RESUMEN
We explored the mechanism of patchouli oil in the treatment of inflammatory bowel disease (IBD) based on network pharmacology and differentially expressed genes in macrophages. The chemical composition of patchouli oil was detected by GC-MS, targets for active components were collected through TCMSP and Swiss Target Prediction platform, and targets for treatment of IBD were retrieved from DrugBank, GeneCards, OMIM, PharmGkb, and TTD databases. The intersection targets were merged, Cytoscape software was used to construct the "component-to-intersection target" network, and protein-protein interaction (PPI) network was drawn with String platform. The intersection targets were enriched for GO and KEGG enrichment analysis on Metascape platform, and the molecular docking of AutoDock Vina was used to verify the analysis results. The macrophage chip data was downloaded, and the differential genes were obtained by using R software. KEGG signaling pathway analysis of differentially expressed genes were performed by DAVID platform. Real-time fluorescence quantitative PCR was used to verify the screened components in the cell model in vitro. The 14 main components of patchouli oil corresponded to 112 targets, and the intersection obtained 97 common targets of patchouli oil for IBD treatment. GO enrichment analysis yielded 53 items. Eighteen items were obtained by KEGG enrichment analysis, involving cAMP signaling pathway, Notch signaling pathway, adhesion connection, Th17 cell differentiation and other signaling pathways. Molecular docking showed that the selected active components of patchouli oil had good binding activity with the targets. Differentially expressed genes were enriched in inflammatory pathways such as Toll-like receptors, JAK-STAT and NF-κB signaling pathways. q-PCR showed that patchouli oil, patchouli alcohol, pogostone can reduce the mRNA levels of cytokines (TNF-α, IL-1β, IL-6, and IL-23) and up-regulate the mRNA levels of tight junction proteins (occludin and claudin-1) in the inflammatory model of NCM460 normal colon epithelial cells. Patchouli alcohol can significantly reduce the levels of TNF-α, IL-6, and IL-1β inflammatory factors in RAW264.7 macrophages induced by LPS. This study revealed the multi-component, multi-target and multi-pathway of patchouli oil, and confirms the anti-inflammatory effect of patchouli oil and its main components in the inflammatory cell model in vitro and the protection of intestinal epithelial barrier integrity function, which provides a theoretical basis for further elucidating the mechanism of patchouli oil in the treatment of IBD.