Integrated network pharmacology and metabolomics to explore the mechanism of Kadsura coccinea root in alleviating acetaminophen-induced liver injury.

ETHNOPHARMACOLOGICAL RELEVANCE: The Kadsura coccinea (Lem.) A. C. Smith is known as "Heilaohu" of the Tujia ethnomedicine in China. It demonstrates activities such as antioxidant properties and liver protection, making it suitable for the treatment of rheumatoid arthritis, as well as liver and gastrointestinal diseases. In recent years, research on K. coccinea has mainly focused on the analysis of chemical composition and screening for activity, however, the research on the mechanisms of liver protection has not yet been addressed.

AIM OF THE STUDY: This study aimed to investigate the hepatoprotective mechanisms of Kadsura coccinea roots ethanol extract (KCE) against Acetaminophen (APAP)-induced acute liver injury (ALI).

MATERIALS AND

METHODS

APAP-induced ALI was induced in male C57BL/6J mice through intraperitoneal APAP injection, followed by 5-day oral KCE treatment. Integrative approaches, including network pharmacology, molecular docking, untargeted metabolomics, Western blot, hematoxylin-eosin staining, and immunohistochemistry, were used to determine KCE's therapeutic effects.

RESULTS

Network pharmacology identified 672 differentially expressed genes, with 128 overlapping targets potentially mediating KCE's hepatoprotection. Molecular docking showed that he binding energy of isovaleroylbinankadsurin A with the top 10 proteins are range from -9.6 to -5.0 kcal/mol. Gene ontology and Kyoto encyclopedia of genes and genomes analyses revealed that KCE alleviates liver injury by modulating hepatic metabolic pathways. Untargeted metabolomics identified 381 upregulated and 960 downregulated metabolites in the model group versus controls. Additionally, the high-dose KCE group exhibited 280 upregulated and 237 downregulated metabolites versus the model group. Pathway enrichment analysis exhibited significant associations with the synthesis and degradation of ketone bodies, phenylalanine-tyrosine and tryptophan biosynthesis, riboflavin metabolism, glycine-serine and threonine metabolism, glyoxylate and dicarboxylate metabolism, ascorbate and aldarate metabolism, glycerophospholipid metabolism, and arachidonic acid metabolism. This indicated that KCE may alleviate liver injury by regulating apoptosis- and oxidative stress-related pathways. In vivo studies confirmed that KCE reduced serum alanine aminotransferase and aspartate aminotransferase levels, elevated glutathione, ameliorated histopathological damage, suppressed apoptosis and oxidative stress, upregulated Bcl-2/BAX ratio, HO-1, and Nrf2 expression, and downregulated p-Nrf2, Keap1 and Cleaved caspase 3 level.

CONCLUSION

This study determines KCE's efficacy as a hepatoprotective agent by interacting network pharmacology, untargeted metabolomics, and experimental validation, which demonstrates KCE. In terms of further research, KCE attenuates APAP-induced oxidative stress and apoptosis by inhibiting the Nrf2-Keap1 signaling axis, providing evidence for its potential to prevent APAP-induced ALI.