Department of Metabolic Medicine, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita City, Osaka, 565-0871, Japan.
First Department of Medicine, Wakayama Medical University, Wakayama, Japan.
Biochem Biophys Res Commun. 2021 Jun 4;556:134-141. doi: 10.1016/j.bbrc.2021.03.148. Epub 2021 Apr 8.
Oxidative stress is a deteriorating factor for pancreatic β-cells under chronic hyperglycemia in diabetes. However, the molecular mechanism underlying the increase in oxidative stress in β-cells under diabetic conditions remains unclear. We demonstrated previously that the selective alleviation of glucotoxicity ameliorated the downregulation of several β-cell factors, including Cox6a2. Cox6a2 encodes a subunit of the respiratory chain complex IV in mitochondria. In this study, we analyzed the role of Cox6a2 in pancreatic β-cell function and its pathophysiological significance in diabetes mellitus. Cox6a2-knockdown experiments in MIN6-CB4 cells indicated an increased production of reactive oxygen species as detected by CellROX Deep Red reagent using flow cytometry. In systemic Cox6a2-knockout mice, impaired glucose tolerance was observed under a high-fat high-sucrose diet. However, insulin resistance was reduced when compared with control littermates. This indicates a relative insufficiency of β-cell function. To examine the transcriptional regulation of Cox6a2, ATAC-seq with islet DNA was performed and an open-chromatin area within the Cox6a2 enhancer region was detected. Reporter gene analysis using this area revealed that MafA directly regulates Cox6a2 expression. These findings suggest that the decreased expression of Cox6a2 increases the levels of reactive oxygen species and that Mafa is associated with decreased Cox6a2 expression under glucotoxic conditions.
氧化应激是糖尿病慢性高血糖状态下胰腺 β 细胞恶化的一个因素。然而,糖尿病状态下 β 细胞中氧化应激增加的分子机制尚不清楚。我们之前已经证明,选择性减轻糖毒性可以改善几种 β 细胞因子(包括 Cox6a2)的下调。Cox6a2 编码线粒体呼吸链复合物 IV 的一个亚基。在这项研究中,我们分析了 Cox6a2 在胰腺 β 细胞功能中的作用及其在糖尿病中的病理生理意义。MIN6-CB4 细胞中的 Cox6a2 敲低实验表明,通过流式细胞术使用 CellROX Deep Red 试剂检测到活性氧的产生增加。在系统性 Cox6a2 敲除小鼠中,高脂肪高蔗糖饮食下观察到葡萄糖耐量受损。然而,与对照同窝仔相比,胰岛素抵抗降低。这表明β细胞功能相对不足。为了研究 Cox6a2 的转录调控,对胰岛 DNA 进行了 ATAC-seq 分析,并在 Cox6a2 增强子区域检测到开放染色质区域。使用该区域的报告基因分析表明,Mafa 直接调节 Cox6a2 的表达。这些发现表明,Cox6a2 的表达下调会增加活性氧水平,并且在糖毒性条件下,Mafa 与 Cox6a2 表达的下调有关。