Immunomodulatory Mechanism of Baiyaojian Decoction on Periodontitis: Network Pharmacology, Single-Cell RNA Sequencing and Molecular Docking

白药健汤治疗牙周炎的免疫调节机制:网络药理学、单细胞RNA测序和分子对接

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Abstract

Periodontitis is one of the most common oral inflammatory diseases. Baiyaojian decoction, known for its prominent immunomodulatory and anti-inflammatory properties, shows significant potential in treating periodontitis, though its molecular mechanisms remain unknown. The active ingredients and therapeutic targets were determined by integrating multiple databases. The protein-protein interaction network was constructed by the STRING platform. Bulk RNA seq data of GSE16134 were included and GO enrichment, GSEA and CIBERSORT algorithm were employed to investigate the immune microenvironment in periodontitis. Single-cell RNA seq data of GSE152042 and GSE171213 were integrated by harmony; the cell-cell communication network was analysed by CellChat, and the differentiation trajectory was constructed by monocle3. Molecular docking was carried out using AutoDockTools, AutoDock Vina and PyMOL. Experimental validation was performed via qRT-PCR, CCK-8 assay, flow cytometry and ELISA. Twenty-seven active ingredients and 207 therapeutic targets were obtained. Thirty-one core therapeutic targets were identified. The infiltration of plasma cells, neutrophils, macrophages and mast cells was significantly enhanced in periodontitis tissues. Twenty-eight of 31 core therapeutic targets were involved in their infiltration, differentiation and pro-inflammatory activities. Molecular docking suggested stable bindings between ingredients and therapeutic targets. Experimental validation confirmed the elevated infiltration of above immune cells and demonstrated the anti-inflammatory properties and target modulation capabilities of key ingredients including Coumestrol, Diosmetin and Gallicin. Baiyaojian decoction may exert immunomodulatory and anti-inflammatory effects to treat periodontitis through multi-ingredient and multi-target mechanisms.

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