A Study of the Pathway of Nitric Oxide Production by Nitroglycerin in Trabecular Meshwork Cells

Suk Hyun Yoon; Jae Woo Kim

doi:10.3341/jkos.2013.54.9.1429

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Yoon and Kim: A Study of the Pathway of Nitric Oxide Production by Nitroglycerin in Trabecular Meshwork Cells

Original Article

J Korean Ophthalmol Soc 2013;54(9):1429-1434.

Published online: 7 September 2013

DOI: https://doi.org/10.3341/jkos.2013.54.9.1429

A Study of the Pathway of Nitric Oxide Production by Nitroglycerin in Trabecular Meshwork Cells

Suk Hyun Yoon, 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

∗ Supported by the 2012 Cheil-nammyung Foundation Research Fund.

Received 18 March 201310 April 2013 Accepted 26 July 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 effects of nitroglycerin on the production of nitric oxide and its related pathway in cultured hu-man trabecular meshwork cells (HTMC).

Methods

Primarily cultured HTMC were exposed to 10 nM nitroglycerin using 1% serum-containing media for 30 minutes. The production of nitric oxide was assessed with the Griess assay and expressions of eNOS mRNA was assessed with RT-PCR. Additionally, the cells were exposed to wortmanin and Akt1/2 kinase inhibitor to investigate the mechanism re-lated to the production of nitric oxide.

Results

Nitroglycerin increased the production of nitric oxide (p < 0.05) accompanied with increased expression of eNOS mRNA. The increased production of nitric oxide and eNOS mRNA was inhibited by wortmanin and Akt1/2 kinase inhibitor. Conclusions: Low-dose nitroglycerin increased the production of nitric oxide accompanied by increased eNOS activity. Nitroglycerin drives eNOS activation via the phosphatidylinositol 3-kinase/protein kinase B pathway.

Keywords: eNOS, Nitroglycerin, Nitric oxide, Trabecular meshwork cells

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Figure 1.

Effects of Nitroglycerin (GTN) and vascular endo-thelial growth factor (VEGF) on the production of nitric oxide in trabecular meshwork cells. Both GTN and VEGF increased production of nitric oxide significantly (^* p < 0.05).

Figure 2.

Effects of wortmanin and Akt1/2 kinase inhibitor on the nitroglycerin (GTN)-induced production of nitric oxide in trabecular meshwork cells. Akt1/2 kinase inhibitor and wort-manin inhibits GTN-induced production of nitric oxide (^* p < 0.05).

Figure 3.

Effects of nitroglycerin (GTN) on the activity of eNOS in trabecular meshwork cells. GTN and vascular growth factor (VEGF) increased eNOS expression, which were abolished by Akt1/2 kinase inhibitor and wortmanin compared to the non-exposed control.

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