基于尿液代谢组学和网络药理学整合策略研究白头翁汤治疗湿热泄泻的作用机制  

Integrated strategy for mechanism of Baitouweng Decoction in treating dampness-heat diarrhea based on urine metabolomics coupled with network pharmacology

作　　者：华永丽[1] 马琪 张晓松[1] 纪鹏[1] 袁子文[1] 魏彦明[1] HUA Yong-li;MA Qi;ZHANG Xiao-song;JI Peng;YUAN Zi-wen;WEI Yan-ming(College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China;College of Animal Science,Southucest University,Chongqing 402460,China)

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]西南大学动物科学学院,重庆402460

出　　处：《中国中药杂志》2022年第14期3887-3897,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(31860717);甘肃省高等学校创新基金项目(2020A-057);甘肃农业大学伏羲杰出人才培育计划项目(Gaufx-03J01);财政部和农业农村部国家现代农业产业技术体系项目(CARS-37)。

摘　　要：白头翁汤为临床治疗湿热泄泻(dampness-heat diarrhea)的经典方剂,但其治疗湿热泄泻的作用机制不明确。该研究采用尿液代谢组学整合网络药理学的方法研究其治疗湿热泄泻的作用机制。Wistar大鼠随机分为对照组、白头翁汤组(3.6 g·kg)、湿热泄泻组和自愈组;采用超高效液相色谱四极杆飞行时间质谱(UPLC-Q-TOF-MS)联用技术进行各组尿液代谢谱分析,多变量统计分析筛选差异代谢物;基于中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform, TCMSP)筛选白头翁汤活性成分及其靶标,基于在线数据库筛选腹泻相关蛋白靶标。使用Cytoscape 3.2.1建立白头翁汤化学成分-靶标蛋白质相互作用网络,并建立白头翁汤活性成分作用靶标、腹泻相关蛋白和差异代谢产物相关蛋白的蛋白-蛋白相互作用(PPI)网络,并将3个PPI网络合并。基于Cytoscape 3.2.1的ClueGO相关遗传相似性进行基因富集分析。与对照组相比,湿热泄泻组中N-乙酰-5-羟色胺、L-γ-谷氨酰半胱氨酸、谷胱甘肽、视黄酸、褪黑素、吲哚-3-乙醛、L-胱氨酸、生物素和L-色氨酸相对含量有明显差异,白头翁汤可以显著回调这些代谢产物的相对含量。基因富集分析表明167个靶标与活性氧的产生、炎症发生和细胞凋亡有关。结果表明,白头翁汤可以通过抑制炎症发生及活性氧产生,阻断细胞凋亡发挥治疗湿热泄泻的作用。Baitouweng Decoction is a famous Chinese medicinal decoction that has been used to treat diarrhea over thousands of years. In this study, we investigated the effect and mechanism of Baitouweng Decoction in the treatment of diarrhea. Wistar rats were randomly assigned into 4 groups: control group, dampness-heat diarrhea model group(modeling by complex factors including high-sugar and high-fat diet, improper diet, hot and humid environment, drinking and intraperitoneal injection of Escherichia coli), Baitouweng Decoction(3.6 g·kg) group, and self-healing group. A urine metabolomics approach was developed with ultra liquid chromatography-quadrupole-time-of-flight-mass spectrometry(UPLC-Q-TOF-MS) for metabolic profiling. The differential metabolites were screened out by the multivariate comparison between groups. Diarrhea-related protein targets and the active compounds of Baitouweng Decoction were used to predict the protein targets of Baitouweng Decoction. Cytoscape 3.2.1 was employed to establish a active component-target protein interaction network. Three protein-protein interaction(PPI) networks of component target proteins, diarrhea-related proteins, and differential metabolite-related proteins were established and then merged by BisoGenet. ClueGO was used to perform the gene enrichment based on the genetic similarity. The results showed that Baitouweng Decoction effectively treated dampness-heat diarrhea in vivo. N-acetylserotonin, L-gamma-glutamylcysteine, glutathione, retinoate, melatonin, indole-3-acetaldehyde, L-cystine, biotin, and L-tryptophan were screened as differential metabolites in dampness-heat diarrhea model group. Tryptophan metabolism, glutathione metabolism, biotin metabolism, retinol metabolism, and cysteine and methionine metabolism were involved in the therapeutic effect of Baitouweng Decoction in vivo. A total of 167 targets were identified as major candidates for diarrhea progression. The gene-set enrichment revealed that the targets were involved in reactive oxygen species production, i

关 键 词：白头翁汤 代谢组学 网络药理学 湿热泄泻 

分 类 号：R285[医药卫生—中药学]