基于UPLC-Q-TOF/MS、网络药理学和分子对接模拟探讨黄连解毒汤治疗痛风性关节炎的药效物质及作用机制

Pharmacodynamic substances and mechanism of action of Huanglian Jiedu Decoction in the treatment of gouty arthritis:a study based on UPLC-Q-TOF/MS，network pharmacology，and molecular docking simulation

摘要

目的：利用超高效液相色谱-四极杆-飞行时间质谱（ultra performance liquid chromatography-quadrupole-time of flightmass spectrometry,UPLC-Q-TOF-MS）技术对黄连解毒汤（Huanglian Jiedu Decoction,HLJDD）的主要成分进行鉴定。并结合网络药理学和分子对接验证对HLJDD治疗痛风性关节炎（gouty arthritis,GA）中的潜在作用机制进行探讨。方法：通过UPLC-QTOF-MS技术在采集正、负离子模式下的数据，并结合对照品、相关文献及数据库检索，分析HLJDD的化学成分。利用网络药理学分析HLJDD治疗GA的潜在机制，筛选活性成分和GA的交集靶点，进一步富集分析和建立可视化网络，分子对接验证活性成分和交集靶点的结合能力。结果：通过UPLC-Q-TOF/MS方法鉴定出47个成分，采用网络药理学方法筛选得到HLJDD治疗GA的关键成分54个，交集靶点37个，PPI分析得到其中度值前10的关键靶点。GO功能富集分析得到20种生物过程、7种细胞组分、8种分子功能。KEGG通路富集分析得到96条干预GA相关通路，发现其治疗GA相关的通路主要涉及IL-17、TNF等炎症信号通路。分子对接进一步验证了HLJDD的关键成分与核心靶点结合能力良好。结论：已鉴定的HLJDD中的关键成分如黄柏碱、黄连碱、汉黄芩素、β-谷甾醇等可能通过调节IL-17、TNF通路中的多个核心靶点如PTSG2等来缓解GA，这为下一步深入研究作用机制提供了理论依据。

Abstract

Objective To identify the main components of Huanglian Jiedu Decoction（HLJDD） using ultra-high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry（UPLC-Q-TOF-MS），and to explore the potential mechanism of action of HLJDD in the treatment of gouty arthritis（GA） using network pharmacology and molecular docking methods. Methods We identified the chemical components of HLJDD by combining UPLC-Q-TOF-MS data acquired in both positive and negative ion modes with reference standards，relevant literature，and database searches. We analyzed the potential therapeutic mechanism of HLJDD for GA by using network pharmacology to determine the intersection targets between the active ingredients of HLJDD and GA for further enrichment analysis and visual network mapping. The binding affinity of the active ingredients with the intersection targets was validated through molecular docking. Results A total of 47 components were identified by UPLC-Q-TOF-MS； 54 key components of HLJDD for GA treatment and 37 intersection targets were determined by network pharmacology； and the top 10 key targets by Degree value were obtained by protein-protein interaction analysis. The Gene Ontology functional enrichment analysis revealed 20 biological processes，7 cellular components，and 8 molecular functions. The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis demonstrated 96 GA-related intervention pathways，in which inflammatory signaling pathways such as interleukin-17（IL-17） and tumor necrosis factor（TNF） were involved. Molecular docking verified that the key components of HLJDD had high binding affinity with the core targets. Conclusion The identified key components in HLJDD，such as phellodendrine，coptisine，wogonin，and β-sitosterol，may alleviate GA by regulating multiple core targets in the IL-17 and TNF pathways，such as PTSG2，which provides a theoretical basis for future investigation into the mechanism of action of HLJDD.

Graphical abstract

关键词

黄连解毒汤 / 化学成分 / 超高效液相色谱-四极杆-飞行时间串联质谱 / 网络药理学 / 分子对接

Key words

Huanglian Jiedu Decoction / chemical component / ultra-high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry / network pharmacology / molecular docking

基金资助

陕西省科技厅面上资助项目(2021JM-470)

陕西省教育厅重点资助项目(24JR051)

秦创原中医药产业创新聚集区资助项目(L2024-QCY-ZYYJJQ)