Effect of High Glucose on the Production of Reactive Oxygen Species in R28 Cells

Jong Min Lee; Jae Woo Kim

doi:10.3341/jkos.2010.51.5.746

Journal List > J Korean Ophthalmol Soc > v.51(5) > 1008819

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Lee and Kim: Effect of High Glucose on the Production of Reactive Oxygen Species in R28 Cells

Original Article

J Korean Ophthalmol Soc 2010;51(5):746-750.

Published online: 25 September 2010

DOI: https://doi.org/10.3341/jkos.2010.51.5.746

Effect of High Glucose on the Production of Reactive Oxygen Species in R28 Cells

Jong Min Lee, MD, Jae Woo Kim, MD, PhD

Department of Ophthalmology, Catholic University of Daegu School of Medicine, Daegu, Korea

Address reprint requests to Jae Woo Kim, MD, PhD Department of Ophthalmology, Catholic University of Daegu College of Medicine #3056-6 Daemyeung4-dong, Nam-gu, Daegu 705-718, Korea Tel: 82-53-650-4728, Fax: 82-53-627-0133, E-mail: jwkim@cu.ac.kr

Received 4 June 2009 Accepted 17 February 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 precursor R28 cells.

Methods

R28 cells were incubated with low glucose (5 mM) or HG (25 mM) for two days. Additionally, the cells were co-exposed to 50 μ M N-acetyl cysteine or 100 μ M L-arginine. Production of nitric oxide (NO), ROS, and superoxide were assessed by Griess assay, DCFH-DA assay, and modified cytochrome c assay, respectively.

Results

HG increased the production of NO, ROS, and superoxide, which were abolished by antioxidants NAC and L-arginine (a substrate for NO production).

Conclusions

HG increased ROS production in R28 cells. Thus, HG may cause cellular dysfunction and damage by inducing oxidative stress in retinal ganglion cells.

Keywords: High glucose, Oxidative stress, Reactive oxygen species, Retinal ganglion cells

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Figure 1.

Effect of glucose on the production of nitric oxide in R28 cells. High glucose (25 mM) induced significantly more production of nitric oxide than low glucose (5 mM). (^*p<0.05)

Figure 2.

High glucose (HG, 25 mM) induced more production of superoxide than low glucose (LG, 5 mM) (^* p>0.05). HG-induced increased production of superoxide was abolished significantly by co-exposed N-acetyl cysteine (NAC) or L-arginine (Arg), respectively. (^** p<0.05)

Figure 3.

High glucose (HG, 25 mM) induced more production of reactive oxygen species (ROS) than low glucose (LG, 5 mM) (^* p>0.05). HG-induced increased production of ROS was abolished significantly by co-exposed N-acetyl cysteine (NAC) or L-arginine (Arg), respectively. (^** p<0.05)

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