基于UPLC-Q-TOF-MS结合网络药理学的黄褐毛忍冬保肝活性成分及其潜在靶点研究

Chemical composition identification of hepatoprotection and potential targets by L.fulvotomentosa Hsu et S.C.Cheng based on integrated UPLC-Q-TOF-MS and network pharmacology

目的:系统研究黄褐毛忍冬的主要化学成分,基于网络药理学探讨其保肝活性成分,为其药效物质研究提供一定的参考依据。方法:利用超高效液相色谱飞行时间质谱联用技术(UPLC-Q-TOF-MS)对黄褐毛忍冬的主要化学成分进行分析,根据化合物的一级、二级质谱信息,并与标准品或参考文献进行比对,对黄褐毛忍冬的化学信息进行快速识别,利用Molinspiration平台预测药物分子的吸收情况。通过Swiss Target Prediction和Similarity ensemble approach(SEA)数据库检索黄褐毛忍冬化学成分对应靶点,与GeneCards数据库中急性肝损伤靶点进行比对,筛选黄褐毛忍冬保肝作用的靶点,并利用KOBAS 3.0数据库对核心靶点进行通路富集分析,采用Cytoscape 3.7.1软件构建“化学成分—靶点—通路”的可视化网络拓扑关系图。结果:黄褐毛忍冬提取物主要含有57种成分,利用Molinspiration平台筛选出24种可能被吸收的化学成分,其作用于319个急性肝损伤靶点。其中隐绿原酸、木犀草苷、绿原酸、新绿原酸、山柰酚、槲皮苷、齐墩果酸、栀子苷、断马钱子苷半缩醛内酯等主要化学成分可能作用于碳酸酐酶2(CA2)、醛糖还原酶1(AKR1B1)、碳酸酐酶1(CA1)、碳酸酐酶9(CA9)、血管内皮生长因子A(VEGFA)、白介素2(IL-2)、内质网氨肽酶基因1(ERAP1)、成纤维细胞生长因子1(FGF1)、成纤维细胞生长因子2(FGF2)、醛氧化酶(ALOX)、腺苷受体A1(ADORA1)、碳酸酐酶4(CA4)等靶点,富集于PI3K-Akt信号通路、MAPK信号通路、TNF信号通路、Toll受体信号通路、PPAR信号通路、Wnt信号通路、Notch信号通路等。结论:采用UPLC-Q-TOF-MS结合网络药理学的方法,初步揭示了黄褐毛忍冬的化学组成及保肝的潜在作用机制。

Objective:To systematically identify the chemical components and explore the potential mechanisms of hepatoprotection by L.fulvotomentosa Hsu et S.C.Cheng(LFH),and provide evidences for figuring out pharmacodynamic substances.Methods:First,the main chemical components of LFH were analyzed by UPLC-Q-TOF-MS,and quickly identified by standards or references based on the MS/MS spectrometry information of all potential compounds.The absorption efficacy of drugs was predicted by Molinspiration software.Then,Swiss Target Prediction and SEA database were used to retrieve the target of the chemical components of LFH,and the acute liver injury related targets were also screened by GeneCards databases and overlapping proteins were selected as acute liver injury targets of LFH.Furthermore,the target was obtained from KOBAS 3.0 database,and the“component-target-pathway”network model was established by Cytoscape 3.7.1 software.Results:Fifty-seven chemical constituents of LFH were screened out,among which 24 absorbed chemical components were selected by Molinspiration software and acted on 319 acute liver injury targets.Among,the chemical components,such as Cryptochlorogenic acid,Cynaroside,Chlorogenic acid,Neochlorogenic acid,Kaempferol,Quercetin,Oleanolic acid,Gardenoside,and Vogelosidemay act on the protein targets of CA2,AKR1B1,CA1,CA9,VEGFA,IL-2,ERAP1,FGF1,FGF2,ALOX,ADORA1 and CA4.The KEGG pathway analysis showed that LFH was directly or indirectly engaged in the signal pathways of PI3K-Akt,MAPK,TNF,Toll-like receptor,PPAR,Wnt,Notch and so on.Conclusion:UPLC-Q-TOF-MS combined with network pharmacology may be used to preliminarily clarify the chemical composition and reveal potential mechanisms of LFH.