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Lee and Kim: Effect of Nitric Oxide on the Survival of R28 Cells
Original Article

J Korean Ophthalmol Soc 2009;50(6):919-922.

Published online: 7 September 2009

DOI: https://doi.org/10.3341/jkos.2009.50.6.919

Effect of Nitric Oxide on the Survival of R28 Cells

Jeong Il Lee, 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 Opthalmology, 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 11 December 2008       Accepted 10 March 2009

Copyright © 2009 Korean Ophthalmological Society

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 evaluate the effect of nitric oxide (NO) on the survival of R28 cells.

Methods

After immunostaining for GFAP, vimentin, S-100, and neurofilament, R28 cells were exposed to S-nitroso-N-acetyl-D, L-penicillamine (SNAP) at various concentrations, with and without the NO inhibitor, Nω-Nitro-L-arginine methyl ester (L-NAME) for 1 and 3 days. Cellular survival of R28 cells and the production of NO were quantified by rapid colorimetric assays using the MTT and Griess assay, respectively. To evaluate the effect of serum, 10% serum or serum-free media were used separately.

Results

R28 cells showed strong immunoreactivity to GFAP and vimentin compared to S-100 or neurofilament. SNAP inhibited the survival of R28 cells in a dose-dependent manner, and this effect of NO on the cellular survival was abolished by L-NAME. These results were similar after exposure for 1 and 3 days, regardless of the presence of serum in the media.

Conclusions

The current results suggest that NO decreased the survival of R28 cells. Further studies are necessary to evaluate the mechanism of cytotoxicity of the R28 cells.

Keywords: Neurotoxicity, Nitric oxide, R28 cells

References

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Figure 1.
Phase-contrast photograph of R28 cells in monolayer culture (×100).
jkos-50-919f1.tif
Figure 2.
R28 cells immunostained with vimentin (×100).
jkos-50-919f2.tif
Figure 3.
Survival of R28 cells after exposed to SNAP for 1 day in serum-free and 10% serum-containing media. Cellular survival was decreased significantly with exposure to NO in a dose-dependent manner (* p<0.05).
jkos-50-919f3.tif
Figure 4.
Survival of R28 cells after exposed to SNAP for 3 days in serum-free and 10% serum-containing media. Cellular survival was decreased significantly with exposure to NO in a dose-dependent manner (* p<0.05).
jkos-50-919f4.tif
Figure 5.
NO production of R28 cells after exposed to SNAP for 1 day. SNAP increased NO production significantly (* p<0.05).
jkos-50-919f5.tif
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