Ulcerative colitis (UC) is a persistent, diffuse intestinal inflammation and ranks among the most challenging chronic diseases worldwide. Atractylodes lancea (Thunb.) DC. and Atractylodis macrocephala Koidz. are traditional Chinese medicines (TCMs) with a long history of clinical application, particularly for gastrointestinal disorders. Both Atractylodis Rhizoma (AR) and Atractylodis Macrocephala Rhizoma (AM) have shown significant efficacy in managing UC; however, the underlying mechanism by which the AR-AM herbal pair promotes intestinal mucosal healing remains poorly understood. The therapeutic effects of the ethanolic extract of AR-AM (EEAR-AM) were evaluated in a murine UC model induced by dextran sodium sulfate (DSS). A network pharmacology approach was employed to explore the anti-UC properties of EEAR-AM, including identification of active compounds, prediction of potential targets, and construction of a protein-protein interaction (PPI) network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were subsequently performed to preliminarily elucidate the mechanisms of EEAR-AM in UC treatment. Finally, the proposed molecular mechanisms were validated in both DSS-induced UC mice and Caco-2 cells. In vivo results demonstrated that EEAR-AM significantly attenuated DSS-induced weight loss, reduced colon shortening, lowered the disease activity index (DAI) score, and modulated the spleen coefficient. Moreover, EEAR-AM improved colonic tissue architecture, reduced inflammatory infiltration, restored goblet cell density, enhanced mucin MUC2 expression, and elevated levels of tight junction (TJ) proteins. Additionally, EEAR-AM suppressed the expression of matrix metalloproteinase 2 (MMP-2) and MMP-9. Network pharmacology analyses indicated that EEAR-AM may ameliorate intestinal mucosal dysfunction through modulation of the exchange protein directly activated by cAMP 1 (Epac1)/Ras-associated protein 1 (Rap1) pathway and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathways. These actions potentially enhance cellular barrier integrity and reduce the release of inflammatory mediators. Western blotting results confirmed that EEAR-AM activated the Epac1/Rap1 pathway while downregulating the PI3K/AKT pathway in both DSS-induced UC mice and Caco-2 cells, consistent with predictions from network pharmacology. This study represents the first evidence that the EEAR-AM herbal pair improves intestinal mucosal barrier function in UC, with therapeutic effects likely mediated by activation of the Epac1/Rap1 pathway and inhibition of the PI3K/AKT pathway.
Funding
This work was supported by the Key Scientific ResearchProject of Hubei Provincial Department of Education (No. D20232001).
Supporting information
Supporting materials of this paper can be requested by sending E-mail to the corresponding authors.
Declaration of competing interest
These authors declare no conflict of interest.
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