中国药理学与毒理学杂志2025，Vol.39Issue(6)：444-452,9.DOI:10.3867/j.issn.1000-3002.2025.08282
乙醇诱导胃黏膜损伤模型小鼠胃干细胞的分化作用研究
Differentiation effect of gastric stem cells in ethanol-induced gastric mucous injury in mice
张林丽 1田书婷 1刘芮希 2刘莉1
作者信息
- 1. 中国医药工业研究总院,上海 200437||上海市生物物质成药性评价专业技术服务平台,上海 200437||上海医药工业研究院有限公司,先导物成药性研究全国重点实验室,上海 200437
- 2. 西安医学院临床医学院,陕西 西安 710021
- 折叠
摘要
Abstract
OBJECTIVE To study the effects of ethanol on markers of gastric stem cells and epithe-lial cells,and explore the related signal pathways for stem cells differentiation in a mouse gastric mucous injury model.METHODS Male C57BL/6 mice were randomly divided into normal control and ethanol groups.The mice in the control group were given normal drinking water while those in the ethanol group were gavaged with 10 ml·kg-1 50%(V/V)ethanol on day 1,and drinking water containing 10%(V/V)ethanol was given on day 2-9.On the 10th day,stomach tissues were collected.HE staining was used to detect pathological changes in the stomach.Immunohistochemistry(IHC)staining was used to detect changes of such cell markers as mucin 5AC,H+/K+ATPase β and pepsin C.Immunofluorescence(IF)staining was employed to analyze changes in expression of cell markers such as H+/K+ATPase β,pepsin C and gastrin.ELISA assay was used to measure gastrin,somatostatin and interleukin 1β(IL-1β)concentrations in gastric tissue homogenates.Flow cytometry was adopted to measure the number of leucine rich repeat containing G protein-coupled receptor 5 positive(LGR5+)stem cells in gastric glands.Organoids was constructed to characterize stem cell activity.RNA sequencing and bioinformatics analysis were conducted to analyze the inflammatory pathways and differentiation signaling pathways during mice gastric mucous injury.RESULTS H&E results showed multifocal necrosis of the mucosal layer appeared in the ethanol group,accompanied by pyknosis,lysis and detachment of mucosal epithelial cells and gastric gland cells.IHC results showed decreased expressions of mucin 5AC and increased expressions of H+/K+ATPase β and pepsin C.IF results revealed increased expressions of H+/K+ATPase β,pepsin C,and gastrin after ethanol treatment.ELISA results demonstrated significant increases in gastrin,somatostatin and IL-1β levels in gastric tissues of the ethanol group.Flow assay results suggested that the number of LGR5+stem cells significantly decreased in ethanol treated gastric tissues.Stem cells from stomach tissues treated with ethanol did not grow into organoids.RNA sequencing and bioinformatics analyses revealed enrichment of TNF,NF-κB and Notch pathways in the ethanol group.CONCLUSION Administration of 50%ethanol solution on day 1,followed by continuous admin-istration of 10%ethanol solution on day 2-9 can induce histopathological injury to the gastric gland and stem cells,and an increase in epithelial cells.These changes may be related to the up-regulation of inflammatory pathways and Notch signaling pathways triggered by ethanol.关键词
干细胞/免疫荧光/类器官Key words
stem cell/immunofluorescence/organoid分类
医药卫生引用本文复制引用
张林丽,田书婷,刘芮希,刘莉..乙醇诱导胃黏膜损伤模型小鼠胃干细胞的分化作用研究[J].中国药理学与毒理学杂志,2025,39(6):444-452,9.
