Phytochemical profile and hepatoprotective activity of Moringa oleifera Lam leaf extract in acetaminophen-challenged mice

Moringa oleifera leaves are rich in phytochemicals, including flavonoids and polyphenols, with reported antioxidant and hepatoprotective properties. This study evaluated the phytochemical profile of an aqueous-ethanolic extract and its efficacy against acetaminophen (APAP)-induced hepatotoxicity in mice. LC-MS/MS identified 80 metabolites, mainly flavonoids and amino acids. The extract showed moderate antioxidant activity in DPPH and FRAP assays, with a total phenolic and total flavonoid contents of 24.73 mg GAE/g DW and 2.95 mg CE/g DW, respectively. In vivo, mice given toxic APAP doses and treated with the extract (50 or 200 mg/kg) exhibited improved liver histology, reduced serum ALT and AST, suppressed CYP2E1 activity, and restored glutathione levels. Proinflammatory cytokines (IL-1α, IL-1β, IL-6, MIP-1α, MCP-1, TNF-α) and hepatic iNOS and NF-κB expression were also downregulated. Additionally, liver metabolomics revealed normalization of lipid metabolism and prostaglandin levels. These findings suggest that M. oleifera extract confers hepatoprotection via antioxidant, anti-inflammatory, and metabolic pathways, supporting its potential in functional food and nutraceutical applications. • Liquid chromatography-tandem mass spectrometry identified 80 metabolites in Moringa oleifera leaf extract • Extract reduced acetaminophen-induced liver damage in mice via multiple mechanisms • Hepatoprotection involved antioxidant, anti-inflammatory, and CYP2E1 suppression • Liver metabolomics showed normalization of lipid and prostaglandin profiles. • Extract demonstrated potent in vivo hepatoprotective effects in mice.