Effect of Ascorbic Acid Against the Oxidative Stress-Induced Cellular Senescence in Trabecular Meshwork Cells

Jae Woo Kim; Sun Hee Kang; Keun Woo Lee

doi:10.3341/jkos.2013.54.3.490

Journal List > J Korean Ophthalmol Soc > v.54(3) > 1009626

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Kim, Kang, and Lee: Effect of Ascorbic Acid Against the Oxidative Stress-Induced Cellular Senescence in Trabecular Meshwork Cells

Original Article

J Korean Ophthalmol Soc 2007;54(3):490-495.

Published online: 24 August 2007

DOI: https://doi.org/10.3341/jkos.2013.54.3.490

Effect of Ascorbic Acid Against the Oxidative Stress-Induced Cellular Senescence in Trabecular Meshwork Cells

Jae Woo Kim, MD, PhD, Sun Hee Kang, MD, Keun Woo Lee, MD

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

Address reprint requests to Jae Woo Kim, MD, PhD Department of Ophthalmology, Daegu Catholic University Medical Center #33 17-gil Duryugongwon-ro, Nam-gu, Daegu 705-718, Korea Tel: 82-53-650-4728, Fax: 82-53-627-0133, E-mail: jwkim@cu.ac.kr

Received 6 April 201220 April 2012 Accepted 16 January 2013

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 L-ascorbic acid (LAA) on oxidative-stress-induced cellular senescence in trabecular meshwork (TM) cells.

Methods

Primarily cultured human TM cells were exposed to 0, 10, or 100 μM hydrogen peroxide for 7 days with or without co-exposure of LAA. Cellular survival and nitrite production were assessed with MTT and Griess assays, respectively. SA-β-gal staining was performed to quantify cellular senescence.

Results

Hydrogen peroxide decreased cellular survival, accompanied by decreased nitric oxide (NO) production. These decreases of cellular survival and NO production were abolished by co-exposure of 100 μM LAA. Analysis of SA-β-gal staining revealed that LAA inhibited hydrogen peroxide-induced cellular senescence by 6.8% (p < 0.05).

Conclusions

LAA may have a protective effect against the oxidative-stress-induced cellular senescence in TM cells.

Keywords: Ascorbic acid, Oxidative stress, Senescence, Trabecular meshwork cells

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Figure 1.

Effect of hydrogen peroxide on the survival of human trabecular meshwork cells. Exposure to 100 μM hydrogen peroxide decreased cellular proliferation significantly compared to non-exposed control (^*p < 0.05).

Figure 2.

Effect of L-ascorbic acid on the survival of human trabecular meshwork cells against the hydrogen peroxide-induced oxidative stress. 100 μM hydrogen peroxide decreased cellular survival significantly (^*p < 0.05) which were abolished by L-ascorbic acid (p > 0.05).

Figure 3.

Effect of hydrogen peroxide on the production of nitric oxide (NO) in primarily cultured human trabecular meshwork cells. Exposure to 100 μM hydrogen peroxide decreased production of NO significantly compared to non-exposed control (^*p < 0.05).

Figure 4.

Effect of L-ascorbic acid on the production of nitric oxide (NO) in human trabecular meshwork cells against the hydrogen peroxide-induced oxidative stress. L-ascorbic acid increased production of NO compared to non-exposed control (^*p < 0.05).

Figure 5.

Senescence-associated β-galactosidase staining of trabecular meshwork cells after exposure to hydrogen peroxide. 100 μM hydrogen peroxide increased the number of senescent cells, which stained blue (B), compared to non-exposed control (A) (Magnification, ×100).

Figure 6.

Effect of L-ascorbic acid on the oxidative stress-induced senescence of human trabecular meshwork cells. 100 μM L-ascorbic acid decreased cellular senescence (^*p < 0.05).

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