Effects of Hydrogen Sulfide and Nitric Oxide on the Permeability of Cultured Trabecular Meshwork Cells

Myung Seo Son; Min Ju Baek; Jae Woo Kim

doi:10.3341/jkos.2020.61.3.267

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Son, Baek, and Kim: Effects of Hydrogen Sulfide and Nitric Oxide on the Permeability of Cultured Trabecular Meshwork Cells

Original Article

J Korean Ophthalmol Soc 2020;61(3):267-273.

Published online: 25 September 2020

DOI: https://doi.org/10.3341/jkos.2020.61.3.267

Effects of Hydrogen Sulfide and Nitric Oxide on the Permeability of Cultured Trabecular Meshwork Cells

Myung Seo Son, MD, Min Ju Baek, MD, Jae Woo Kim, MD, PhD

Department of Ophthalmology, Daegu Catholic University College of Medicine, Daegu, Korea

Address reprint requests to Jae Woo Kim, MD, PhD Department of Ophthalmology, Daegu Catholic University Hospital, #33 Duryugongwon-ro 17gil, Nam-gu, Daegu 42472, Korea Tel: 82-53-650-4728, Fax: 82-53-627-0133 E-mail: jwkim@cu.ac.kr

Received 4 April 2019 Revised 20 August 2019 Accepted 21 February 2020

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 hydrogen sulfide (H₂ S) on the permeability of a cultured human trabecular meshwork cells (HTMC) monolayer and its interaction with nitric oxide (NO).

Methods

After exposing primary cultured HTMCs to 0, 50, 100, and 500 μ M sodium hydrogen sulfide (NaHS) for 6 hours, the permeabilities through the HTMC monolayer were measured using a Transwell assay with carboxyfluorescein. The production of NO and eNOS mRNA expression were assessed using the Griess assay and reverse transcription-polymerase chain reaction, respectively. In addition, 0, 1, and 10 μ M NaHS and 10 μ M sodium nitroprusside (SN) were co-exposed to evaluate the possible synergistic effect of H₂ S and NO.

Results

Greater than 100 μ M NaHS increased the permeability through the HTMC monolayer in a dose-dependent manner (p < 0.05). These increased permeabilities were not accompanied by NO production or eNOS mRNA expression (p > 0.05). When 0, 1, and 10 μ M NaHS and 10 μ M SN were exposed together, there was no significant change of permeability, NO production, or eNOS mRNA expression (all, p > 0.05).

Conclusions

NaHS at high concentrations increased the permeability of the HTMC monolayer, which was not affected by NO. NaHS at low concentrations did not show a synergistic effect with NO. Thus, H₂ S at high concentrations may increase trabecular outflow, which may not be associated with NO.

Keywords: Hydrogen sulfide, Nitric oxide, Permeability, Trabecular meshwork

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Figure 1.

The Effect of sodium hydrogen sulfide (NaHS; 0, 50,100, 500 μ M) and L-cysteine (100 μ M) on the survival of cultured human trabecular meshwork cells. Both NaHS and L-cys-teine did not affect the survival of trabecular meshwork cells compared to non-exposed controls (all p > 0.05).

Figure 2.

The Effect of sodium hydrogen sulfide (NaHS; 0, 50,100, 500 μ M) and L-cysteine (100 μ M) on the production of nitric oxide in cultured human trabecular meshwork cells. Both NaHS and L-cysteine did not affect the production of nitric oxide compared to non-exposed controls (all p > 0.05).

Figure 3.

The Effect of sodium hydrogen sulfide (NaHS; 0, 50, 100, 500 μ M) and L-cysteine (100 μ M) on the expression of endothelial nitric oxide synthase (eNOS) mRNA in cultured human trabecular meshwork cells. Both NaHS and L-cysteine did not affect the on the expression of eNOS mRNA compared to non-exposed controls (all p > 0.05).

Figure 4.

The Effect of sodium hydrogen sulfide (NaHS: 0, 50,100, 500 μ M) and L-cysteine (100 μ M) on the permeability of carboxyfluorescin through the trabecular meshwork cell monolayer. Both 100, 500 μ M NaHS and 100 μ M L-cysteine increased the permeabilty of carboxyfluorescein significantly (^* p < 0.05). Carboxyfluorescein intensity of outer chamber normalized to the mean value obtained using non-exposed control (permeability 100%).

Figure 5.

The Effect of sodium hydrogen sulfide (NaHS) at low concentration (1, 10 μ M) on the production of nitric oxide in cultured human trabecular meshwork cells. NaHS at low concentration did not affect the production of nitric oxide (all p >0.05).

Figure 6.

Effect of sodium hydrogen sulfide (NaHS) at low concentration (1, 10 μ M) and N_ω-Nitro-L-arginine methyl ester hydrochloride (L-NAME, 10 μ M) on the expression of endothelial nitric oxide synthase (eNOS) mRNA in cultured human trabecular meshwork cells. NaHS did not affect the on the expression of eNOS mRNA compared to non-exposed controls (all p > 0.05). In contrast, L-NAME decreased the expression of eNOS mRNA significantly (p = 0.046).

Figure 7.

Effect of 10 μ M sodium nitroprusside (SN) co-ex-posed to sodium hydrogen sulfide (NaHS) at low concentration (1, 10 μ M) on the permeability of carboxyfluorescein through the trabecular meshwork cell monolayer. Both 1, 10 μ M NaHS co-exposed with SN did not affect the permeabilty of carboxyfluorescein (p = 0.74, p = 0.511) compared with exposed to SN alone. Carboxyfluorescein intensity of outer chamber normalized to the mean value obtained using SN (permeability 100%).

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