Antioxidant Substance Basis and Mechanism of Flavonoids from Apios americana Medic.

To investigate the antioxidant material basis and mechanism of action of the flavonoid fraction (AMF) from Apios americana Medic., this study enriched the total flavonoids from Apios americana and evaluated their antioxidant activity using in vitro free radical scavenging assays and a zebrafish oxidative damage model. Additionally, qualitative analysis of the flavonoid components in Apios americana was conducted, and network pharmacology and molecular docking techniques were employed to explore the corresponding antioxidant targets and pathways. The results revealed that AMF exhibited potent DPPH and OH free radical scavenging abilities, with half-maximal inhibitory concentration (IC₅₀) values of 74.64 μg/mL and 22.54 μg/mL, respectively. Furthermore, in the zebrafish oxidative damage model, AMF significantly enhanced the superoxide dismutase and glutathione peroxidase activities, reduced the generation of reactive oxygen species, and mitigated overall cellular apoptosis in zebrafish. Qualitative analysis identified 15 flavonoid components in AMF, primarily consisting of genistein, apigenin, and daidzein. Further network pharmacology predictions indicated that AMF potentially targets 133 disease-related genes, with 45 core targets identified through PPI analysis. KEGG enrichment analysis suggested that AMF exerts its antioxidant effects through multiple signaling pathways, including PI3K-Akt, TP53, JUN, STAT3, TNF, and CASP3. Moreover, the docking results between AMF's active components and core targets were stable and consistent with the network pharmacology predictions. In conclusion, AMF demonstrates robust antioxidant activity, likely mediated by a synergistic interaction of multiple components, targets, and pathways. These findings provide a theoretical basis for developing and utilizing Apios americana as a functional food ingredient.