基于HPLC-Q-TOF MS/MS和网络药理学的参蓉蛹草片质量标志物研究

目的 应用高效液相色谱-四极杆飞行时间质谱法（HPLC-Q-TOF-MS/MS）建立参蓉蛹草片化学成分的检测方法，并结合网络药理学和分子对接方法分析其抗疲劳的网络调控机制、预测参蓉蛹草片的质量标志物。方法 运用HPLC-Q-TOF-MS/MS对参蓉蛹草片物质基础进行分析鉴定。通过PubChem获取所选化合物的结构，利用TCMSP数据库及SwissTargetPrediction获取作用靶点；通过GeneCards、TTD、OMIM数据库筛选与疲劳相关的疾病靶点，获得药物和疲劳的交集靶点；运用UniProt数据库校准靶点名称；利用STRING数据库构建交集基因蛋白质-蛋白质相互作用（PPI）网络，并运用Cytoscape 3.8.0软件对PPI网络进行拓扑分析；最后借助Metascape在线分析核心交集基因，进行基因本体（GO）功能富集分析、京都基因与基因组百科全书（KEGG）通路富集分析，并对分析结果进行可视化。结果 通过HPLC-Q-TOF MS/MS共得到参蓉蛹草片124个化学成分，利用网络药理学对筛选出的23个化学成分进行生物信息学分析，发现其可作用于195个靶点干预33条信号通路。分子对接结果显示，活性化合物可进入受体靶点活性空腔，与其形成的对接构象合理。结论 参蓉蛹草片可通过多成分、多靶点、多途径发挥抗疲劳作用，基于HPLC-Q-TOF MS/MS，结合网络药理学和分子对接方法初步将人参皂苷类、苯乙醇苷类预测为参蓉蛹草片的质量标志物。

Research on Quality Markers of Shenrong Yongcao Tablets Based on HPLC-Q-TOF-MS/MS and Network Pharmacology

Objective To establish a detection method for the chemical components of Shenrong Yongcao Tablets by high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS/MS), and analyze its anti-fatigue mechanism in combination with network pharmacology and molecular docking,to predicte the quality markers (Q-markers) of Shenrong Yongcao Tablets.Methods The basic materials of Shenrong Yongcao Tablets were analyzed and identified by HPLC-Q-TOF-MS/MS. The structures of the selected compounds were obtained by Pubchem,and the targets of action were obtained using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and SwissTargetPrediction. GeneCards,Therapeutic Target Database (TTD),and Online Mendelian Inheritance in Man (OMIM) were employed to screen fatigue-related disease targets,and the intersection targets of Shenrong Yongcao Tablets and diseases were obtained,which were calibrated through UniProt. STRING was used to construct the protein-protein interaction (PPI) network,and the topology of the PPI network was performed using Cytoscape 3.8.0. Finally,with the help of Metascape,Gene Ontology (GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis were carried out,and the results were visualized.Results A total of 124 chemical components of Shenrong Yongcao Tablets were obtained by HPLC-Q-TOF MS/MS,and 23 chemical components screened by network pharmacology acted on 195 targets and intervened in 33 signaling pathways. Molecular docking showed that the active compounds entered the active cavity of the receptor target,forming reasonable docking conformation.Conclusion Shenrong Yongcao Tablets can exert anti-fatigue effects through multi-component,multi-target and multi-pathway mechanism,and ginsenosides and phenylethanol glycosides are preliminarily predicted as Q-markers of Shenrong Yongcao Tablets by HPLC-Q-TOF MS/MS,network pharmacology and molecular docking.