J Korean Ophthalmol Soc.  2020 Apr;61(4):400-406. 10.3341/jkos.2020.61.4.400.

Effect of Hydrogen Sulfide on the Oxidative Stress and Senescence inTrabecular Meshwork Cells

• Park HJ
• Kim YY
• Kim JW

• KMID: 2500105
• DOI: http://doi.org/10.3341/jkos.2020.61.4.400

Abstract

Purpose
To investigate the effects of hydrogen sulfide (H2S) on the senescence and oxidative stress of cultured human trabecular meshwork cells (HTMCs).
Methods
After inducing oxidative stress with 200 μM hydrogen peroxide (H2O2), primary cultured HTMCs were exposed to 0, 50, and 100 μM sodium hydrogen sulfide (NaHS) for 24 hours. Cell survival and senescence were measured using an MTT assay and SA-β-galactosidase staining, respectively. Generation of reactive oxygen species (ROS) and superoxide were measured using the dichlorofluorescein assay and modified cytochrome c assay. Production of nitric oxide (NO) and expression of eNOS mRNA were assessed using the Griess assay and reverse polymerase chain reaction.
Results
Cell survival was not affected by 200 μM H2O2. Exposure to 50 and 100 μM NaHS significantly decreased cellular senescence, compared with the H2O2-exposed control. Exposure to 50 and 100 μM NaHS decreased the generation of ROS, and exposure to 100 μM NaHS decreased the generation of superoxide. Treatment with 100 μM NaHS increased the production of NO and expression of eNOS mRNA, respectively.
Conclusions
NaHS decreased the cellular senescence associated with the decreased generation of free radicals. Thus, H2S could inhibit the senescence of HTMCs by reducing oxidative stress.

Keyword

Hydrogen sulfide; Oxidative stress; Reactive oxygen species; Senescence; Trabecular meshwork