基于生物分子通路关联网络的白头翁汤生物效应机制研究

Baitouweng Tang is famous prescription for veterinary clinical treatment diarrhea due to damp-heat, widely used, but its mechanism for the treatment of diarrhea due to damp-heat syndrome is not clear. Referenced the idea of the project network pharmacology, Baitouweng Tang as object, the intervention on diarrhea due to damp-heat syndrome animal model, the use of metabonomics research methods, the ultrahigh pressure liquid chromatography - time of flight mass spectrometry (UPLC/Q - TOF - MS) and temperament - combination (GC - MS) detection of urine, blood plasma, tissue metabolites, the change of the whole, establish metabolic fingerprint screening of Baitouweng Tang and intervention metabolic marker of diarrhea due to damp-heat syndrome; Using KEGG database construction of biological pathways associated metabolic markers of network. On this basis, the intervention of Baitouweng Tang with Cytospace software metabolite markers of diarrhea due to damp-heat syndrome, the channel, the network targets for molecular docking, establish Baitouweng Tang metabolite pathways - target network, this paper Baitouweng Tang intervention diarrhea due to damp-heat syndrome of molecular interaction network, from "point - line - face" to establish a multi-dimensional the interaction relationship between the body and blood tonic soup, to systematically and comprehensively explain the mechanism of action, clinical application and Baitouwang Tang in veterinary medicine provides the basis for new drug research, etc.

白头翁汤为中兽医临床治疗湿热泄泻经典名方，但对其生物效应机制尚不完全清楚。本项目拟以白头翁汤对湿热泄泻模型进行干预，运用代谢组学方法，采用超高压液相色谱-飞行时间质谱法(UPLC/Q-TOF-MS)和气质-联用(GC-MS)分析血浆、尿液、结肠及回肠的代谢物整体变化情况，研究白头翁汤干预湿热泄泻模型的代谢指纹图谱，筛选白头翁汤干预湿热泄泻的纠偏代谢物；利用京都基因与基因组百科全书(KEGG)数据库构建纠偏代谢物的生物通路关联网络。在此基础上，采用网络分析和编辑软件Cytoscape 对白头翁汤干预湿热泄泻的纠偏代谢物、通路、网络靶标进行分子对接，建立白头翁汤代谢物-通路-靶点网络，阐述白头翁汤干预湿热泄泻的生物分子网络交互作用，从“点-线-面”多维度分析白头翁汤与机体之间的相互作用关系，系统地、全面地揭示白头翁汤治疗湿热泄泻的生物效应机制，为湿热泄泻的治疗和中兽药的研究及临床应用提供依据。

白头翁汤为中兽医临床治疗湿热泄泻经典名方，但对其生物效应机制尚不完全清楚。本项目首次在国内开展了基于代谢组学和网络药理学白头翁汤治疗湿热泄泻的生物效应机制研究。研究结果表明，白头翁汤中主要化学成分秦皮甲素、秦皮乙素、药根碱、巴马汀、盐酸小檗碱含量分别为0.7%、0.36%、0.12%、0.38%、8.26%和4.71%。白头翁汤治疗湿热泄泻可能是通过抗炎、增强免疫细胞功能及缓解和保护肠道组织损伤发挥药效作用；湿热泄泻机体的发病机理是5-羟基-N-甲酰犬尿氨酸、左旋谷酰基-左旋半胱氨酸、左旋甲酰犬尿氨酸、亚油酸、褪黑素、核黄素-5-磷酸和左旋色氨酸等潜在代谢标志物含量变化，影响核黄素代谢、色氨酸代谢和亚油酸代谢通路，造成机体代谢系统紊乱；白头翁汤治疗湿热泄泻的作用机理是通过调节湿热泄泻机体内血清生物标志物7-酮基脱氧胆酸、五羟色胺和创伤酸的含量，调控血清甘油磷脂代谢、甘油酯代谢、初级胆酸合成及色氨酸代谢通路；白头翁汤可以调节湿热泄泻机体内尿液潜在代谢标志物的含量，调控谷胱甘肽代谢、色氨酸代谢、磷酸戊糖途径及生物素代谢通路发挥药效作用；白头翁汤可以通过槲皮素、β-谷固醇和豆甾醇等活性成分作用于关键靶点TP53、JUN、IL6、TNF、FOS、IFNG、IL-1β、IL-1α、IL-2和MAPK1，调控炎症性肠病通路发挥药效治疗湿热泄泻。研究建立了白头翁汤代谢物-通路-靶点网络，阐明了白头翁汤干预湿热泄泻的生物分子网络交互作用，为湿热泄泻的治疗和中兽药的研究及临床应用提供依据。在项目资助下发表论文9篇，其中SCI论文4篇，中文核心论文5篇；国家实用新型专利1个；培养博士研究生1名，硕士研究生1名。