排毒清脂片的UPLC-Q-TOF-MS成分分析及网络药理学探索

Investigation of Paidu Qingzhi Tablets Based on UPLC-Q-TOF-MS and Network Pharmacology

目的:利用UPLC-Q-TOF-MS鉴定排毒清脂片的化学成分,并在此基础上开展网络药理学研究。方法:采用Waters ACQUITY UPLC HSS T3色谱柱(2.1 mm×100 mm,1.8μm),流动相0.1%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~17 min,95%~5%A;17~17.01 min,5%~95%A;17.01~20 min,95%A),流速0.3 m L·min-1,柱温40℃,使用电喷雾离子源(ESI),负离子模式采集数据,质谱扫描范围m/z 50~1200,通过对照品、相对分子质量、质谱裂解规律和文献信息鉴定排毒清脂片的化学成分;运用多个数据库检索化学成分对应靶点、排毒清脂片所治疾病靶点以及代谢通路,利用Cytoscape 3.7.1软件构建"药材-成分-靶点-通路-疾病"的可视化网络拓扑关系图。结果:从排毒清脂片甲醇提取物中鉴定出了33个成分,其中27种来源大黄,4种来源西洋参,2种来源麦冬。网络药理学研究表明其中31个化学成分的18个直接靶点,58个间接靶点及类固醇激素生物合成通路、花生四烯酸代谢通路、胰岛素信号通路等7条通路与排毒清脂片治疗高脂血症、痤疮、单纯性肥胖有较强的相关性。结论:UPLC-Q-TOF-MS技术可快速、高效对排毒清脂片进行化学成分定性分析,揭示了排毒清脂片的药效物质基础;网络药理学研究可辨识大黄、西洋参、麦冬3味中药的作用靶点及代谢通路,初步阐释排毒清脂片的分子作用机制,可为该制剂的质量控制与临床应用提供参考。

Objective:To identify the active constituents of Paidu Qingzhi tablets by UPLC-Q-TOF-MS technique,and to reveal its potential targets and molecular mechanisms by network pharmacology.Method:Chromatography separation was achieved on an ACQUITY UPLC HSS T3 column(2.1 mm×100 mm,1.8μm)with mobile phase consisted of 0.1%formic acid aqueous solution and acetonitrile for gradient elution(0-17 min,95%-5%A;17-17.01 min,5%-95%A;17.01-20 min,95%A),the flow rate was 0.3 m L·min-1,the column temperature was 40℃.Data acquisition was carried out in electrospray ionization(ESI)under the negative ion mode,the scanning range was 50-1200.Ingredients in Paidu Qingzhi tablets were identified according to reference substance,relative molecular weight,mass spectrometric cleavage rule and literature information.Multiple databases were used to retrieve the targets of these identified ingredients and related diseases treated by the tablets,and metabolic pathways.A visual network of"herbs-compounds-targets-pathways-diseases"was constructed by Cytoscape 3.7.1.Result:A total of 33 active compounds in methanol extract of Paidu Qingzhi tablets were identified by UPLC-Q-TOF-MS,including 27 compounds from Rhei Radix et Rhizoma,4 compounds from Panacis Quinquefolii Radix,2 compounds from Ophiopogonis Radix.Totally 18 direct targets and 58 indirect targets corresponding to 31 ingredients were obtained,7 metabolic pathways including steroid hormone biosynthesis pathway,arachidonic acid metabolic pathway,insulin signaling pathway were strongly correlated with the treatment of hyperlipidemia,acne and simple obesity by Paidu Qingzhi tablets.Conclusion:Ingredients of Paidu Qingzhi tablets are revealed by UPLC-Q-TOF-MS,based on these identified ingredients,targets and related metabolic pathways are visualized by network pharmacology.The current research can provide theoretical basis for quality control and clinical application of Paidu Qingzhi tablets.