Exploring the mechanism of shikonin in ameliorating allergic rhinitis through integrated bioinformatics analysis, network pharmacology, and experimental validation

Allergic rhinitis (AR) is a nasal mucosal inflammation triggered by allergies. Shikonin, a natural naphthoquinone, has various pharmacological effects. This study sought to clarify how shikonin alleviates AR. The identification of differentially expressed genes (DEGs) in AR and GO/KEGG analyses of related genes was performed using SangerBox. Targets of shikonin were collected using the Traditional Chinese Medicine Systems Pharmacology Database, Similarity Ensemble Approach database, and SymMap, while AR-related targets were gathered from GeneCards and the Therapeutic Target Database. Protein-protein interaction networks for shikonin-AR shared targets, as well as pathway-target-immune correlation analyses, were conducted using the search tool for the retrieval of interacting genes/proteins. Molecular docking of shikonin with tumor necrosis factor (TNF) pathway proteins was performed using AutoDock Vina. Cell viability was assessed via the Cell Counting Kit-8 assay, and protein expression was detected by western blot. DEGs in AR were significantly enriched in the TNF pathway, and the TNF pathway played a crucial role in the interaction between shikonin and AR. Molecular docking revealed that shikonin formed stable binding with TNF pathway proteins (TNF, TNFAIP3, and NFKB1). Furthermore, shikonin impacted the TNF pathway via a multi-target, multi-pathway mechanism, and there was a profound connection between this pathway and immune regulation. Additionally, shikonin was capable of mitigating the damage induced by IL-13 on nasal epithelial cells by inhibiting the activation of the TNF pathway. Shikonin may alleviate the symptoms of AR through the TNF pathway, providing insights for understanding the pathogenesis of AR and the exploration of novel therapeutic strategies.