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

Hyun Yeon Kim; Jae Woo Kim

doi:10.7469/JKOS.2015.56.05.771

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Kim and Kim: Effect of Nitric Oxide on the Permeability of Trabecular Meshwork Cell Monolayer

Original Article

J Korean Ophthalmol Soc 2015;56(5):771-775.

Published online: 7 September 2015

DOI: https://doi.org/10.7469/JKOS.2015.56.05.771

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

Hyun Yeon Kim, MD, Jae Woo Kim, MD, PhD

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 Duryugongwon-ro 17-gil, Nam-gu, Daegu 705-718, Korea Tel: 82-53-650-4728, Fax: 82-53-627-0133 E-mail: jwkim@cu.ac.kr

Received 27 December 20146 January 2015 Accepted 20 April 2015

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.

초록

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 μ m S-nitro-so-N-acetyl-DL-penicillamine (SNAP) and 0.5 mm L-N ^G-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 μ m SNAP ( p > 0.05) but NO production increased in a dose-depend-ent 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 mess-work (TM) relaxation.

Keywords: Carboxyfluorescein, Nitric oxide, Permeability, Trabecular meshwork cells

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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.

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