Hydrolyzed Ginsenosides of Panax notoginseng Flowers Attenuate Alcoholic Liver Disease by Inhibiting Oxidative Stress and Lipid Accumulation

Alcoholic liver disease (ALD) remains a major health burden with limited therapeutic options. While Panax notoginseng roots and leaves show hepatoprotection, the potential of P. notoginseng flowers (PNF) against ALD remains unclear. Notably, PNF exhibits solvent-dependent chemical profiles, with predominant ginsenosides undergoing significant hydrolysis at the C-3 position when extracted with water rather than methanol. This study evaluated the influence of ginsenoside hydrolysis on the protective efficacy of PNF against ALD by comparing water-extracted PNF saponins (WPNFS) and methanolic extracts (MPNFS). HPLC-UV characterized chemical profiles. In vitro models, including oleic acid-induced hepatic steatosis and H₂O₂-induced oxidative stress, were utilized to assess their bioactivity, while a chronic-binge ethanol feeding animal model was employed to evaluate in vivo efficacy. Network pharmacology and molecular docking identified potential targets, which were further validated through functional assays. Results indicated that WPNFS more effectively reduced lipid accumulation and mitochondrial ROS generation than MPNFS in vitro and attenuated ethanol-induced hepatic inflammation, oxidative stress, and hepatic steatosis in chronic-binge mouse models. Mechanistically, WPNFS activated the AMP-activated protein kinase (AMPK) pathway to promote lipid catabolism and upregulated the nuclear factor erythroid 2-related factor 2 (NRF2) pathway to enhance antioxidant defenses. Compound C (an AMPK inhibitor) or ML385 (an NRF2 inhibitor) abolished WPNFS benefits, confirming the involvement of these pathways. Structural analyses indicated that hydrolyzed ginsenosides in WPNFS enhance bioactivity. In conclusion, hydrolyzed ginsenosides from PNF mitigate alcohol-induced liver injury via dual activation of the AMPK and NRF2 pathways, offering a promising phytotherapeutic strategy for managing ALD.