonoids, alkaloids, and amino acid derivatives) and 13 core targets were identified, which were mainly involved in biological processes such as aerobic oxidation electron chain transport, anaerobic glycolysis, and inflammatory response. Molecular docking results showed that core components such as chrysin had good binding activity with multiple core targets.

Conclusion Hibiscus manihot L. may exert anti-fatigue effects by regulating AKT1, HSP90AA1, and related pathways through flavonoids, alkaloids and other active components.

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