南方医科大学学报

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (8): 1654-1662.doi: 10.12122/j.issn.1673-4254.2025.08.10

• • 上一篇    

2，6-二甲氧基-1，4-苯醌通过抑制NLRP3炎症小体活化改善葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎

刘辰菲1,2(), 张玮1,2, 曾尧1,2, 梁艳2, 王梦婷2, 张明芳2, 李新元2, 王凤超1, 杨燕青1,2()   

  1. 1.蚌埠医科大学第一附属医院检验科，安徽 蚌埠 233004
    2.蚌埠医科大学慢性疾病免疫学基础与临床安徽省重点实验室，安徽 蚌埠 233030
  • 收稿日期:2025-03-25 出版日期:2025-08-20 发布日期:2025-09-05
  • 通讯作者: 杨燕青 E-mail:2628819350@qq.com;yyqing@mail.ustc.edu.cn
  • 作者简介:刘辰菲，在读硕士研究生，E-mail: 2628819350@qq.com
  • 基金资助:
    国家自然科学基金(82071775);安徽省科研编制计划优秀青年科研项目(2022AH030140);蚌埠医学院2024年度研究生科研创新计划项目(Byycx24101);蚌埠医学院2024年度研究生科研创新计划项目(Byycx24030);2023年国家级大学生创新创业训练计划项目(202310367020);蚌医一附院高水平科技创新团队资助(BYYFY2022TD001)

2,6-dimethoxy-1,4-benzoquinone alleviates dextran sulfate sodium-induced ulcerative colitis in mice by suppressing NLRP3 inflammasome activation

Chenfei LIU1,2(), Wei ZHANG1,2, Yao ZENG1,2, Yan LIANG2, Mengting WANG2, Mingfang ZHANG2, Xinyuan LI2, Fengchao WANG1, Yanqing YANG1,2()   

  1. 1.Clinical Laboratory, First Affiliated Hospital of Bengbu Medical University, Bengbu 233004, China
    2.Anhui Provincial Key Laboratory of Immunology in Chronic Diseases, Bengbu Medical University, Bengbu 233030, China
  • Received:2025-03-25 Online:2025-08-20 Published:2025-09-05
  • Contact: Yanqing YANG E-mail:2628819350@qq.com;yyqing@mail.ustc.edu.cn
  • Supported by:
    National Science Foundation of China(82071775)

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摘要：

目的 探究2，6-二甲氧基-1，4-苯醌（DMQ）对葡聚糖硫酸钠（DSS）诱导的小鼠溃疡性结肠炎的作用及机制。 方法 将18只雄性C57BL/6J小鼠随机分为3组：WT组、DSS组和DMQ干预组，6只/组。DSS组和DMQ干预组小鼠均采用DSS诱导建立溃疡性结肠炎模型，DMQ干预组小鼠给予20 mg/kg的DMQ腹腔注射，DSS组小鼠给予无菌PBS腹腔注射。监测各组小鼠的体质量变化、疾病活动指数（DAI）、结肠长度、脾脏质量、HE染色、组织学评分，探究DMQ对DSS诱导溃疡性结肠炎小鼠是否具有缓解作用。流式细胞术检测小鼠肠系膜淋巴结和脾脏组织中Th17细胞和分泌Th1型细胞因子IFN-γ的CD8+ T细胞比例变化，Western blotting检测紧密连接蛋白（Occludin、ZO-1）和NLRP3炎症小体活化相关蛋白（成熟的Caspase-1，即p20）以及ELISA检测炎症因子（IL-1β、TNF-α）。小鼠骨髓来源巨噬细胞（BMDM）由脂多糖（LPS）预处理、DMQ干预后，应用尼日利亚菌素（Nigericin）活化经典途径NLRP3炎症小体；以及采用人外周血单个核细胞（PBMC），应用尼日利亚菌素（Nigericin）活化经典途径NLRP3炎症小体，LPS活化替代途径NLRP3炎症小体，通过Western blotting、ELISA和流式细胞术等技术，检测了炎症相关蛋白表达、炎性因子分泌及细胞焦亡情况。 结果 与DSS组相比，DMQ治疗可缓解DSS小鼠的体质量下降（P<0.001）、结肠长度缩短（P<0.05）、脾脏质量增加（P<0.001）和结肠炎症（P<0.05）。在DMQ治疗组小鼠肠系膜淋巴结和脾脏组织中，流式细胞术结果显示：T细胞中Th17细胞比例和分泌Th1型细胞因子IFN-γ的CD8+T细胞比例降低（P<0.05），Western blotting结果显示：Occludin和ZO-1的蛋白表达升高，而Caspase-1的蛋白表达降低；进一步实验结果显示：DMQ可有效抑制小鼠骨髓来源巨噬细胞（BMDM）中经典途径NLRP3炎症小体活化和人外周血单个核细胞（PBMC）中经典途径NLRP3炎症小体和替代途径NLRP3炎症小体活化（P<0.05）以及Caspase-1依赖性细胞焦亡。 结论 DMQ通过抑制NLRP3炎症小体活化发挥对DSS诱导的小鼠溃疡性结肠炎的保护作用。

关键词: 2，6-二甲氧基-1，4-苯醌, NLRP3炎症小体, 溃疡性结肠炎, 天然产物

Abstract:

Objective To investigate the therapeutic mechanism of 2,6-dimethoxy-1,4-benzoquinone (DMQ) for alleviating dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice. Methods Eighteen male C57BL/6J mice were equally randomized into control group, DSS group and DMQ treatment group. In DSS and DMQ groups, the mice were treated with DSS in drinking water to induce UC, and received intraperitoneal injections of sterile PBS or DMQ (20 mg/kg) during modeling. The changes in body weight, disease activity index (DAI), colon length, spleen weight, and colon histological scores of the mice were examined, and the percentages of Th17 and IFN-γ+ CD8+ T cells in the mesenteric lymph nodes and spleen were analyzed using flow cytometry. The expressions of tight junction proteins (Occludin and ZO-1), proteins associated with inflammasome activation (caspase-1 and p20), IL-1β and TNF-α in the colon tissues were detected using Western blotting or ELISA. In the cell experiment, mouse bone marrow-derived macrophages (BMDMs) primed with lipopolysaccharide (LPS) were treated with DMQ, followed by stmulation with nigericin to activate the classical NLRP3 inflammasome pathway. In cultured human peripheral blood mononuclear cells (PBMCs) treated with either LPS alone or LPS plus nigericin, the effects of DMQ on inflammasome activation, pyroptosis, and cytokine release were evaluated via Western blotting, ELISA, and flow cytometry. Results In DSS-treated mice, DMQ treatment significantly alleviated DSS-induced body weight loss, colon shortening, spleen enlargement, and colon inflammation. The DMQ-treated mice showed significantly reduced percentages of Th17 cells and IFN-γ+ CD8+ T cells in the mesenteric lymph nodes and spleen, with increased occludin and ZO-1 expressions and decreased caspase-1 expression in the colon tissue. DMQ obviously inhibited classical NLRP3 inflammasome activation in mouse BMDMs and both the classical and alternative pathways of NLRP3 activation in human PBMCs, causing also suppression of caspase-1-dependent pyroptosis. Conclusion DMQ ameliorates DSS-induced UC in mice by inhibiting NLRP3 inflammasome activation.

Key words: 2,6-dimethoxy-1,4-benzoquinone, NLRP3 inflammasome, ulcerative colitis, natural products