Allopurinol protects human glomerular endothelial cells from high glucose-induced reactive oxygen species generation, p53 overexpression and endothelial dysfunction
Allopurinol protects human glomerular endothelial cells from high glucose-induced reactive oxygen species generation, p53 overexpression and endothelial dysfunction
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DOI：
10.1007/s11255-017-1733-5
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发表时间：
2018-01-01
影响因子：
2
通讯作者：
Stefanidis, Ioannis
中科院分区：
文献类型：
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作者：
Eleftheriadis, Theodoros;Pissas, Georgios;Stefanidis, Ioannis
毛细血管内皮细胞线粒体活性氧（ROS）过量产生是糖尿病肾病发生的先决条件。黄嘌呤氧化酶（另一种 ROS 生成剂）的抑制可改善实验性糖尿病肾病。为了验证线粒体最初高葡萄糖诱导的 ROS 产生会激活黄嘌呤氧化酶（此后黄嘌呤氧化酶仍然是 ROS 的主要来源）的假设，我们在正常或高葡萄糖条件下培养原代人肾小球内皮细胞 (GEnC)，有或没有黄嘌呤氧化酶抑制剂别嘌呤醇。通过化学发光或比色法评估 ROS 的产生和一氧化氮合酶 (NOS) 活性。通过蛋白质印迹法评估细胞间粘附分子 1 (ICAM-1)、p53 和磷酸化 p53 (p-p53) 的水平。别嘌呤醇可防止高葡萄糖诱导的 ROS 生成，表明黄嘌呤氧化酶是 ROS 的主要来源。别嘌呤醇可以保护 GEnC 免受内皮功能障碍的影响，因为它可以防止高葡萄糖诱导的 NOS 活性下降和 ICAM-1 表达增加。别嘌呤醇降低高糖诱导的 p53 和 p-p53 水平，表明黄嘌呤氧化酶衍生的 ROS 轴、DNA 损伤、p53 稳定和内皮功能障碍可能导致糖尿病肾病的发病机制。别嘌呤醇可保护 GEnC 免受高糖诱导的 ROS 生成、p53 过度表达和内皮功能障碍的影响。这些数据提供了一种发病机制，支持黄嘌呤氧化酶抑制剂对糖尿病肾病发展的有益作用的实验和临床研究结果。
Mitochondrial reactive oxygen species (ROS) overproduction in capillary endothelial cells is a prerequisite for the development of diabetic nephropathy. Inhibition of xanthine oxidase, another ROS generator, ameliorates experimental diabetic nephropathy. To test the hypothesis that the initial high glucose-induced ROS production by the mitochondria activates xanthine oxidase, which afterward remains as the major source of ROS, we cultured primary human glomerular endothelial cells (GEnC) under normal or high-glucose conditions, with or without the xanthine oxidase inhibitor allopurinol.ROS generation and nitric oxide synthase (NOS) activity were assessed by chemiluminescence or colorimetrically. Levels of intercellular adhesion molecule 1 (ICAM-1), p53 and phosphorylated p53 (p-p53) were assessed by western blotting.Allopurinol prevented high glucose-induced ROS generation indicating that xanthine oxidase is the major source of ROS. Allopurinol protected GEnC from endothelial dysfunction since it prevented the high glucose-induced decrease in NOS activity and increase in ICAM-1 expression. Allopurinol reduced p53 and p-p53 levels induced by high glucose suggesting an axis of xanthine oxidase-derived ROS, DNA damage, p53 stabilization and endothelial dysfunction that may contribute to the pathogenesis of diabetic nephropathy.Allopurinol protects GEnC from high glucose-induced ROS generation, p53 overexpression and endothelial dysfunction. These data provide a pathogenetic mechanism that supports the results of experimental and clinical studies about the beneficial effect of xanthine oxidase inhibitors on the development of diabetic nephropathy.