Synergistic Effect of Combined Wheat Peptides, Soft‐Shelled Turtle Peptides and <i>Pueraria Lobata</i> Root Extract on Alleviating Alcoholic Liver Disease by Regulating Gut Microbiota and Intestinal Barrier

This study aimed to investigate the hepatoprotective effects and underlying mechanisms of a peptide-plant extract combination against alcohol-induced liver injury in a zebrafish model. The results demonstrate that wheat peptides (WP), soft-shelled turtle peptides (STP), and Pueraria lobata root extract (PE) significantly attenuate hepatomegaly and lipid droplet accumulation, enhance the activities of acetaldehyde dehydrogenase (ALDH) and superoxide dismutase (SOD), and decrease the levels of aspartate aminotransferase (AST) and malonaldehyde (MDA). The optimal formulation ratio of WP, STP, and PE (1:4:1) is determined using fuzzy comprehensive evaluation (FSE). The combination significantly down-regulated the expression of the alcohol-metabolism-related genes cytochrome P450 2Y3, cytochrome P450 (CYP)(cyp2y3) and cytochrome P450CYP3A65 (cyp3a65), the fatty acid synthesis-related gene acetyl-CoA carboxylase 1 (acc1), and DNA damage-inducing gene 153 (GADDl53) (chop). Furthermore, the combination improves gut microbiota diversity, increases the relative abundance of Chitinilyticum, and decreases that of ZOR006 and Erysipelotrichaceae. Non-targeted metabolomics analysis reveals that the hepatoprotective effect is primarily mediated through by up-regulating of nucleotide and pyrimidine metabolism pathways, thereby maintaining cellular energy homeostasis. In addition, the combination up-regulates ZO-1 and Nrf-2 expression, contributing to the restoration of alcohol-induced intestinal barrier integrity. Collectively, the optimized peptide-plant extract formulation confers significant protection against alcohol-induced liver injury by modulating gut microbiota composition, re-establishing hepatic metabolic homeostasis, and reinforcing intestinal barrier function.