Abstract

Background Tinnitus is a common and often debilitating neurological disorder. Phillyrin, a bioactive lignin, has garnered increasing attention for its diverse pharmacological properties. This study utilized an integrated computational approach to investigate the molecular basis underlying phillyrin’s otoprotective effects. Methods A network pharmacology analysis was used to reveal key molecular targets of phillyrin in tinnitus treatment. Protein-protein interaction (PPI) and a “phillyrin-tinnitus-target” interaction networks were generated. GO and KEGG enrichment analyses were performed. Molecular docking was conducted, and conformational dynamics were assessed by normal mode analysis (NMA). In vitro validation was assessed by detecting the viability, apoptosis, and crucial factor expression of mouse spiral ganglion neurons (SGNs) under cisplatin (Cis) treatment. Results This study identified 106 potential targets of phillyrin, 304 tinnitus-related candidate targets, and 11 shared candidates from both aspects. Three proteins (EGFR, HIF1A, and AKT1) were identified as core regulatory nodes. These overlapping targets demonstrated significant enrichment in multiple crucial pathways, including EGFR tyrosine kinase inhibitor resistance, HIF-1, and PI3K-AKT signaling pathways. Molecular docking simulations indicated moderate binding affinity of phillyrin with EGFR (-9.0 kcal/mol), HIF1A (-6.6 kcal/mol), and AKT1 (-6.1 kcal/mol). Conformational dynamics suggested the formation of stable phillyrin-protein complexes with distinct dynamic profiles. Furthermore, phillyrin alleviated Cis-induced cytotoxicity in SGNs and restored EGFR, AKT1, and HIF1A expression. Conclusion This study provides multi-faceted evidence supporting phillyrin as a multi-target therapeutic candidate capable of modulating the EGFR-HIF-1-PI3K-AKT pathways in Cis-induced ototoxicity model, offering novel