J Korean Ophthalmol Soc.  2009 Jun;50(6):919-922. 10.3341/jkos.2009.50.6.919.

Effect of Nitric Oxide on the Survival of R28 Cells

• Lee JI
• Kim JW

• KMID: 2212371
• DOI: http://doi.org/10.3341/jkos.2009.50.6.919

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, Nomega-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.

Keyword

Neurotoxicity; Nitric oxide; R28 cells

MeSH Terms

Culture Media, Serum-Free
NG-Nitroarginine Methyl Ester
Nitric Oxide
Vimentin
Culture Media, Serum-Free
NG-Nitroarginine Methyl Ester
Nitric Oxide
Vimentin

Figure

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  Figure 1. Phase-contrast photograph of R28 cells in monolayer culture (×100).

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  Figure 2. R28 cells immunostained with vimentin (×100).

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  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).

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  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).

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  Figure 5. NO production of R28 cells after exposed to SNAP for 1 day. SNAP increased NO production significantly (* p<0.05).

Reference

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