基于结肠代谢组学和网络药理学整合策略探究参苓白术散治疗溃疡性结肠炎的作用机制

Integrated strategy for mechanism of Shenling Baizhu San in treating ulcerative colitis based on colonic metabolomics and network pharmacology

参苓白术散是治疗溃疡性结肠炎(ulcerative colitis,UC)的常用复方,该研究采用结肠代谢组学整合网络药理学探究其治疗UC的作用机制。BALB/c小鼠随机分成空白组、模型组、参苓白术散组、柳氮磺胺吡啶组,运用UPLC-Q-TOF-MS/MS技术对小鼠结肠组织代谢谱进行分析,筛选差异代谢物及相关代谢通路;基于在线数据库筛选参苓白术散活性成分及作用靶点、UC疾病靶点,利用STRING和Cytoscape 3.9.1构建参苓白术散治疗UC核心靶点的蛋白-蛋白互作网络,采用DAVID数据库进行基因本体(GO)功能和京都基因组百科全书(KEGG)通路富集,并构建“代谢物-反应-酶-基因”网络进行代谢组学与网络药理学联合分析。参苓白术散对UC结肠组织中参与D-谷氨酰胺和D-谷氨酸代谢,咖啡因代谢,鞘脂类代谢,精氨酸生物合成,赖氨酸降解,丙氨酸、天冬氨酸和谷氨酸代谢,甘油磷脂代谢和嘧啶代谢等代谢通路的25种代谢物有显著回调趋势;参苓白术散治疗UC的47个核心靶点参与了MAPK信号通路、TNF信号通路、Toll样受体信号通路、脂质和动脉粥样硬化、炎症性肠病、Th17细胞分化等。整合分析显示甘油磷脂代谢和嘧啶代谢是参苓白术散治疗UC的关键代谢途径。结果表明参苓白术散可以通过调节结肠代谢物改善结肠黏膜形态,调控炎症相关核心靶基因降低炎症水平,缓解机体脂质代谢紊乱发挥治疗UC的作用。

Shenling Baizhu San(SLBZS)is a commonly used medicine for the treatment of ulcerative colitis(UC).This study aims to explore the mechanism of SLBZS in treating UC by using colonic metabolomics and network pharmacology.BALB/c mice were randomly divided into four groups:a blank group,a model group,an SLBZS group,and a sulfasalazine group.UPLC-Q-TOF-MS/MS technology was utilized to analyze the metabolic profiles of colonic tissue in mice,and differential metabolites and related metabolic pathways were screened.Based on the online database,active ingredients,action targets,and UC disease targets of SLBZS were screened.The protein-protein interaction(PPI)network of core targets of SLBZS in treating UC was constructed using STRING and Cytoscape 3.9.1.Gene Ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed using the DAVID database.A"metabolite-reaction-enzyme-gene"network was constructed to conduct a combined analysis of metabolomics and network pharmacology.SLBZS reversed the levels of 25 metabolites involved in various pathways such as D-glutamine and D-glutamate metabolism,caffeine metabolism,sphingolipid metabolism,arginine biosynthesis,lysine degradation,alanine,aspartate,and glutamate metabolism,glycerophospholipid metabolism,and pyrimidine metabolism in UC colonic tissue.47 core targets of SLBZS in treating UC were involved in pathways including the MAPK signaling pathway,TNF signaling pathway,Tolllike receptor signaling pathway,lipid and atherosclerosis,inflammatory bowel disease,and Th17 cell differentiation.Integrated analysis showed that glycerophospholipid metabolism and pyrimidine metabolism were key metabolic pathways in the treatment of UC with SLBZS.The results suggested that SLBZS improved colonic mucosal morphology by regulating colonic metabolites,down-regulated the expression of inflammation-related core target genes to reduce inflammation levels,and alleviated lipid metabolism disorders,thereby exerting a therapeutic effect on UC.