Network pharmacology combined with molecular docking and experimental validation reveals the anti-pharyngitis...

This study explored the molecular mechanisms of Humei Qingyan granule (HMQY) in treating pharyngitis via network pharmacology, molecular docking, and in vivo experiments. Public databases were used to collect HMQY's main ingredients, their targets, and pharyngitis-related targets. The protein-protein interaction (PPI) network was constructed using the STRING 11.0 database, with hub targets pinpointed via Cytoscape. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were employed to predict biological processes and pathway, followed by molecular docking of key components and targets. Finally, an acute pharyngitis rat model was used to validate the predicted targets and pathways, through enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR), and western blot assays. Results revealed active compounds like kaempferol, quercetin, and physovenine, with hub targets including AKT1, TP53, VEGFA, CASP3, and IL-1β. KEGG analysis highlighted the PI3K-Akt pathway and inflammation-cancer crosstalk as critical. Molecular docking showed strong binding between AKT1 and HMQY's core components. In vivo experiments confirmed HMQY effectively alleviated pharyngeal pathological damage in rats with acute pharyngitis, and molecular biology experiments further revealed that HMQY treated pharyngitis by regulating the PI3K-Akt pathway, as shown by the inhibition of the protein expression of PI3K, p-Akt, and p-NF-κB, and a significant reduction in the level of IL-1β. In conclusion, this study preliminarily validated the active ingredients, key targets, and potential pathways of HMQY in the treatment of pharyngitis. The findings suggest HMQY exerts its therapeutic effects on pharyngitis primarily by inhibiting the PI3K-Akt/NF-κB signaling pathway.