中国药物警戒 ›› 2026, Vol. 23 ›› Issue (10): 1115-1122.
DOI: 10.19803/j.1672-8629.20260697

• 蒙药药效物质基础与作用机制研究专栏（二） • 上一篇    下一篇

小秦艽花异荭草苷经MAPK通路调控COPD相关气道炎症的体外机制研究

宋汶轩1,2, 周树宏2, 张文雪1,2, 王宁2, 郭琪2,3, 马奥2,3, 王敏杰2,4#, 伊乐泰2,5*   

  1. 1内蒙古医科大学药学院,内蒙古 呼和浩特 010110;
    2内蒙古医科大学中药（蒙药）质量研究与药效评价重点实验室,内蒙古 呼和浩特 010059;
    3包头医学院药学院,内蒙古 包头 014040;
    4内蒙古医科大学基础医学院,内蒙古 呼和浩特 010110;
    5内蒙古医科大学民族医药创新中心,内蒙古 呼和浩特 010110
  • 收稿日期:2026-08-24 出版日期:2026-10-15 发布日期:2026-10-16
  • 通讯作者: *伊乐泰,男,硕士,教授,中蒙药活性物质基础研究。E-mail: yiletai@126.com。#为共同通信作者。
  • 作者简介:宋汶轩,女,在读硕士,中蒙药活性物质基础研究。
  • 基金资助:
    国家自然科学基金资助项目（82360828）

Isoorientin from Gentiana dahurica Fisch. Flower regulates COPD-related airway inflammation via MAPK pathway: an in vitro study of mechanisms

Song Wenxuan1,2, Zhou Shuhong2, Zhang Wenxue1,2, Wang Ning2, Guo Qi2,3, Ma Ao2,3, Wang Minjie2,4#, Yi Letai2,5*   

  1. 1School of Pharmacy, Inner Mongolia Medical University, Hohhot Inner Mongolia 010110, China;
    2Key Laboratory of Quality Research and Efficacy Evaluation of Traditional Chinese Medicine (Mongolian medicine), Inner Mongolia Medical University, Hohhot Inner Mongolia 010059, China;
    3School of Pharmacy, Baotou Medical College, Baotou Inner Mongolia 014040, China;
    4School of Basic Medicine, Inner Mongolia Medical University, Hohhot Inner Mongolia 010110, China;
    5Medical Innovation Center for Nationalities, Inner Mongolia Medical University, Hohhot Inner Mongolia 010110, China
  • Received:2026-08-24 Online:2026-10-15 Published:2026-10-16

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摘要： 目的 探讨小秦艽花活性成分异荭草苷（Isoorientin,ISO）对慢性阻塞性肺疾病（Chronic Obstructive PulmonaryDisease,COPD）相关气道炎症的改善作用及其分子机制。方法 采用高效液相色谱串联三重四级杆质谱（LC-MS/MS）法测定小秦艽花中异荭草苷、龙胆苦苷、马钱苷酸的含量。采用脂多糖（LPS）5 μg·mL-1诱导人支气管上皮BEAS-2B细胞构建COPD相关气道炎症体外细胞模型。采用CCK-8法检测不同浓度、不同干预时间ISO对BEAS-2B细胞活力的影响,筛选安全给药条件并评价其对炎症损伤细胞的保护作用。采用RT-qPCR法检测炎症相关因子mRNA表达水平。通过转录组测序筛选差异基因及核心富集通路。采用RT-qPCR和Western Blot法检测MAPK通路关键基因与蛋白表达水平。借助分子对接模拟分析,评估异荭草苷与通路核心蛋白的结合活性。结果 ISO是小秦艽花中含量较高的代表性活性成分。细胞实验筛选获得ISO安全干预条件为作用12 h浓度5 μmol·L-1。与对照组比较,模型组细胞活力显著降低（P<0.001）;与模型组比较,5 μmol·L-1 ISO可显著提高炎症损伤细胞活力（P<0.01）。模型组中IL-6、IL-1β、IL-8、TNF-α、MMP-9、CXCL5这些炎症相关基因显著高表达（P<0.001）,与模型组比较,ISO可显著下调上述炎症因子的mRNA表达（P<0.01、P<0.001）。转录组富集分析提示MAPK通路可能为ISO发挥抗炎作用的重要途径,模型组中差异基因EREG、MAPK8（JNK1）、FOS表达显著上调（P<0.01、P<0.001）,EREG、c-Fos蛋白表达升高（P<0.01）,p-JNK/JNK比值增加（P<0.01）;ISO干预可显著下调上述基因与蛋白的异常高表达（P<0.01、P<0.001）,降低p-JNK/JNK比值（P<0.01）,从而抑制LPS诱导的MAPK通路过度激活。分子对接结果显示,ISO与EREG、JNK1、c-Fos蛋白的结合能均低于-5 kcal·mol-1,提示ISO与上述核心蛋白存在较好的结合能力,从分子模拟层面表明ISO可能通过结合通路关键蛋白进而影响MAPK通路的活化状态。结论 小秦艽花中的活性成分ISO可能通过与MAPK通路关键蛋白相互作用,抑制该通路异常激活,进而减少气道上皮炎症因子的释放,从而减轻LPS诱导的COPD相关气道炎症损伤。

关键词: 小秦艽花, 异荭草苷, 慢性阻塞性肺疾病（COPD）, COPD相关气道炎症, 体外细胞实验, 人支气管上皮BEAS-2B细胞, 转录组学

Abstract: Objective To explore the role of ISO, an active component of Gentiana dahurica Fisch. Flower in attenuating airway inflammation related to chronic obstructive pulmonary disease (COPD) and the molecular mechanism. Methods ISO, gentiopicroside and loganic acid in Gentiana dahurica Fisch. Flower were quantified via LC-MS/MS. Lipopolysa-ccharide (LPS, 5 μg·mL-1) was used to establish an in-vitro COPD-related inflammatory model in BEAS-2B human bronchial epithelial cells. CCK-8 assay was performed to screen the safe ISO dosage and evaluate its effect of cytoprotection. RT-qPCR was adopted to detect mRNA levels of inflammatory mediators. Transcriptome sequencing was employed to screen differentially expressed genes and key enriched pathways. RT-qPCR and Western blot were used to measure the expression levels of core MAPK-related genes and proteins. Molecular docking was conducted to predict the binding affinity between isoorientin and target proteins. Results ISO was the leading active component of Gentiana dahurica Fisch. Flower. The safe intervention condition of ISO was 5 μmol•L⁻¹ and 12 hours of intervention. Compared with the control group, LPS markedly reduced cell viability (P<0.001) while 5 μmol·L-1 ISO restored cell viability in the model group (P<0.01). The mRNA levels of IL-6, IL-1β, IL-8, TNF-α, MMP-9 and CXCL5 were significantly elevated in LPS-treated cells (P<0.001), and ISO markedly suppressed these up-regulated transcripts (P<0.01, P<0.001). Transcriptome enrichment identified the MAPK pathway as a major anti-inflammatory target. LPS up-regulated EREG, MAPK8 (JNK1) and FOS at the mRNA level (P<0.01, P<0.001), increased EREG, c-Fos protein abundance and elevated the p-JNK/JNK ratio (P<0.01), but ISO reversed these alterations and restrained excessive MAPK activation (P<0.01, P<0.001). Molecular docking suggested that binding energies between ISO and EREG, JNK1 and c-Fos were all less than -5 kcal·mol-1, suggesting favorable binding potentials. Conclusion ISO derived from Gentiana dahurica Fisch. Flower may inhibit aberrant MAPK signaling, reduce pro-inflammatory mediator release and alleviate LPS-provoked airway inflammatory injury related to COPD by interacting with key MAPK-related proteins.

Key words: Gentiana dahurica Fisch. Flower, Isoorientin, Chronic Obstructive Pulmonary Disease (COPD), COPD Related Airway Inflammation, In vitro Cell Experiment, BEAS-2B Human Bronchial Epithelial Cells, Transcriptomics

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