An in vitro Model of MNNG-Induced Chronic Atrophic Gastritis

doi:10.19803/j.1672-8629.20260011

Abstract

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

CLC Number:

R994.11

MAI Mingxin, LYU Xiaotong, SUN Lixin, WANG Xinzhi, CAO Yu. An in vitro Model of MNNG-Induced Chronic Atrophic Gastritis[J]. Chinese Journal of Pharmacovigilance, 2026, 23(4): 426-431.

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URL: https://zgywjj.magtechjournal.com/EN/10.19803/j.1672-8629.20260011

https://zgywjj.magtechjournal.com/EN/Y2026/V23/I4/426

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