JKOS Journal of the Korean Ophthalmological Society J Korean Ophthalmol Soc 0378-6471 2092-9374 Korean Ophthalmological Society 10.3341/jkos.2010.51.2.276 jkos-51-276 Original Article Effect of Glucose on the Production of Reactive Oxygen Species in Retinal Pigment Epithelial Cells LeeJae Hyung MD KimJae Woo MD, PhD Department of Ophthalmology, College of Medicine, Catholic University of Daegu, Daegu, Korea Address reprint requests to Jae Woo Kim, MD, PhD Department of Ophthalmology, College of Medicine, Catholic University of Daegu #3056-6 Daemyeung-4-dong, Nam-gu, Daegu 705-718, Korea Tel: 82-53-650-4728, Fax: 82-53-627-0133, E-mail: jwkim@cu.ac.kr 09 2010 07 09 2010 51 2 276 281 10062009 13102009 Copyright © 2010 Korean Ophthalmological Society 2010 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Purpose

To investigate the effect of high glucose (HG) on the production of reactive oxygen species (ROS) in retinal pigment epithelial (RPE) cells.

 Methods

ARPE-19 cells were exposed to low glucose (5 mM) and high glucose (HG, 25 mM) for three days. Additionally, 50 μ M N-acetyl cysteine (NAC) and 100 μ M L-arginine were co-exposed. Productions of nitric oxide (NO), ROS, and superoxide were determined by Griess assay, DCFH-DA assay, and modified cytochrome c assay, respectively.

 Results

HG increased production of NO, ROS, and superoxide. HG-induced cells increased production of superoxide and ROS but were suppressed by NAC and L-arginine (substrate for NO production) as an antioxidant.

 Conclusions

HG increased ROS production in RPE cells. Thus, HG may cause cellular dysfunction and damage by inducing oxidative stress in RPE cells.

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Effect of high glucose on the survival of ARPE-19 cells. High glucose (HG, 25 mM) increased cellular survival significantly compared with low glucose (LG, 5 mM) (* p<0.05). Co-exposed 50 μM N-acetyl cysteine (NAC) or 100 μM L-arginine (L-Arg) did not affect the survival(p>0.05).

Effect of high glucose on the production of nitric oxide in ARPE-19 cells. High glucose (HG, 25 mM) increased nitric oxide production significantly compared with low glucose (LG, 5 mM) (* p<0.05). Co-exposed 50 μM N-acetyl cysteine (NAC) or 100 μM L-arginine (L-Arg) did not affect the production of nitric oxide compared to HG alone (p>0.05).

Effect of high glucose on the production of superoxide in ARPE-19 cells. High glucose (HG, 25 mM) increased superoxide production significantly compared with low glucose (LG, 5 mM) (* p<0.05). HG-induced increased production of superoxide was abolished by co-exposed 50 μM N-acetyl cysteine (NAC) or 100 μM L-arginine (L-Arg), respectively (** p<0.05).

Effect of high glucose on the production of reactive oxygen species (ROS) in ARPE-19 cells. High glucose (HG, 25 mM) increased ROS production significantly compared with low glucose (LG, 5 mM) (* p<0.05). HG-induced increased production of ROS was abolished by co-exposed 50 μM N-acetyl cysteine (NAC) or 100 μM L-arginine (L-Arg), respectively(** p<0.05).