Integrated network pharmacology, molecular docking analysis and experimental validation to reveal the mechanisms of Jinglian Hexiao formula in the treatment of adenoid hypertrophy

Objective: Jinglian Hexiao formula (JLHXF) has shown promising therapeutic effects on adenoid hypertrophy (AH) in clinical practice, although its underlying mechanism remains unclear. This study aimed to excavate the pharmacological substance basis of JLHXF for the treatment of AH and reveal its mechanism of action. Methods: The pharmacological mechanisms of JLHXF in treating AH were investigated using network pharmacology and molecular docking approaches. In vitro experiments were then conducted to validate the molecular mechanisms. Results: A total of 96 active components and 13,021 potential target genes of JLHXF were screened and 22 key components and 10 core targets were selected through target network analysis. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed a significant association, it was found that the interleukin-4 and interleukin-13 signaling pathway was closely linked to AH. Molecular docking of core protein-ligand interactions was performed using AutoDock software, the molecular docking results showed that quercetin, luteolin, ursolic acid and beta-sitosterol of JLHXF could bind to BCL2, respectively. In vitro experiments showed that JLHXF could significantly decrease the cellular inflammatory response induced by LPS, reduce the secretion levels of IL6 and TNF-α, down-regulate the expression of IL6, TNF-α, INOS and COX-2 mRNA, as well as reduce the expression levels of pro-apoptotic proteins Bax and caspase-12. Conclusion: These findings indicate that JLHXF exerts anti-inflammatory effects by suppressing the expression of inflammatory factors and pathways, concurrently exerting anti-apoptotic effects, thus exerting a therapeutic effect on AH.