In vitro‐Digested Cervus nippon Velvet Antler Polypeptides Alleviate Alcoholic Liver Disease by Suppressing CYP2E1 Activity and Enhancing AMPK Signaling Pathway

Deer antler velvet (DAV), historically documented in Shennong's Classic of Materia Medica, is traditionally valued as a functional food believed to promote liver and kidney health and to strengthen the musculoskeletal system. Modern research highlights the bioactivity of sika deer (Cervus nippon Temminck) velvet antler peptides (sVAP), though their potential against alcoholic liver disease (ALD) remains unclear. This study aims to prepare sVAP through in vitro digestion and systematically evaluate their potential protective effects against ALD, while further elucidating the underlying mechanisms of action. Our findings demonstrate that the 1-3 kDa fraction (sVAP-1-3 kDa) exhibited the strongest hepatoprotection, reducing alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), total cholesterol (TC), and malondialdehyde (MDA) while enhancing superoxide dismutase (SOD) and glutathione (GSH). It alleviated reactive oxygen species (ROS) accumulation, suppressed apoptosis and lipid deposition, and activated nuclear factor erythroid 2-related factor 2 (Nrf2) signaling. Network pharmacology predicted 430 targets enriched in cytochrome P450 metabolism, lipid regulation, and inflammation. Docking showed strong binding of CYP2E1 to peptide motifs such as DFGFDGD, YFPH, FPAVP, and TYFPH. Western blotting confirmed that sVAP-1-3 kDa modulated CYP2E1/Nrf2/HO-1 and AMPK pathways, downregulating CYP2E1, SREBP-1c, FAS and PPARγ while upregulating Nrf2, HO-1, p-AMPKα. This study employed simulated in vitro digestion to prepare sVAP first. sVAP-1-3 kDa exerts antioxidative and anti-lipogenic effects, thereby attenuating oxidative stress, lipid synthesis, and ALD progression, providing pharmacological evidence for the traditional use of DAV. PRACTICAL APPLICATIONS: Polypeptides extracted from antler velvet, a substance with both medicinal and dietary value, have been shown to exert therapeutic effects against alcoholic liver disease and may serve as promising candidates for the development of functional foods beneficial to individuals with excessive alcohol consumption.