Metformin and liraglutide ameliorate high glucose-induced oxidative stress via inhibition of PKC-NAD(P)H oxidase pathway in human aortic endothelial cells
Metformin and liraglutide ameliorate high glucose-induced oxidative stress via inhibition of PKC-NAD(P)H oxidase pathway in human aortic endothelial cells
复制标题
二甲双胍和利拉鲁肽通过抑制人主动脉内皮细胞中的 PKC-NAD(P)H 氧化酶通路改善高血糖诱导的氧化应激反应
DOI：
10.1016/j.atherosclerosis.2013.10.025
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发表时间：
2014-01-01
期刊：
影响因子：
5.3
通讯作者：
Takayanagi, Ryoichi
中科院分区：
文献类型：
--
作者：
Batchuluun, Battsetseg;Inoguchi, Toyoshi;Takayanagi, Ryoichi
目的：二甲双胍和胰高血糖素样肽-1（GLP-1）预防糖尿病心血管并发症和动脉粥样硬化。然而，对高血糖诱导的内皮细胞氧化应激的直接影响还没有完全了解。因此，我们的目的是评估二甲双胍和GLP-1类似物利拉鲁肽对高糖诱导的氧化应激的影响。在接受和不接受二甲双胍和利拉鲁肽治疗的人主动脉内皮细胞中评价了活性氧（ROS）的产生、蛋白激酶C（PKC）和NAD（P）H氧化酶的活化以及信号分子对高糖暴露的反应变化，单独或组合使用。PKC-NAD（P）H氧化酶途径通过GFP融合的PKC β 2亚型和GFP融合的p47 phox（NAD（P）H氧化酶的调节亚基）的移位以及内源性PKC磷酸化和NAD（P）H氧化酶活性来评估。暴露于高糖导致PKC β 2易位和内源性PKC的时间依赖性磷酸化，但在二甲双胍和利拉鲁肽处理的细胞中未能诱导其易位和磷酸化。此外，两种药物均能抑制p47 phox易位和NAD（P）H氧化酶的激活，并阻止高糖诱导的细胞内二酰甘油（DAG）水平和AMP活化蛋白激酶（AMPK）磷酸化的变化。这些药物的组合进一步增强了所有这些effects.Conclusions：美替卡松和利拉鲁肽通过抑制PKC-NAD（P）H氧化酶途径改善高糖诱导的氧化应激。这两种药物的组合提供了增强的保护作用，表明在预防糖尿病血管并发症的临床实用性。(C)2013爱思唯尔爱尔兰有限公司版权所有。
Objective: Metformin and glucagon like peptide-1 (GLP-1) prevent diabetic cardiovascular complications and atherosclerosis. However, the direct effects on hyperglycemia-induced oxidative stress in endothelial cells are not fully understood. Thus, we aimed to evaluate the effects of metformin and a GLP-1 analog, liraglutide on high glucose-induced oxidative stress.Methods: Production of reactive oxygen species (ROS), activation of protein kinase C (PKC) and NAD(P) H oxidase, and changes in signaling molecules in response to high glucose exposure were evaluated in human aortic endothelial cells with and without treatment of metformin and liraglutide, alone or in combination. PKC-NAD(P) H oxidase pathway was assessed by translocation of GFP-fused PKC beta 2 isoform and GFP-fused p47phox, a regulatory subunit of NAD(P) H oxidase, in addition to endogenous PKC phosphorylation and NAD(P) H oxidase activity.Results: High glucose-induced ROS overproduction was blunted by metformin or liraglutide treatment, with a further decrease by a combination of these drugs. Exposure to high glucose caused PKC beta 2 translocation and a time-dependent phosphorylation of endogenous PKC but failed to induce its translocation and phosphorylation in the cells treated with metformin and liraglutide. Furthermore, both drugs inhibited p47phox translocation and NAD(P) H oxidase activation, and prevented the high glucose-induced changes in intracellulalr diacylglycerol (DAG) level and phosphorylation of AMP-activated protein kinase (AMPK). A combination of these drugs further enhanced all of these effects.Conclusions: Metformin and liraglutide ameliorate high glucose-induced oxidative stress by inhibiting PKC-NAD(P) H oxidase pathway. A combination of these two drugs provides augmented protective effects, suggesting the clinical usefulness in prevention of diabetic vascular complications. (C) 2013 Elsevier Ireland Ltd. All rights reserved.