School of Pharmacy, University of Waterloo, Kitchener, ON, Canada.
Institute of Metabolism and Systems Research (IMSR), University of Birmingham, Birmingham, UK.
Endocrinology. 2022 Jan 1;163(1). doi: 10.1210/endocr/bqab226.
Pancreatic β-cells can secrete insulin via 2 pathways characterized as KATP channel -dependent and -independent. The KATP channel-independent pathway is characterized by a rise in several potential metabolic signaling molecules, including the NADPH/NADP+ ratio and α-ketoglutarate (αKG). Prolyl hydroxylases (PHDs), which belong to the αKG-dependent dioxygenase superfamily, are known to regulate the stability of hypoxia-inducible factor α. In the current study, we assess the role of PHDs in vivo using the pharmacological inhibitor dimethyloxalylglycine (DMOG) and generated β-cell-specific knockout (KO) mice for all 3 isoforms of PHD (β-PHD1 KO, β-PHD2 KO, and β-PHD3 KO mice). DMOG inhibited in vivo insulin secretion in response to glucose challenge and inhibited the first phase of insulin secretion but enhanced the second phase of insulin secretion in isolated islets. None of the β-PHD KO mice showed any significant in vivo defects associated with glucose tolerance and insulin resistance except for β-PHD2 KO mice which had significantly increased plasma insulin during a glucose challenge. Islets from both β-PHD1 KO and β-PHD3 KO had elevated β-cell apoptosis and reduced β-cell mass. Isolated islets from β-PHD1 KO and β-PHD3 KO had impaired glucose-stimulated insulin secretion and glucose-stimulated increases in the ATP/ADP and NADPH/NADP+ ratio. All 3 PHD isoforms are expressed in β-cells, with PHD3 showing the most distinct expression pattern. The lack of each PHD protein did not significantly impair in vivo glucose homeostasis. However, β-PHD1 KO and β-PHD3 KO mice had defective β-cell mass and islet insulin secretion, suggesting that these mice may be predisposed to developing diabetes.
胰岛β细胞可以通过两种途径分泌胰岛素,这两种途径的特征分别为依赖和不依赖 KATP 通道。不依赖 KATP 通道的途径表现为几种潜在代谢信号分子的增加,包括 NADPH/NADP+ 比和α-酮戊二酸(αKG)。脯氨酰羟化酶(PHD)属于 αKG 依赖性双加氧酶超家族,已知其可以调节缺氧诱导因子α的稳定性。在本研究中,我们使用药理学抑制剂二甲基草酰甘氨酸(DMOG)和生成β细胞特异性敲除(KO)小鼠来评估 PHD 在体内的作用,这些 KO 小鼠敲除了所有 3 种 PHD 同工型(β-PHD1 KO、β-PHD2 KO 和β-PHD3 KO 小鼠)。DMOG 抑制了体内葡萄糖刺激的胰岛素分泌,并抑制了胰岛素分泌的第一阶段,但增强了分离胰岛的第二阶段胰岛素分泌。除了β-PHD2 KO 小鼠在葡萄糖挑战期间胰岛素水平显著升高外,所有β-PHD KO 小鼠均未显示出任何与葡萄糖耐量和胰岛素抵抗相关的明显体内缺陷。β-PHD1 KO 和β-PHD3 KO 的胰岛β细胞凋亡增加,β细胞质量减少。β-PHD1 KO 和β-PHD3 KO 的分离胰岛葡萄糖刺激的胰岛素分泌受损,葡萄糖刺激的 ATP/ADP 和 NADPH/NADP+ 比值增加。所有 3 种 PHD 同工型都在β细胞中表达,其中 PHD3 表现出最明显的表达模式。缺乏每种 PHD 蛋白并没有显著损害体内葡萄糖稳态。然而,β-PHD1 KO 和β-PHD3 KO 小鼠的β细胞质量和胰岛胰岛素分泌受损,表明这些小鼠可能易患糖尿病。