Effect of Chronic Benzalkonium Chloride Exposure on Senescence in Trabecular Meshwork Cells
10.3341/jkos.2019.60.1.55
- Author:
Jihae PARK
1
;
Soo Jin LEE
;
Kyoo Won LEE
;
Jae Chan KIM
Author Information
1. Cheil Eye Research Institute, Daegu, Korea. jck50ey@daum.net
- Publication Type:Original Article
- Keywords:
Benzalkonium chloride;
Senescence;
Trabecular meshwork
- MeSH:
Aging;
Benzalkonium Compounds;
Blotting, Western;
Cell Aging;
Cell Cycle;
Cell Cycle Checkpoints;
Cell Survival;
Cyclins;
Glaucoma;
Rivers;
Trabecular Meshwork
- From:Journal of the Korean Ophthalmological Society
2019;60(1):55-61
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: To determine the possible effects of chronic exposure of low dose benzalkonium chloride (BAK) on trabecular meshwork cells, and to characterize the pathways involved in the effects. METHODS: Trabecular meshwork cells were treated with 0.0005%, 0.00075%, 0.001%, and 0.0025% BAK for 10 minutes; then, the cells were transferred to a new medium for 24 hours. This process was repeated three times. Cell survival was assessed using the MTT assay to determine the non-apoptotic BAK concentration. Senescence-associated (SA)-β-gal staining was performed to compare quantitatively the cellular senescence of BAK-treated cells with the control group. Cells treated with BAK were analyzed by western blot to determine whether the expressions of cell cycle regulators were affected. RESULTS: Two concentrations (0.0005% and 0.00075%) showed persistent cell viability and were chosen for further experiments. After SA-β-gal staining, cells treated with 0.0005% and 0.00075% BAK showed 28% (± 2.08), 37% (± 2.08) increases in cellular senescence expression, respectively, when compared with control cells (p < 0.05). To identify the molecular pathways involved in cell cycle arrest via BAK, western blot analysis was performed on trabecular meshwork cells, resulting in decreased expressions of cyclin E/CDK2, and increased expressions of the upper stream control molecules, p53 and p21. CONCLUSIONS: Chronic exposure to low dose BAK accelerated cell senescence through cell cycle arrest. Because senescent cells of the trabecular meshwork can inhibit its outflow pathway function and ultimately worsen the glaucomatous process, long-term usage of topical glaucoma medications containing BAK should be conducted with caution.
