基于HPLC-Q-TOF-MS/MS与网络药理学小儿佛芍和中颗粒成分解析及治疗功能性腹痛药效物质基础及机制预测

Component analysis and pharmacological mechanism prediction of Xiaoer Foshao Hezhong Granules in treatment of functional abdominal pain based on HPLC-Q-TOF-MS/MS and network pharmacology

目的通过HPLC-Q-TOF-MS/MS分析小儿佛芍和中颗粒(XFHG)中的化学成分,依据解析的化学成分结合网络药理学预测其治疗功能性腹痛的药效物质基础,筛选出潜在功效成分,预测作用机制。方法采用HPLC-Q-TOF-MS/MS对XFHG进行化学成分分析,分别在正、负离子模式下扫描,结合对照品与中药系统药理学数据库与分析平台(TCMSP)、文献中的碎片离子信息、保留时间进行匹配,确认化学成分。筛选质谱解析得到的化学成分中口服生物利用度≥20%、类药性≥0.5的成分,同时纳入《中国药典》2020年版中规定的指标成分以及SwissADME数据库筛选得到的成分作为活性成分;通过TCMSP、Massbank、本草组鉴数据库、Genecards数据库及文献筛选,分别建立XFHG活性成分靶点数据库、功能性腹痛相关疾病的靶点数据库,筛选出共同靶点,导入网络可视化软件Cytoscape 3.9.1中,构建“XFHG-成分-功能性腹痛”靶点相互作用网络图,筛选出核心成分;共有靶点导入STRING11.5数据库中,构建蛋白质-蛋白质相互作用(PPI)网络图,并将得到的数据导入Cytoscape 3.9.1中,筛选出核心作用的靶点;将核心靶点导入DAVID数据库进行京都基因与基因组百科全书(KEGG)通路和基因本体(GO)富集分析。结果XFHG定性分析中共解析出96个化学成分,包括26个黄酮类、17个有机酸类、13个萜类、11个生物碱类、6个柠檬苦素类、8个苯丙素类、4个苯乙醇苷类及11个其他类;筛选出发挥治疗功能性腹痛作用的核心成分15个,来源于6味药材,包括小檗碱、芍药苷、连翘酯苷A、连翘苷、吴茱萸碱、吴茱萸次碱等;XFHG治疗功能性腹痛的核心靶点有TNF、AKT1、IL6、IL-1β、VEGFA、TP53、CASP3、PTGS2、CAT、MAPK3;KEGG通路分析共得到109条通路,免疫应答、炎症反应相关的信号通路排名靠前;GO富集分析得到生物过程(BP)条目145个、细胞组分(CC)条目10个、分子功能(MF)条目9个,富集值排名靠前的条目涉及脂多糖介导的信号通路、糖皮质激素反应、一氧化氮生物合成过程的正向调控等与炎症反应相关的生物过程。结论根据HPLC-Q-TOF-MS/MS解析的化学成分结合网络药理学预测了XFHG治疗功能性腹痛的潜在功效成分,预测其作用机制与免疫调节、抗炎相关。

Objective To Analyze the chemical composition of Xiaoer Foshao Hezhong Granules(XFHG)by HPLC-Q-TOF-MS/MS,then predicting the pharmacodynamic substance basis of treating functional abdominal pain based on its chemical composition and network pharmacology,checking out the potential efficacy ingredients and mechanism of action.Methods HPLC-Q-TOF-MS/MS was used to analyze the chemical composition of XFHG,which were scanned in positive and negative modes respectively.The chemical composition was confirmed by matching the reference substance with the fragment ion information and retention time in the Systems Pharmacology Database and Analysis Platform of traditional Chinese medicine(TCMSP)and literatures.The chemical components obtained through mass spectrometry analysis with oral bioavailability≥20%and drug like properties≥0.5 were selected,and the target components specified in the 2020 edition of the Chinese Pharmacopoeia and the components screened from the SwissADME database were included as active ingredients.By using TCMSP,Massbank,Herbal Pharmacopoeia Database,Genecards Database,and literature screening,we established target databases for active ingredients of XFHG,as well as target databases for functional abdominal pain related diseases.We screened common targets and imported them into the network visualization software Cytoscape 3.9.1 to construct a target interaction network diagram for"XFHG-components-functional abdominal pain",and screened the core components.Import common targets into the STRING 11.5 database,construct a protein protein interaction(PPI)network diagram,and import the obtained data into Cytoscape 3.9.1 to screen out the core targets.The core targets were imported into the DAVID database for Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway and Gene Ontology(GO)enrichment analysis.Results There were 96 chemical components analyzed by qualitative analysis of XFHG,including 26 flavonoids,17 organic acids,13 terpenoids,11 alkaloids,6 lemon,8 phenylpropanoids,4 benzylethanol side and 11 other classes.Screening out 15 core components that play a therapeutic role in functional abdominal pain,sourced from six medicinal herbs,including berberine,paeoniflorin,forsythrin A,forsythrin,evodiamine,rutaecarpine,etc.The core targets of XFHG for treating functional abdominal pain include TNF,AKT1,IL6,IL-1β,VEGFA,TP53,CASP3,PTGS2,CAT,and MAPK3.A total of 109 pathways were identified through KEGG pathway analysis,with immune response and inflammatory response related signaling pathways ranking high.GO enrichment analysis yielded 145 entries for biological processes(BP),10 entries for cellular components(CC),and nine entries for molecular functions(MF).The entries with the highest enrichment values involved biological processes related to inflammation,such as lipopolysaccharide-mediated signaling pathways,glucocorticoid responses,and positive regulation of nitric oxide biosynthesis processes.Conclusion Based on the chemical components analyzed by HPLC-Q-TOF-MS/MS combined with network pharmacology,the potential efficacy components of XFHG in the treatment of functional abdominal pain were predicted,and its mechanism of action was predicted to be related to immune regulation and anti-inflammatory effects.