基于UPLC-MS/MS和网络药理学分析杨树桑黄水提上清物降血糖作用机制

    Study on the Hypoglycemic Effect Mechanismof the Water-extracted Supernatant of Sanghuangporus vaninii by UPLC-MS/MS and Network Pharmacology

    • 摘要: 为研究杨树桑黄水提上清物活性成分及其降血糖作用机制。采用超高效液相色谱-质谱联用法(UPLC-MS/MS)分析杨树桑黄水提上清物活性成分,以明确其化学成分;借助SwissADME和SwissTargetPredition数据库,对已鉴定出的活性成分的潜在靶点进行预测;;同时,利用GeneCards、DrugBank和OMIM数据库获取2型糖尿病相关靶点,将成分靶点与疾病靶点取交集得到潜在靶点。进一步通过Cytoscape 3.10.3软件构建“药物-成分-靶点”网络,STRING 数据库构建 PPI 网络,DAVID 数据库进行基因本体论(GO)功能富集分析和京都基因与基因组百科全书(KEGG)通路富集分析,AutoDockTools 1.5.6 软件完成分子对接,结果通过PyMOL 2.6进行可视化分析。结果表明,杨树桑黄水提上清物经UPLC-MS/MS鉴定,共发现1 453种化合物;通过数据库筛选预测得到507个活性成分和955个SVWS治疗T2DM的潜在靶点。GO富集分析显示,核心靶点主要富集在以下功能和通路,在生物过程包括RNA聚合酶II介导的转录正调控、DNA模板化转录的正调控和基因表达的正调控等;细胞组成包括细胞质溶胶、细胞核和细胞质等;分子功能包括同源蛋白结合、酶结合和DNA结合转录因子活性等。KEGG通路富集分析显示,核心靶点主要富集在乙型肝炎、脂质代谢与动脉粥样硬化、IL-17信号通路、癌症通路和糖尿病并发症中的AGE-RAGE信号通路等信号通路。分子对接结果表明,核心成分与核心靶点可稳定结合,结合能均低于 −5.0 kcal·mol−1。综上,杨树桑黄水提上清物活性成分通过多成分-多靶点-多通路的方式发挥降糖作用,推测SVWS中6个核心活性成分(6,7,4-三羟基黄酮、苦苣内酯、表鲁比敏、橙皮素、短叶松素-乙酸酯和球松素)可能通过11个关键靶蛋白(PPARG、AKT1、GAPDH、PTGS2、TNF、IL6、IL1B、CXCL8、MAPK1、MAPK3、MAPK14),调节乙型肝炎和脂质和动脉粥样硬化信号通路,以此达到降糖作用,为开发具有降糖作用的天然药物提供理论依据。

       

      Abstract: To investigate the bioactive components of the Water-extracted Supernatant of Sanghuangporus vaninii(SVWS)and its hypoglycemic mechanism, an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)approach was employed for component analysis. The potential targets of these bioactive components were predicted using the SwissADME and SwissTargetPrediction databases, while the targets associated with type 2 diabetes mellitus(T2DM)were retrieved from the GeneCards, DrugBank and OMIM databases. The intersection of component targets and disease targets was obtained to identify the potential therapeutic targets of SVWS for T2DM. The "drug-component-target" network was constructed via Cytoscape 3.10.3, the protein-protein interaction(PPI)network was built using the STRING database, and Gene Ontology(GO)functional enrichment analysis as well as Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed on the DAVID database. Molecular docking was accomplished with AutoDockTools 1.5.6, and the results were visualized by PyMOL 2.6.The results demonstrated that a total of 1,453 compounds were identified in the Water-extracted Supernatant of Sanghuangporus vaninii by UPLC-MS/MS. Through database screening and prediction, 507 bioactive components and 955 potential targets of SVWS for T2DM treatment were obtained. GO enrichment analysis revealed that the core targets were mainly enriched in the following biological processes, cellular components and molecular functions: biological processes included positive regulation of transcription by RNA polymerase II, positive regulation of DNA-templated transcription and positive regulation of gene expression; cellular components involved cytosol, nucleus and cytoplasm; molecular functions comprised identical protein binding, enzyme binding and DNA-binding transcription factor activity. KEGG pathway enrichment analysis indicated that the core targets were primarily enriched in Hepatitis B, Lipid and atherosclerosis, IL-17 signaling pathway, Pathways in cancer, and AGE-RAGE signaling pathway in diabetic complications. Molecular docking results showed that the core components could bind stably to the core targets, with all binding affinities lower than -5.0 kcal·mol-1.In conclusion, the bioactive components of the Water-extracted Supernatant of Sanghuangporus vaninii exert hypoglycemic effects through a multi-component, multi-target and multi-pathway mode. It is speculated that six core bioactive components in SVWS(6,7,4-Trihydroxyflavanone, Sonchifolin, Epilubimin, Hesperetin, Pinobanksin acetate, Pinostrobin)may exert hypoglycemic effects by regulating the Hepatitis B and Lipid and atherosclerosis signaling pathways via eleven key target proteins(PPARG, AKT1, GAPDH, PTGS2, TNF, IL6, IL1B, CXCL8, MAPK1, MAPK3 and MAPK14). This study provides a theoretical basis for the development of natural hypoglycemic drugs.

       

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