Metformin induces suppression of NAD(P)H oxidase activity in podocytes
Metformin induces suppression of NAD(P)H oxidase activity in podocytes
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DOI：
10.1016/j.bbrc.2010.01.119
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发表时间：
2010-03-05
影响因子：
3.1
通讯作者：
Angielski, Stefan
中科院分区：
文献类型：
--
作者：
Piwkowska, Agnieszka;Rogacka, Dorota;Angielski, Stefan
高血压增加了活性氧（ROS）的产生。NAD（P）H氧化酶是糖尿病足细胞内ROS的主要来源，产生超氧阴离子，参与糖尿病肾病的发生发展。我们研究了抗糖尿病药物二甲双胍对培养足细胞产生超氧阴离子的影响，并试图阐明其潜在的机制。通过DCF探针的荧光测量总体ROS产生。化学发光法测定NAD（P）H氧化酶活性。AMP依赖性激酶（AMPK）活性通过免疫印迹测定，测量磷酸化AMPK与总AMPK的比率。使用2-脱氧-[1，2-（3）H]-葡萄糖测量葡萄糖积累。HG中的活性氧产生增加了约27%（187 +/- 8 vs. 238 +/- 9任意单位Au，P < 0.01）。二甲双胍（2 mM，2小时）显着减少ROS的产生45%，在NG和60%，在HG。二甲双胍使NG和HG的NAD（P）H氧化酶活性分别降低36%和86%。二甲双胍可增加NG和HIS中AMPK活性（从0.58 +/- 0.07增加到0.58 +/- 0.07）。0.99+/- 0.06和0.53 +/- 0.03至0.64 +/- 0.03; P < 0.05）。二甲双胍对NAD（P）H氧化酶和AMPK活性的影响在AMPK抑制剂化合物C存在下被消除。我们已经表明二甲双胍通过降低NAD（P）H氧化酶活性来降低ROS的产生。我们还证实了NAD（P）H氧化酶活性与AMPK活性的关系。(C)2010年爱思唯尔公司All rights reserved.
Hyperglycemia increases the production of reactive oxygen species (ROS). NAD(P)H oxidase, producing superoxide anion, is the main source of ROS in diabetic podocytes and their production contributes to the development of diabetic nephropathy. We have investigated the effect of an antidiabetic drug, metformin on the production of superoxide anion in cultured podocytes and attempted to elucidate underlying mechanisms.The experiments were performed in normal (NG, 5.6 mM) and high (HG, 30 mM) glucose concentration. Overall ROS production was measured by fluorescence of a DCF probe. Activity of NAD(P)H oxidase was measured by chemiluminescence method. The AMP-dependent kinase (AMPK) activity was determined by immunobloting, measuring the ratio of phosphorylated AMPK to total AMPK. Glucose accumulation was measured using 2-deoxy-[ 1,2-(3)H]-glucose.ROS production increased by about 27% (187 +/- 8 vs. 238 +/- 9 arbitrary units AU, P < 0.01) in HG. Metformin (2 mM, 2 h) markedly reduced ROS production by 45% in NG and 60% in HG. Metformin decreased NAD(P)H oxidase activity in NG (36%) and HG (86%). AMPK activity was increased by metformin in NG and HIS (from 0.58 +/- 0.07 to. 0.99 +/- 0.06, and from 0.53 +/- 0.03 to 0.64 +/- 0.03; P < 0.05). The effects of metformin on the activities of NAD(P)H oxidase and AMPK were abolished in the presence of AMPK inhibitor, compound C.We have shown that metformin decreases production of ROS through reduction of NAD(P)H oxidase activity. We also have demonstrated relationship between activity of NAD(P)H oxidase and AMPK. (C) 2010 Elsevier Inc. All rights reserved.