基于UPLC-Q-TOF-MS技术和网络药理学探讨配伍甘草降低附子心脏毒性的作用机制

Exploring the effect of licorice aconite on reducing cardiotoxicity based on UPLC-Q-TOF-MS and network pharmacology

关键词:  甘草  附子  心脏毒性  网络药理学

Key words:  licorice  aconite  cardiotoxicity  network pharmacology

摘要:

[目的] 采用超高效液相色谱-四极杆-飞行时间质谱（UPLC-Q-TOF-MS）技术分别测定甘草、附子单煎样品和合煎的化学成分，并基于网络药理学研究阐明以甘草配伍降低附子心脏毒性的作用机制。[方法] 采用液质联用技术对甘草、附子单煎液及合煎液的化学成分进行分析，探究配伍前后化学成分的变化。基于成分分析结果，通过TCMSP、David、DisGeNet等数据库对成分靶点以及心脏毒性靶点进行检索。制作Venn图，将共有靶点输入String数据库进行蛋白质相互作用（PPI）网络构建，并通过Cytoscape软件将网络可视化，最后进行基因本体论（GO）富集分析及Kyoto京都基因与基因组百科全书（KEGG）富集分析。[结果] 与自建数据库比较，共鉴定得到甘草单煎液化合物21种、附子单煎液化合物65种、甘草与附子合煎液化合物71种。基于液质联用技术，运用网络药理学构建中药-活性成分-作用靶点-疾病网络。分析结果表明甘草-附子和心脏毒性疾病的交集靶点共436个，包括蛋白激酶B1（AKT1）、GAPDH等。KEGG结果显示配伍甘草可能通过丝裂原活化蛋白激酶（MAPK）信号通路等降低附子心脏毒性。[结论]甘草配伍附子可以改变附子毒性成分，并作用于AKT1、GAPDH等靶点，通过MAPK、磷脂酰肌醇-3-激酶（PI3K）-蛋白激酶B（AKT）等信号通路达到降低毒性的效果。

Abstract:

[Objective] To use ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS） technology to determine the chemical components of licorice and aconite single decoction samples and combined decoction samples，and to elucidate the mechanism of licorice compatibility in reducing aconite cardiotoxicity based on network pharmacology. [Methods] The chemical compositions of single decoction and combined decoction of licorice radix and aconite radix were analysed by liquid chromatography-mass spectrometry technology to investigate the changes in chemical composition before and after the combination. Based on the results of the compositional analyses，component targets as well as cardiotoxicity targets were searched through databases such as TCMSP，David and DisGeNet. The Wayne diagram was produced，and the shared targets were entered into the String database for PPI network construction，and the network was visualised by Cytoscape software，and finally GO enrichment analysis and KEGG enrichment analysis were performed. [Results] Compared with the self-constructed database，21 compounds were identified from the single decoction of licorice root， 65 compounds from the single decoction of aconite root，and 71 compounds from the combined decoction of licorice root and aconite root. Based on liquid chromatography-mass technology，network pharmacology was applied to construct traditional Chinese medicine-active ingredient-target-disease networks，respectively. The analysis showed that there were 436 intersecting targets of licorice radix and aconite radix and cardiotoxicity disease，including AKT1，GAPDH and so on. KEGG results showed that the compatibility and detoxification of licorice radix may reduce the cardiotoxicity of aconite radix through Mitogen-Activated Protein Kinase（MAPK） signaling pathway. [Conclusion] The combination of licorice and aconite can change the toxic components of aconite，and act on AKT1，GAPDH and other targets to reduce the toxicity through MAPK，Phosphatidylinositol 3-Kinase-Protein Kinase B （PI3K-AKT） and other signaling pathways.