Objective: This study aimed to systematically elucidate, for the first time, the therapeutic mechanism of Cuochuangling pills, (CCLP), a clinical herbal formulation for rosacea, through an innovative integrated approach combining network pharmacology, multi-method bioinformatics, and experimental validation. Methods: Active components of CCLP were identified from the TCMSP database. Core therapeutic targets were screened by combining rosacea transcriptome data from GEO with differential expression analysis, weighted gene co-expression network analysis (WGCNA), and three machine learning algorithms. Molecular docking and dynamics simulations were employed to evaluate ligand-receptor binding stability. An LL-37-induced murine rosacea model was established and treated with low-, medium-, and high-dose CCLP for 7 days. Erythema area and severity scores were recorded. Histopathological examination and mast cell infiltration were assessed via H&E and toluidine blue staining, respectively. Inflammatory cytokines (IL-1β, IL-6, TNF-α) were measured by ELISA, and protein expression of TLR2, KLK5, and MMP9 was analyzed by immunohistochemistry. Results: IL-1β was identified as the core target, with KEGG enrichment analysis highlighting inflammatory pathways including NF-κB. Molecular docking revealed a strong binding affinity between quercetin and IL-1β (− 7.4 kcal/mol), further confirmed by stable binding in molecular dynamics simulations. In vivo, medium and high doses of CCLP significantly reduced erythema, severity scores, and mast cell infiltration, while downregulating IL-1β, IL-6, TNF-α, TLR2, KLK5, and MMP9 expression. Conclusion: CCLP alleviates LL-37-induced rosacea-like dermatitis by modulating the TLR2/NF-κB signaling pathway, leading to reduced inflammatory response and mast cell infiltration. Keywords: rosacea, traditional Chinese medicine, network pharmacology, bioinformatics, molecular docking, molecular dynamics simulations
Shen et al. (2026) studied this question.