我们旨在研究保守的哺乳动物NAD +依赖的蛋白去乙酰化酶SIRT1如何调节肠内分泌细胞(EEC)的数量。EEC有益于代谢，增加它们可能治疗2型糖尿病和肥胖。我们使用了特异性在肠上皮(VilKO，villin-Cre + 和Sirt1flox/flox小鼠)或肠内分泌细胞祖细胞 (EEPCs) (NgnKO，neurogenin3(ngns)-Cre + 和Sirt1flox/flox 小鼠)具有增加的SIRT1活性或24小时禁食的小鼠 (Sir2d小鼠)。喂养高脂饮食 (HFD)的小鼠，并测量血浆胰高血糖素样肽 1 (GLP-1) 和葡萄糖水平。使用定量PCR、免疫印迹和免疫组织化学分析肠道组织、EEC和形成的器官样体。在HFD饲养的VilKO和NgnKO小鼠中，观察到EEC的增加(42.3%和37.2%)，GLP-1或-2产生的L细胞(93.0%和61.4%)，以及GLP-1 (85.7%和109.6%)在葡萄糖负荷后，这解释了HFD-VilKO小鼠代谢表型的改善。这些增加与在HFD-VilKO和HFD-NgnKO小鼠的隐窝中上调的Ngn3 (EEPC 标志物)的表达相关。相反，Sir2d或24小时禁食的小鼠表现出EEC (21.6%)、L细胞 (41.6%)和增殖祖细胞的减少。SIRT1过表达或抑制介导的祖细胞增殖变化与Wnt/β-catenin活性的变化有关。值得注意的是，Wnt/β-catenin抑制剂在HFD-VilKO小鼠或来自HFD-VilKO和HFD-NgnKO小鼠的器官样体中完全抑制了EEC和L细胞数量的增加。肠道中的SIRT1在EEC中通过调节β-catenin活性调控EEPC循环，并可以控制HFD饲养小鼠中的EEC数量，这是一个之前未知的角色。版权所有©2023年作者。由Elsevier Inc.出版。保留所有权利。

We aimed to investigate how SIRT1, a conserved mammalian NAD+-dependent protein deacetylase, regulates the number of enteroendocrine cells (EECs). EECs benefit metabolism, and their increase could potentially treat type 2 diabetes and obesity.We used mice with specific Sirt1 disruption in the intestinal epithelium (VilKO, villin-Cre+, and Sirt1flox/flox mice) or enteroendocrine progenitor cells (EEPCs) (NgnKO, neurogenin3(ngns)-Cre+, Sirt1flox/flox mice) and mice with increased SIRT1 activity due to overexpression (Sir2d mice) or 24 h-fasting. Mice were fed a high-fat diet (HFD), and blood glucagon-like peptide 1 (GLP-1) and glucose levels were measured. Intestinal tissues, EECs, and formed organoids were analyzed using quantitative PCR, immunoblotting, and immunohistochemistry.In HFD-fed VilKO and NgnKO mice, an increase in EECs (42.3% and 37.2%), GLP-1 or -2-producing L cells (93.0% and 61.4%), and GLP-1 (85.7% and 109.6%) was observed after glucose loading, explaining the improved metabolic phenotype of HFD-VilKO mice. These increases were associated with upregulated expression of Ngn3 (EEPC marker) in crypts of HFD-VilKO and HFD-NgnKO mice, respectively. Conversely, Sir2d or 24 h-fasted mice exhibited a decrease in EECs (21.6%), L cells (41.6%), and proliferative progenitor cells. SIRT1 overexpression- or knockdown-mediated change in progenitor cell proliferation was associated with Wnt/β-catenin activity changes. Notably, Wnt/β-catenin inhibitor completely suppressed EEC and L cell increase in HFD-VilKO mice or organoids from HFD-VilKO and HFD-NgnKO mice.Intestinal SIRT1 in EECs modulates the EEPC cycle by regulating β-catenin activity and can control the number of EECs in HFD-fed mice, which is a previously unknown role.Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.