Integrative Network Pharmacology, Molecular Docking, and In Vitro Insights Into the Mechanism of Finger Citron for...

Finger citron (FC), a traditional plant with both pharmacological activity and food-grade safety, is considered to possess potential for the improvement of non-alcoholic fatty liver disease (NAFLD). However, the bioactive constituents responsible for this effect and their underlying mechanisms remain incompletely understood. In this study, an integrative approach combining network pharmacology, molecular docking, and in vitro validation was employed to elucidate the mechanisms by which FC exerts anti-NAFLD effects. Network pharmacology analysis identified 62 bioactive compounds targeting 306 liver injury-related genes, among these, 3,4,7-trimethoxycoumarin (TMC) was selected for further investigation. Gene Ontology (GO) enrichment revealed that FC-mediated effects were significantly associated with protein phosphorylation, inflammatory responses, and protein kinase activity, whereas Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment highlighted key involvement of the PI3K-Akt and MAPK signaling pathways. Molecular docking demonstrated high binding affinities between TMC and key target proteins including IL-6, TNF-α, albumin (ALB), AKT1, and STAT3. The successful synthesis of TMC was confirmed via ¹H-NMR and MS analyses. In vitro studies revealed that TMC effectively reduced lipid accumulation and oxidative stress induced by free fatty acids in HepG2 cells. Furthermore, RT-qPCR analysis showed that TMC modulated the mRNA expression of key target genes and lipid metabolism-associated genes, including CPT2, and APOC2, thereby substantiating the mechanistic basis of its therapeutic action. Collectively, these findings highlight the therapeutic potential of FC against NAFLD and provide a scientific basis for its further development as a promising NAFLD intervention.