Unraveling the Mechanism of Cuochuangling Pills in Rosacea Treatment: An Integrated Approach Combining Network Pharmacology, Bioinformatics, and Experimental Validation.

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.