J Korean Ophthalmol Soc.  2015 May;56(5):771-775. 10.3341/jkos.2015.56.5.771.

Effect of Nitric Oxide on the Permeability of Trabecular Meshwork Cell Monolayer

• Kim HY
• Kim JW

• KMID: 2121178
• DOI: http://doi.org/10.3341/jkos.2015.56.5.771

Abstract

PURPOSE
To investigate the effects of nitric oxide (NO) on the permeability of cultured human trabecular meshwork cell (HTMC) monolayer.
METHODS
HTMCs were cultured until confluency in the Transwell inner chamber and then exposed to 0, 10 or 100 microm S-nitroso-N-acetyl-DL-penicillamine (SNAP) and 0.5 mm L-NG-Nitroarginine methyl ester (L-NAME) for 24 hours. Permeabilities of carboxyfluorescein through the HTMC monolayer were measured using a spectrofluorometer after 2 hours in the outer chamber. Cellular viabilities and production of NO were assessed using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and Griess assay, respectively.
RESULTS
The cellular survival was not affected by 10 or 100 microm SNAP (p > 0.05) but NO production increased in a dose-dependent manner (p < 0.05). SNAP significantly increased the permeability of carboxyfluorescein through the HTMC monolayer in a dose-dependent manner compared with non-exposed control (p < 0.05). The endothelial NO synthase inhibitor L-NAME abolished SNAP-induced increase of the carboxyfluorescein permeability (p > 0.05).
CONCLUSIONS
NO increased the permeability of carboxyfluorescein through the HTMC monolayer in a dose-dependent manner. Thus, NO could increase trabecular outflow by increasing the permeability of trabecular cell layer in addition to trabeular messwork (TM) relaxation.

Keyword

Carboxyfluorescein; Nitric oxide; Permeability; Trabecular meshwork cells

MeSH Terms

Humans
NG-Nitroarginine Methyl Ester
Nitric Oxide Synthase
Nitric Oxide*
Permeability*
Relaxation
Trabecular Meshwork*
NG-Nitroarginine Methyl Ester
Nitric Oxide
Nitric Oxide Synthase

Figure

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  Figure 1. Effect of nitric oxide donor on the survival of confluently cultured trabecular meshwork cells in monolayer. 10, 100 μ m SNAP and co-exposed 0.5 mm L did not affect on the survival significantly compared to non-exposed control ( p > 0.05). SNAP = S-Nitroso-N-acetyl-DL-penicillamine; L = L-N G- Nitroarginine methyl ester.

• 

  Figure 2. Effect of nitric oxide donor on the production of nitrite in confluently cultured trabecular meshwork cells. 10, 100 μ m SNAP, and 100 μ m SNAP with 0.5 mm L increased nitrite production significantly compared to non-exposed control. SNAP = S-Nitroso-N-acetyl-DL-penicillamine; L = L-N G-Nitroarginine methyl ester. * p < 0.05.

• 

  Figure 3. Effects of nitric oxide donor on the permeability of carboxyfluorescin through the trabecular meshwork cell monolayer. 10, 100 μ m SNAP increased the permeability of carboxyfluorescin significantly compared to non-exposed control and abolished by co-exposed 0.5 mm L. Carboxyfluorescein intensity of outer chamber normalized to the mean value obtained using non-ex-posed control (permeability 100%). SNAP = S-Nitroso-N- ace-tyl-DL-penicillamine; L = L-N G-Nitroarginine methyl ester. * p < 0.05.

Reference

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