Effect of Methylglyoxal on the Oxidative Stress in Trabecular Meshwork Cells

Seung Hee Lee; Sin Hoo Kim; Jae Woo Kim

doi:10.3341/jkos.2009.50.10.1569

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Lee, Kim, and Kim: Effect of Methylglyoxal on the Oxidative Stress in Trabecular Meshwork Cells

Original Article

J Korean Ophthalmol Soc 2009;50(10):1569-1575.

Published online: 3 August 2009

DOI: https://doi.org/10.3341/jkos.2009.50.10.1569

Effect of Methylglyoxal on the Oxidative Stress in Trabecular Meshwork Cells

Seung Hee Lee, MD, Sin Hoo Kim, MD, Jae Woo Kim, MD, PhD

Department of Ophthalmology, Catholic University of Daegu College 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 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

Received 9 April 2009 Accepted 30 June 2009

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 methylglyoxal (MG), intermediate metabolite of advanced glycation end products(AGE), on the induction of oxidative stress in human trabecular meshwork cells (HTMC).

Methods

Primarily cultured HTMC were exposed to at concentrations of 0, 30, 100, and 300 μm of MG for 18 hours, with or without co-exposure to N-acetyl-cysteine. Cellular survival and apoptosis were assessed by MTT assay and flow cytometry using annexin-PI double staining. Production of nitric oxide (NO), superoxide, and reactive oxygen species (ROS) was assessed by Griess assay, cytochrome c assay, and dichlorofluorescein diacetate assay, respectively.

Results

MG did not affect cellular survival at concentrations under 100 μm, but induced apoptosis of HTMC at concentrations over 100 μm. MG decreased NO production, accompanied with increased superoxide production. In addition, MG increased ROS, which were abolished by N-acetylcysteine.

Conclusions

MG induced oxidative stress by decreasing NO production, accompanied by increasing superoxide and ROS productions in HTMC. AGE could induce trabecular meshwork dysfunction.

Keywords: Advanced glycation end products, Methylglyoxal, Oxidative stress, Trabecular meshwork cells

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Figure 1.

Effect of methylglyoxal (MG) on the survival of trabecular meshwork cells. MG and co-exposure of 50 μM N-acetyl-cysteine (NAC) did not affect on the cellular survival significantly except at 300 μM MG. (^* p<0.05)

Figure 2.

Effect of methylglyoxal on the induction of apoptosis in cultured trabecular meshwork cells. Methylglyoxal induced apoptosis from 100 μM. (^* p<0.05)

Figure 3.

Effect of methylglyoxal (MG) on the generation of nitric oxide in cultured trabecular meshwork cells. MG decreased nitric oxide production from 30 μM. (^* p<0.05)

Figure 4.

Effect of methylglyoxal on the generation of superoxide. Methylglyoxal increased superoxide production from 30 μM. (^* p<0.05)

Figure 5.

Effect of methylglyoxal (MG) on the generation of reactive oxygen species. MG increased reactive oxygen species, which were abolished by co-exposed N-acetyl cysteine (NAC). (^*,** p<0.05)

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