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목적 : The goal of this study was to determine what effect chronic exposure of low dose Benzalkonium Chloride (BAK) can induce on trabecular meshwork cells and to investigate the role of angiogenin (ANG) on cellular senescence.
방법 : Trabecular meshwork cells were treated with 0.0005%, 0.00075%, 0.001%, 0.0025% BAK for 10 minutes, then transferred to a new medium for 24 hours. For ANG and BAK combined treatment, 5ug/mL ANG was applied for 24 hours in advance of BAK in the above experimental design. This process was repeated 3 times. Cell survival was assessed through MTT assay and SA-β-gal staining was performed to quantify cellular senescence in comparison to a control group. To understand the molecular pathway involved, Western blot was performed to see whether the expressions of cell cycle regulators were affected.
결과 : Two concentrations (0.0005% and 0.00075%) showed persistent cell viability and were chosen for further experiments accordingly. In SA-β-gal staining, when compared to control, cells that were treated with BAK showed considerable increase in senescent expression but cells that were treated with BAK and ANG combined did not. Western blot analysis with BAK alone resulted in decreased expressions of cyclin E/CDK2, and increased expressions of the upper stream control molecules, p53 and p21. Western blot for BAK and ANG treated cells resulted in decreased expression of p53.
결론 : Chronic exposure to low dose BAK, a similar environment to the anterior chamber in glaucoma, seems to accelerate cell senescence through cell cycle arrest. Angiogenin, on the other hand, may have protective effect against BAK-induced cellular senescence in trabecular meshwork cells.
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