中国药物警戒 ›› 2026, Vol. 23 ›› Issue (4): 426-431.
DOI: 10.19803/j.1672-8629.20260011

• 基础与临床研究 • 上一篇    下一篇

1-甲基-3-硝基-1-亚硝基胍体外诱导慢性萎缩性胃炎细胞模型建立研究

麦明欣1, 吕晓彤1, 孙丽新1, 王欣之1,*, 曹宇2#   

  1. 1中国药科大学多靶标天然药物全国重点实验室新药筛选与药效评价中心,江苏 南京 210009;
    2江苏大学附属人民医院消化内科,江苏 镇江 212002
  • 收稿日期:2026-01-05 出版日期:2026-04-15 发布日期:2026-04-15
  • 通讯作者: *王欣之,女,博士,副研究员,分子药理学与毒理学。E-mail: wangxz@cpu.edu.cn。#为共同通信作者。
  • 作者简介:麦明欣,女,在读硕士,分子药理学与毒理学。
  • 基金资助:
    国家自然科学基金资助项目（82574686、82073948）

An in vitro Model of MNNG-Induced Chronic Atrophic Gastritis

MAI Mingxin1, LYU Xiaotong1, SUN Lixin1, WANG Xinzhi1,*, CAO Yu2#   

  1. 1New Drug Screening and Pharmacodynamics Evaluation Center, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing Jiangsu 210009, China;
    2Department of Gastroenterology, the Affiliated People’ s Hospital of Jiangsu University, Zhenjiang Jiangsu 212002, China;
  • Received:2026-01-05 Online:2026-04-15 Published:2026-04-15

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摘要： 目的 探讨1-甲基-3-硝基-1-亚硝基胍（MNNG）体外诱导慢性萎缩性胃炎（CAG）不同病理阶段模型条件,为MNNG体外诱导CAG模型研究提供参考。方法 采用细胞增殖与毒性检测（CCK8）筛选MNNG作用于人胃黏膜上皮细胞系（GES-1细胞）浓度和时间;通过JC-1染色检测线粒体膜电位变化评估线粒体损伤程度;使用试剂盒检测丙二醛（MDA）、超氧化物歧化酶（SOD）以及乳酸脱氢酶（LDH）分别反映氧化应激水平与细胞凋亡程度;通过qRT-PCR检测胃型黏蛋白（SOX2、MUC5AC）、肠化生标志物（MUC2、KLF4、CDX2）及促炎细胞因子（TNF-α、IL-6、IL-8）mRNA表达;Western Blot检测CDX2蛋白表达;ELISA检测TNF-α分泌水平;并以阳性药维酶素（WMS）进行干预,评价模型构建效果。结果 20 μmol·L-1 MNNG处理GES-1细胞8 h可诱导早期CAG特征,表现为LDH释放增加、CDX2表达上调和TNF-α水平升高;处理24 h后,细胞呈现CAG伴不完全型肠化生表型,包括线粒体损伤、氧化应激加剧,细胞凋亡增加,胃型黏蛋白表达下调、肠化生标志物及促炎因子表达上调。WMS可显著逆转上述异常变化,提示模型建立成功。结论 20 μmol·L-1 MNNG处理GES-1细胞8 h可建立CAG模型,24 h可进一步诱导为CAG伴不完全肠化生模型,该模型较好模拟临床病理特征,为CAG机制研究及MNNG毒理学评价提供参考。

关键词: 慢性萎缩性胃炎, 肠化生, 人胃黏膜上皮细胞系（GES-1细胞）, 1-甲基-3-硝基-1-亚硝基胍（MNNG）, 体外模型

Abstract: Objective To screen the conditions for establishment of models representing different pathological stages of 1-methyl-3-nitro-1-nitrosoguanidine (MNNG)-induced chronic atrophic gastritis (CAG). Methods The cell counting kit-8 (CCK-8) assay was used to screen the concentration and time at which MNNG acted on the human gastric mucosal epithelial cell line (GES-1 cells). Mitochondrial membrane potential changes were detected via JC-1 staining to assess the severity of mitochondrial damage. Commercial assay kits were employed to measure levels of malondialdehyde (MDA), superoxide dismutase (SOD) and lactate dehydrogenase (LDH), which reflected oxidative stress and cell apoptosis, respectively. qRT-PCR was performed to detect the mRNA expression levels of gastric mucins (SOX2, MUC5AC), intestinal metaplasia markers (MUC2, KLF4, CDX2) and proinflammatory cytokines (TNF-α, IL-6, IL-8). Western blot was used to determine the protein expression of CDX2 while ELISA was applied to detect the secretory level of TNF-α. In addition, vitacoenzyme(WMS), a positive control drug, was used for interventions to evaluate the efficiency of the cell model. Results Eight hours of treatment of GES-1 cells with 20 μmol·L-1 MNNG induced signs of early CAG, manifested as increased LDH release, upregulated CDX2 expressions and elevated TNF-α levels. After 24 h of treatment, the cells exhibited the incomplete type of intestinal metaplasia, including mitochondrial damage, aggravated oxidative stress, increased cell apoptosis, downregulated expressions of gastric mucins, and upregulated expressions of intestinal metaplasia markers and proinflammatory cytokines. However, WMS significantly reversed the above abnormalities, indicating the applicability of the cell model. Conclusion Eight hours of treatment of GES-1 cells with 20 μmol·L-1 MNNG can help establish a CAG cell model, and 24 hours of treatment can lead to a model of CAG with the incomplete type of intestinal metaplasia. This cell model can well simulate the clinical pathology of the disease, thereby contributing to the mechanism research of CAG and the toxicological evaluation of MNNG.

Key words: Chronic Atrophic Gastritis, GES-1 Cells, Intestinal Metaplasia, 1-methyl-3-N'-nitro-1-nitrosoguanidine（MNNG）, in vitro Model

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