The uremic solute indoxyl sulfate induces oxidative stress in endothelial cells
The uremic solute indoxyl sulfate induces oxidative stress in endothelial cells
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DOI：
10.1111/j.1538-7836.2007.02540.x
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发表时间：
2007-06-01
影响因子：
10.4
通讯作者：
Brunet, P.
中科院分区：
文献类型：
--
作者：
Dou, L.;Jourde-Chiche, N.;Brunet, P.
背景：内皮功能障碍和氧化应激是慢性肾衰竭（CRF）患者关注的问题。这些患者体内保留的尿毒症溶质可能参与了这些过程。值得注意的是，蛋白质结合的尿毒症溶质硫酸吲哚酚在体外诱导内皮功能障碍，并显示出促氧化作用。目的：证明硫酸吲哚酚是体外内皮细胞氧化应激的潜在介质。方法：通过细胞荧光法测量活性氧 (ROS) 的产生，分析促氧化酶 NAD(P)H 氧化酶、黄嘌呤氧化酶和 NO 合酶的参与，以及测量非酶抗氧化剂谷胱甘肽的水平，研究硫酸吲哚酚诱导的人脐静脉内皮细胞 (HUVEC) 氧化应激。结果：我们发现，无论有或没有人血清白蛋白，硫酸吲哚酚都能诱导 HUVEC 中 ROS 的显着产生。然后我们研究了促氧化酶的作用并测量了抗氧化剂谷胱甘肽的水平。 NAD(P)H 氧化酶抑制剂、DPI 和夹竹桃麻素可抑制 ROS 产生，而黄嘌呤氧化酶、NO 合酶和线粒体 ROS 抑制剂则没有效果。有趣的是，硫酸吲哚酚强烈降低了谷胱甘肽的水平，谷胱甘肽是细胞最活跃的抗氧化系统之一。此外，硫酸吲哚酚介导的ROS产生受到抗氧化剂维生素C、维生素E和NAC的抑制。结论：尿毒症溶质硫酸吲哚酚可增强 ROS 产生，增加 NAD(P)H 氧化酶活性，并降低内皮细胞中谷胱甘肽水平。因此，硫酸吲哚酚通过改变内皮细胞中促抗氧化机制之间的平衡来诱导氧化应激。
Background: Endothelial dysfunction and oxidative stress are matters of concern in patients with chronic renal failure (CRF). Uremic solutes retained in these patients could be involved in these processes. Notably, the protein-bound uremic solute indoxyl sulfate induces endothelial dysfunction in vitro, and has shown pro-oxidant effects. Objective: To demonstrate that indoxyl sulfate is a potential mediator of oxidative stress in endothelial cells in vitro. Methods: Indoxyl sulfate-induced oxidative stress in human umbilical vein endothelial cells (HUVEC) was studied by measuring reactive oxygen specie (ROS) production by cytofluorimetry, by analyzing the involvement of the pro-oxidative enzymes NAD(P)H oxidase, xanthine oxidase, and NO synthase, and by measuring the levels of the non-enzymatic antioxidant glutathione. Results: We showed that indoxyl sulfate induced a significant production of ROS in HUVEC, with or without human serum albumin. We then investigated the role of pro-oxidative enzymes and measured the levels of the antioxidant glutathione. The NAD(P)H oxidase inhibitors, DPI, and apocynin, inhibited ROS production, whereas inhibitors of xanthine oxidase, NO synthase, and mitochondrial ROS had no effect. Interestingly, indoxyl sulfate strongly decreased the levels of glutathione, one of the most active antioxidant systems of the cell. In addition, the ROS production mediated by indoxyl sulfate was inhibited by the antioxidants vitamin C, vitamin E, and NAC. Conclusion: The uremic solute indoxyl sulfate enhances ROS production, increases NAD(P)H oxidase activity, and decreases glutathione levels in endothelial cells. Thus, indoxyl sulfate induces oxidative stress by modifying the balance between pro- and antioxidant mechanisms in endothelial cells.