Exploring the potential mechanisms of Wumei pills for the treatment of nonalcoholic fatty liver disease based on network pharmacology, molecular docking technology and experimental validation

Objective To investigate the potential mechanism of Wumei Pill (WMP) in improving non-alcoholic fatty liver disease (NAFLD) by regulating lipid metabolism and inflammation via the NF-κB pathway, utilizing network pharmacology, molecular docking, and experimental validation.Methods Active components of WMP and their targets were systematically retrieved from databases including TCMSP. NAFLD-related targets were obtained from disease databases (GeneCards, OMIM). A "drug-component-target" interaction network was constructed using Cytoscape 3.10.1. A protein-protein interaction (PPI) network was built using the STRING platform. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using Metascape. Molecular docking with Autodock Vina 1.5.7 assessed the binding affinity between core components and potential targets. For experimental validation, a NAFLD model was established in C57BL/6 mice fed a methionine-choline deficient (MCD) high-fat diet. Twenty-four mice were randomly divided into model, control, WMP, and fenofibrate groups. After successful modeling, the WMP group received concentrated solution (1.575 g crude drug/mL) via gavage, the fenofibrate group received a 10 mg/mL suspension, while the model and control groups received equivalent saline. After 3 weeks of treatment, serum and liver tissues were collected. Hepatic pathology was assessed by hematoxylin-eosin (HE) and Oil Red O staining. Serum inflammatory markers (TNF-α, IL-6) were measured by ELISA. Serum lipid profiles and liver function markers (ALT, AST, TG, TP, LDL, HDL) were analyzed biochemically. Protein and mRNA expression levels of PPAR-α, PPAR-γ, RAGE, and IGF-1 were analyzed by Western blot and RT-qPCR.Results A total of 137 overlapping targets for WMP in NAFLD treatment were identified. Potential core targets including RELA, TP53, EGFR, AKT1, and RB1 were screened. GO and KEGG enrichment analyses highlighted pathways involved in lipid metabolism, inflammatory response, and oxidative stress, particularly the AGE-RAGE signaling pathways. Molecular docking indicated stable binding between active components and potential targets. Animal experiments demonstrated significantly elevated AST, ALT, and LDL levels in the model group compared to the control group (P < 0.05, P < 0.01, P < 0.001). Compared to the model group, the WMP group significantly reduced levels of TNF-α, IL-6, TG, LDL, and TP (P < 0.05, P < 0.01, P < 0.001), significantly increased IGF-1 protein expression (P < 0.05), and significantly decreased RAGE protein expression (P < 0.05). Pathological examination revealed substantial improvement in liver injury in the WMP group, confirming its therapeutic efficacy against NAFLD.Conclusion Wumei Pill likely ameliorates NAFLD-associated liver injury by inhibiting RAGE protein expression, elevating IGF-1 protein expression, and exerting a multi-component synergistic effect to suppress the RAGE/IGF-1/NF-κB pathway, thereby reducing inflammatory cytokines and lipid accumulation.