摘要
目的运用UHPLC-QTOF-MS和网络药理学探讨温阳化浊退黄方治疗慢加急性肝衰竭(Acute-on-chronic liver failure,ACLF)的作用机制,并通过实验验证筛选出的靶点。方法利用UHPLCQTOF-MS分析温阳化浊退黄方的活性成分,利用网络药理学方法筛选温阳化浊退黄方活性成分对应的靶点和ACLF靶点,构建蛋白互作网络(PPI)筛选核心靶点,对靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)通路分析,利用Cytoscape构建活性成分-靶点-通路网络图,通过动物模型实验验证温阳化浊退黄方治疗ACLF的作用机制。结果UHPLC-QTOF-MS共鉴定出123个化合物,经TCMSP筛选得到12种活性成分,TCMSP和Swiss target Prediction数据库取交集共获得181个温阳化浊退黄方预测靶点;得到ACLF靶点2019个;温阳化浊退黄方和ACLF共有靶点99个。KEGG通路分析筛选了147条相关信号通路,显示PI3K/Akt信号通路、VEGF信号通路、乙型肝炎、细胞凋亡、MAPK信号通路等可能在温阳化浊退黄方治疗ACLF的过程中起关键作用。动物实验结果显示,温阳化浊退黄方能够缓解ACLF大鼠肝损伤,下调AKT1、MTOR和RAF1关键靶蛋白在ACLF大鼠肝脏中的表达,验证了网络药理学的预测结果。结论温阳化浊退黄方通过多成分、多靶点、多通路协同作用治疗ACLF,为后续研究提供参考依据。
Objective To explore the mechanism of Wenyang Huazhuo Tuihuang Fang in the treatment of acute on-chronic liver failure(ACLF)by UHPLC-QTOF-MS and network pharmacology,and to verify the screened target by experiments.Methods UHPLC-QTOF-MS was used to analyze the active components of Wenyang Huazhuo Tuihuang Fang,and network pharmacology was used to screen the targets corresponding to the active components of Wenyang Huazhuo Tuihuang Fang and ACLF targets,and protein interaction network(PPI)was constructed to screen the core targets.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed on the targets.Cytoscape was used to construct the active ingredient-target-pathway network diagram,and animal model experiments were performed to verify the mechanism of action of Wenyang Huazhuo Tuihuang Fang in the treatment of ACLF.Results A total of 123 compounds were identified by UHPLC-QTOF-MS,and 12 active components were screened by TCMSP.A total of 181 Prediction targets were obtained by intersection of TCMSP and Swiss Target Prediction database.A total of 2019 targets of ACLF were obtained.There were 99 targets of Wenyang Huazhuo Tuihuang Fang and ACLF.KEGG pathway analysis screened 147 related signaling pathways,indicating that PI3K/Akt signaling pathway,VEGF signaling pathway,hepatitis B,apoptosis,MAPK signaling pathway may play a key role in the treatment of ACLF by Wenyang Huazhuo Tuihuang Fang.Animal experimental results showed that Wenyang Huazhuo Tuihuang Fang could alleviate liver injury in ACLF rats and down-regulate the expression of AKT1,MTOR and RAF1 key target proteins in the liver of ACLF rats,confirming the prediction results of network pharmacology.Conclusion Wenyang Huazhuo Tuihuang Fang can treat ACLF through multi-component,multi-target and multi-pathway synergistic action,which provides reference for subsequent studies.
作者
王秀峰
张荣臻
陈月桥
王明刚
吕超
Wang Xiufeng;Zhang Rongzhen;Chen Yueqiao;Wang Minggang;Lyu Chao(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第5期1859-1870,共12页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
广西壮族自治区科学技术厅青年科学基金项目(2018GXNSFBA281031):miRNA调控慢加急性肝衰竭大鼠Th17/Treg平衡及温阳化浊退黄方的干预研究,负责人:王秀峰
广西壮族自治区科学技术厅创新研究团队项目(2018GXNSFGA281002):肝衰竭发病机制及中医药防治,负责人:毛德文
