J Korean Ophthalmol Soc.  2020 Apr;61(4):394-399. 10.3341/jkos.2020.61.4.394.

Cytoprotective Effect of Rho Kinase Inhibitors against Oxidative Stress inTrabecular Meshwork Cells

• Kim JW
• Son MS
• Baek MJ

• KMID: 2500104
• DOI: http://doi.org/10.3341/jkos.2020.61.4.394

Abstract

Purpose
We evaluated the protective effect of trabecular outflow drugs, Rho kinase (ROCK) inhibitors against oxidative stress in trabecular meshwork cells.
Methods
Primary-cultured human trabecular meshwork cells (HTMCs) were exposed to ROCK inhibitors at 10 and 20 μM Y-27632, ripasudil or fasudil for 24 hours, after pretreatment with 200 μM hydrogen peroxide for 30 min. The cell viabilities and metabolic activities were assessed using the Trypan Blue dye exclusion test and MTT assay, respectively. Reactive oxygen species (ROS) production was measured using the H2DCFDA assay, and the degree of apoptosis was measured with flow cytometry using annexin-propidium iodide double staining.
Results
In HTMCs, Y-27632 suppressed ROS production. Ripasudil and fasudil increased the metabolic activities and decreased the degrees of apoptosis. Fasudil showed the most cytoprotective effects among the three ROCK inhibitors tested.
Conclusions
Against oxidative stress, ROCK inhibitors decreased apoptosis accompanied by decreased ROS production, especially fasudil. ROCK inhibitors may therefore have cytoprotective properties in addition to increasing trabecular outflow.

Keyword

Apoptosis; Oxidative stress; Rho-associated kinase inhibitor; Trabecular meshwork