糖尿病（DM）及其相关并发症是一种全球性流行病，其特征为高发病率和死亡率。然而，对于糖尿病肠病（DE）及其潜在机制知之甚少。本研究从实验鼠中提取肠道上皮干细胞（IESCs），通过RT-PCR、蛋白质印迹、免疫组化检测了优势的N6甲基腺苷（m6A）相关酶的水平。通过沉默实验证实了Mettl14在DM期间肠道上皮细胞（IECs）异常分化中的作用。通过RT-PCR、MeRIP和生物信息学分析，确认了Mettl14的下游靶标。通过生物信息学分析、RT-PCR和蛋白质印迹进一步分析了DM小鼠的IECs中与分化相关的基因。本研究发现，在db/db小鼠中Mettl14和m6A的水平高于对照组小鼠。而Mettl14的过表达与DM中IECs的异常分化相关。此外，Mettl14是IESCs鉴别和在DM状态下形成器官类家族的主要决定性因子。机制上，我们发现Mettl14的候选结合靶标是Fzd2 mRNA，且影响Fzd2的稳定性。此外，在DM状态下观察到Mettl14的下调能够通过调节Fzd2的m6A修饰而减轻IECs的异常分化。综上所述，我们的结果提供了Mettl14在DM状态下IESCs分化中的重要作用的确凿证据。© 2023 Wiley Periodicals LLC.

Diabetes mellitus (DM) and its related complications are a global epidemic characterized by high morbidity and mortality. However, little is known about diabetic enteropathy (DE) and its the potential underlying mechanism. Intestinal epithelial stem cells (IESCs) were harvested from experimental mice, and the levels of dominant N6-methyladenosine (m6 A)-related enzyme were detected by RT-PCR, Western blotting, immunohistochemistry. The role of Mettl14 in the abnormal differentiation of intestinal epithelial cells (IECs) during DM was confirmed by knockdown experiments. RT-PCR, MeRIP, and bioinformatics analysis were carried out to confirm the downstream target of Mettl14. Through bioinformatics analysis, RT-PCR, and Western blotting, we further analyzed the differentiation-related gene in the IECs from mice with DM. In this study, the levels of Mettl14 and m6 A were higher in db/db mice than that in control mice. And abnormal differentiation of IECs in DM was associated with Mettl14 overexpression. Additionally, Mettl14 is a major determinant of IESCs identity and organoid-forming upon DM state. Mechanistically, we revealed that the candidate binding target of Mettl14 was Fzd2 mRNA and affected Fzd2 stability. Moreover, Mettl14 downregulation was observed to attenuate the abnormal differentiation of IECs through modulating Fzd2 m6A modification in DM state. Together, our results provide definitive evidence for the essential role of Mettl14 in differentiation of IESCs in DM state.© 2023 Wiley Periodicals LLC.