Mechanism of Artemisia argyi Levl.et Vant. aqueous extract in alleviating dextran sulfate sodium-induced ulcerative colitis

艾蒿水提取物缓解葡聚糖硫酸钠诱导的溃疡性结肠炎的机制

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Abstract

This study aims to explore the effect of Artemisia argyi Levl.et Vant. (AAL) on the intestinal microbiota in UC mice and the possible mechanism of AAL in alleviating UC, providing new scientific evidence for clinical application. The chemical ingredients of AAL aqueous extract were analyzed using ultra-performance liquid chromatography Q-exactive mass spectrometry (UHPLC-QE-MS) non-target metabolomics. UC model mice were induced by 3% dextran sulfate sodium (DSS) to observe the ameliorative effect of AAL on colonic inflammation in UC mice. Mouse intestinal microbial diversity, community composition, and community function were analyzed using 16 S rRNA gene sequencing, linear discriminant analysis effect size (LEfSe), and phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt). The key targets of AAL for UC treatment were clarified using network pharmacology. The binding activities of AAL characteristic ingredients and core targets were matched by molecular docking technology. Finally, the mechanism of AAL treatment for UC was confirmed by immunohistochemistry (IHC) assay. UHPLC-QE-MS non-target metabolomics showed that the active ingredients of AAL aqueous extract with high stability and reliability can be detected in both positive and negative ion modes. AAL can alleviate colonic inflammation and reduce pathological damage to colonic tissues in UC mice. Intestinal microbial community analysis demonstrated that AAL can restore intestinal flora imbalance in UC mice by increasing the abundance of beneficial bacteria and decreasing the abundance of harmful bacteria. Network pharmacology revealed that TNF-α, IL-6, IL-1β, PTGS2, and ESR1 are key targets of AAL for UC treatment. The molecular docking results showed that 1,8-cineole and borneol formed stable complexes with all five core targets, and the complexes had high binding affinity. IHC assays showed that AAL inhibited the activation of IL-17 signaling pathway and regulated the ferroptosis pathway by down-regulating IL-17 A, IL-17RA, Act1, TRAF6, and PTGS2 expression and up-regulating GPX4 expression. AAL alleviates DSS-induced UC by synergistically inhibiting the inflammatory response, regulating intestinal flora dysbiosis, modulating the IL-17 signaling pathway, and suppressing the ferroptosis pathway. These findings preliminarily elucidate the mechanism of AAL in alleviating UC and potentially provide a theoretical basis for the clinical treatment of UC and the development of new drugs.

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