基于UPLC/LTQ-Orbitrap-MS和网络药理学的桔梗治疗非酒精性脂肪肝病的机制研究

  目的  结合UPLC/LTQ-Orbitrap-MS技术和网络药理学方法预测桔梗治疗非酒精性脂肪肝病(NAFLD)的作用靶点及潜在的作用机制，并进行相关实验验证，为深入揭示桔梗治疗NAFLD的药效物质及作用机制奠定基础。  方法  通过UPLC/LTQ-Orbitrap-MS技术及中药系统药理学数据库与分析平台(TCMSP)数据库筛选桔梗的活性成分；使用Swiss Target Prediction数据库对桔梗的活性成分进行靶点预测；同时通过OMIM、Disgenet、TTD等数据库获取NAFLD靶点；将疾病相关靶点映射到化合物潜在靶点中，获取公共靶点，并将信息导入Cytoscape软件和String在线分析平台分别制作网络图和PPI图，同时进行拓扑学分析；基于R软件使用Bioconductor生物信息软件包进行关键靶基因GO与KEGG功能富集分析。构建NAFLD小鼠模型，通过病理染色切片，qPCR实验验证网络药理学富集分析结果。  结果  结合质谱分析与数据库筛选结果，共获得桔梗活性成分13个，药物靶点278个，疾病靶点1 536个，共同靶点83个，涉及PI3K-AKT、胰岛素抵抗、TNF-α、IL-17、JAK-STAT、T细胞受体等信号通路。实验验证显示关键靶基因存在差异性表达。  结论  该研究初步揭示了桔梗治疗NAFLD的活性成分及其作用机制，为后续的深入研究提供了参考价值。 

UPLC/LTQ-Orbitrap-MS Combined with Network Pharmacology to Explore the Mechanism of Platycodon Grandiflorum in the Treatment of Nonalcoholic Fatty Liver Disease

OBJECTIVE  Combining UPLC/LTQ-Orbitrap-MS technology with network pharmacology method to predict the target of Platycodon grandiflorum in the treatment of non-alcoholic fatty liver disease (NAFLD) and the potential mechanism, and carrying out relevant experimental verification, in order to reveal the effective substances and mechanism of Platycodon grandiflorum in the treatment of NAFLD.METHODS  The active components of Platycodon grandiflorum were screened by UPLC/LTQ-Orbitrap-MS and TCMSP database. Swiss Target Prediction Database was used to predict the active components of Platycodon grandiflorum. Targets of NAFLD were obtained through OMIM, Disgenet, TTD and other databases. The disease-related targets were mapped to the potential targets of compounds to obtain the common targets, and the information was imported into Cytoscape software and String online analysis platform to make network diagram and PPI diagram, respectively, and topological analysis was carried out at the same time. Functional enrichment analysis of key target genes GO and KEGG was carried out by using Bioconductor bioinformatics software package based on R software. NAFLD mouse model was established, the results of network pharmacology enrichment analysis were verified by pathological staining and qPCR.RESULTS  Combined with the results of mass spectrometry and database screening, a total of 13 active components, 278 drug targets, 1 536 disease targets and 83 common targets were screened in Platycodon, involving PI3K-AKT, insulin resistance, TNF-α, IL-17, JAK-STAT, T cell receptors and other signaling pathways. Experimental verification showed that the key target genes were differentially expressed.CONCLUSION  This study reveals the active components of Platycodon grandiflorum in the treatment of NAFLD and its mechanism of action, it provides reference value for further research.