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Am J Physiol Endocrinol Metab 280: E685-E694, 2001;
0193-1849/01 $5.00
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Vol. 280, Issue 5, E685-E694, May 2001

Activation of NADPH oxidase by AGE links oxidant stress to altered gene expression via RAGE

Marie-Paule Wautier1,2, Olivier Chappey1, Stefano Corda3, David M. Stern4, Ann Marie Schmidt4, and Jean-Luc Wautier1

1 Laboratoire de Biologie Vasculaire et Cellulaire, Hôpital Lariboisière, 2 Unite Institut National de la Santé et de la Recherche Médicale 76, and 3 Laboratoire Microcirculation, Biophysique Hôpital Lariboisière, Paris, France 75475; and 4 College of Physicians and Surgeons, Columbia University, New York, New York 10032

Engagement of the receptor for advanced glycation end products (RAGE) by products of nonenzymatic glycation/oxidation triggers the generation of reactive oxygen species (ROS), thereby altering gene expression. Because dissection of the precise events by which ROS are generated via RAGE is relevant to the pathogenesis of complications in AGE-related disorders, such as diabetes and renal failure, we tested the hypothesis that activation of NADPH oxidase contributed, at least in part, to enhancing oxidant stress via RAGE. Here we show that incubation of human endothelial cells with AGEs on the surface of diabetic red blood cells, or specific AGEs, (carboxymethyl)lysine (CML)-modified adducts, prompted intracellular generation of hydrogen peroxide, cell surface expression of vascular cell adhesion molecule-1, and generation of tissue factor in a manner suppressed by treatment with diphenyliodonium, but not by inhibitors of nitric oxide. Consistent with an important role for NADPH oxidase, although macrophages derived from wild-type mice expressed enhanced levels of tissue factor upon stimulation with AGE, macrophages derived from mice deficient in a central subunit of NADPH oxidase, gp91phox, failed to display enhanced tissue factor in the presence of AGE. These findings underscore a central role of NADPH oxidase in AGE-RAGE-mediated generation of ROS and provide a mechanism for altered gene expression in AGE-related disorders.

diabetes; renal failure; glycation; receptors; cellular activation


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