Investigating Puerarin's Role in Counteracting Non-Alcoholic Fatty Liver Disease: A Synergistic Study of Network Pharmacology and Experimental Analysis

Objective This study aims to explore the potential molecular mechanisms of puerarin in alleviating non-alcoholic fatty liver disease (NAFLD) by combining network pharmacology and experimental analysis methods. Methods The study identified the active chemical components of puerarin and its potential targets by integrating databases, followed by constructing a target gene database for NAFLD. Through the analysis of the protein-protein interaction (PPI) network and identification of core genes, the focus was on the inhibitory effects of puerarin on inflammatory mediators such as TNF, IL6, IL1B, and STAT3. Laboratory experiments using the HepG2 cell line further validated the hepatoprotective effects of puerarin, demonstrated by reduced levels of ALT and AST, as well as the regulation of stress responses. Quantitative polymerase chain reaction (qPCR) experiments supported these findings, confirming the impact of puerarin on the expression of key inflammatory genes. Results The active components of puerarin and corresponding targets were identified through ETCM2.0 and HERB databases, resulting in the identification of 189 target genes. A total of 2317 target genes related to NAFLD were integrated from GeneCards, DisGeNET, and OMIM databases. Using Venn diagram tools, 188 intersecting target genes between puerarin and NAFLD were determined, and a PPI network was constructed. Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that NAFLD is closely related to biological processes such as insulin resistance, inflammatory response, lipid metabolism disorders, and apoptosis. Experimental validation results showed that puerarin treatment significantly reduced ALT and AST levels in the cell culture supernatant (by 47% and 29%, respectively), decreased MDA levels by 71%, and increased SOD levels by 23%. qPCR experiments indicated that the relative expression levels of IL6, TNF, IL1B, and STAT3 genes in the PA + puerarin treatment group were significantly lower than those in the PA-only treatment group, reduced by 2.0-fold, 2.1-fold, 2.9-fold, and 2.5-fold, respectively. Conclusion This study suggests that puerarin may alleviate the progression of NAFLD by targeting inflammatory pathways and regulating oxidative stress. The combined results of network pharmacology analysis and experimental validation indicate that puerarin modulates the inflammatory response by inhibiting the expression of genes such as IL6, TNF, IL1B, and STAT3, thereby reducing the production and release of inflammatory factors, alleviating liver inflammation, reducing liver fat accumulation, and slowing the progression of NAFLD. Our research provides fundamental data for puerarin as a potential drug for the treatment of NAFLD. With further research and development, puerarin can be designed as an oral medication or other forms of administration or combined with other NAFLD treatments to improve liver health in patients.