Activation of nicotinamide adenine dinucleotide phosphate (reduced form) oxidase by advanced glycation end products links oxidative stress to altered retinal vascular endothelial growth factor expression.
Increasing evidence indicates that advanced glycation end products (AGEs) promote retinal alterations through oxidative stress. However, the pathways involved in AGE-induced generation of reactive oxygen species (ROS) in retinal cells are poorly defined. In the present study, we investigated the role of nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH) oxidase in AGE-induced ROS intracellular generation and vascular endothelial growth factor (VEGF) expression in bovine retinal endothelial cells (BRECs). Incubation of BRECs with 100 microg/mL AGEs increased ROS generation and VEGF expression in these cells. Treatment of the cells with the NADPH oxidase inhibitors, apocynin and diphenylene iodonium, inhibited these effects. In retinal endothelial cells exposed to AGEs, translocation of protein kinase C (PKC)-beta2 and p47phox was observed. Inhibition of PKC by treatment of the cells with calphostin C, GF10923X, and LY379196 totally suppressed AGE-mediated p47phox translocation and ROS generation. Incubation of BRECs with gliclazide inhibited AGE-induced PKC-beta2 and p47phox translocation and totally abrogated AGE-mediated ROS generation and VEGF expression. Overall, these results demonstrate that AGEs induce intracellular ROS generation and VEGF expression in retinal endothelial cells through a PKC-dependent activation of NADPH oxidase. Inhibition of retinal NADPH oxidase expression and ROS generated by this system provides a new potential mechanism by which gliclazide may affect retinal VEGF expression and exert a beneficial effect on diabetic retinopathy.
Li
Ling
L
CHUM Research Centre, Vascular Immunology Laboratory, Notre-Dame Hospital, Department of Medicine, University of Montreal, Quebec, Canada H2L 4M1.
Renier
Geneviève
G
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D003930:Diabetic Retinopathy / Q000188:drug therapy / Q000201:enzymology / Q000378:metabolism* / Q000517:prevention & control; D042783:Endothelial Cells / Q000201:enzymology / Q000378:metabolism; D004789:Enzyme Activation; D005907:Gliclazide / Q000494:pharmacology; D017127:Glycation End Products, Advanced / Q000378:metabolism*; D007004:Hypoglycemic Agents / Q000494:pharmacology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009861:Onium Compounds / Q000494:pharmacology; D018384:Oxidative Stress / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D064546:Protein Kinase C beta; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012160:Retina / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D000098:Acetophenones; D017127:Glycation End Products, Advanced; D007004:Hypoglycemic Agents; D009861:Onium Compounds; D047428:Protein Kinase Inhibitors; D017382:Reactive Oxygen Species; D042461:Vascular Endothelial Growth Factor A; C007517:diphenyleneiodonium; C056165:acetovanillone; D019255:NADPH Oxidases; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D005907:Gliclazide