南方医科大学学报

南方医科大学学报 ›› 2023, Vol. 43 ›› Issue (6): 879-888.doi: 10.12122/j.issn.1673-4254.2023.06.02

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东革阿里抗炎的物质基础及其作用机制：基于UPLC-Q-TOF-MS/MS和网络药理学方法

刘 芳，张远芳，刘 鹏，刘佳敏，刘思妤，王俊杰   

  1. 湘南学院药学院，湖南 郴州 423000
  • 出版日期:2023-06-20 发布日期:2023-07-06

UPLC-Q-TOF-MS/MS combined with network pharmacology for exploring anti-inflammatory mechanism of Eurycoma longifolia

LIU Fang, ZHANG Yuanfang, LIU Peng, LIU Jiamin, LIU Siyu, WANG Junjie   

  1. College of Pharmacy, Xiangnan University, Chenzhou 423000, China
  • Online:2023-06-20 Published:2023-07-06

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摘要： 目的 阐明东革阿里抗炎活性的物质基础及作用机制。方法 利用二甲苯致耳肿胀和脂多糖致急性肺炎模型筛选东革阿里的活性部位；采用UPLC-Q-TOF-MS/MS技术鉴定活性部位的化学组成，结合SwissADME、SwissTargetPrediction、Genecards等数据库筛选东革阿里抗炎的潜在作用靶点，String数据库绘制蛋白互作（PPI）网络图，Cytoscape软件做网络拓扑分析并筛选核心靶点，通过Metascape数据库对核心靶点进行富集分析，使用Autodock软件进行核心成分和靶点进行分子对接，Pymol软件将对接结果可视化；采用脂多糖诱导的RAW264.7细胞炎症模型验证东革阿里抗炎作用机制，通过Griess试剂法检测NO的水平，Western blot法检测MAPK1、JAK2及STAT3蛋白的表达水平。结果 东革阿里75%提取物（ELE）为活性部位，该部位共鉴定出37个化学成分，化合物和疾病共有靶点541个，涉及JAK/STAT3、cAMP等信号通路，筛选出12个指标性成分，指标性成分与涉及信号通路上的2个核心靶点进行分子对接，对接结果良好；细胞验证实验显示ELE的低、中、高剂量组均能显著降低细胞NO的释放量，中剂量可以显著降低JAK2和STAT3的表达。结论 东革阿里抗炎活性的物质基础为苦木素及生物碱类成分，其抗炎作用的机制可能与靶向JAK2、STAT3调控JAK/STAT3信号通路有关。

关键词: 东革阿里；抗炎；活性基础；网络药理学；分子对接

Abstract: Objective To explore the mechanisms that mediate the anti-inflammatory activity of Eurycoma longifolia. Methods Kunming mouse models of xylene-induced ear swelling and lipopolysaccharide (LPS)-induced acute pneumonia were used to compare the anti- inflammatory activities of aqueous and ethanol extracts of Eurycoma longifolia. UPLC-Q-TOF-MS/MS was used to identify the chemical composition in the ethanol extract of Eurycoma longifolia, based on which the potential anti-inflammatory targets of Eurycoma longifolia were screened using the databases including SwissADME, SwissTargetPrediction, and Genecards. The String database was used to generate the protein-protein interaction (PPI) network, and Cytoscape was used for network topology analysis and screening the core targets. The enrichment of the core targets was analyzed using Metascape database, the core components and targets were docked with Autodock software, and the docking results were visualized using Pymol software. In a RAW264.7 cell model of LPS-induced inflammation, the Griess reagent was used to measure NO level, and Western blotting was performed to detect the expression levels of MAPK1, JAK2, and STAT3 proteins to verify the anti- inflammatory mechanism of Eurycoma longifolia. Results The ethanol extract (75% ) of Eurycoma longifolia (ELE) was the active site, which contained a total of 37 chemical components. These chemical compounds and diseases had 541 targets, involving the JAK/STAT3, cAMP and other signaling pathways. Twelve indicator components were identified, which all showed good results of molecular docking with two core targets involved in the signaling pathways. In the cell validation experiment, treatment of the cells with low-, medium-, and high-dose ELE significantly reduced NO release in the cells, and ELE at the medium dose significantly decreased the cellular expressions of JAK2 and STAT3. Conclusion The anti-inflammatory activity of Eurycoma longifolia is attributed primarily to its active ingredients bitter lignin and alkaloids, which may regulate the JAK/STAT3 signaling pathway by targeting JAK2 and STAT3.

Key words: Eurycoma longifolia Jack; anti-inflammation; pharmacological activity; network pharmacology; molecular docking