Effect of advanced glycation end products on accelerated apoptosis of retinal capillary cells under in vitro conditions

Advanced glycation end-products (AGEs) are considered to play an important role in the development of retinopathy in diabetes, and are shown to induce retinal vascular changes resembling that of diabetic retinopathy. We have shown that apoptosis of retinal capillary cells is accelerated in diabetes. The aim of this study is to investigate the role of AGEs in accelerated retinal capillary cell death in in vitro conditions, and to identify the possible mechanism involved. Bovine retinal endothelial cells and pericytes were incubated in the presence of 5 muM AGE-bovine serum albumin (AGE-BSA) or untreated control BSA (BSA) for up to five days. The cell death was determined by performing ELISA for cytoplasmic histone-associated DNA fragments and by measuring the activity of caspase-3. Incubation of endothelial cells or pericytes with AGE-BSA increased oxidative stress and NO by 60%, and in the same cells nuclear transcriptional factor (NF-kB) was also activated by over 60%. AGE-BSA induced their apoptosis by 55%, and activated caspase-3 by about 50% compared to the cells incubated with unmodified BSA. Co-addition of AGE-BSA and antioxidants (N-acetyl cysteine or alpha-lipoic acid) inhibited oxidative stress, nitrotyrosine formation, NF-kB activation and capillary cell apoptosis. These data strongly suggest that increased AGE in diabetes could play an important role in retinal capillary apoptosis and that oxidative stress is involved in this process. Inhibition of AGEs in the retinal capillary cells could prevent their apoptosis, and ultimately, the development of retinopathy in diabetes. (C) 2004 Elsevier Inc. All rights reserved.