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T. H. Huynh, A. Sharma, A. L. Gramajo, A. Jayaprakash Patil, M. F. Estrago Franco, M. Chwa, G. M. Seigel, B. D. Kuppermann, M. C. Kenney; Effect of Hydroquinone, a Toxicant in Cigarette Smoke, on Human Retinal Pigment Epithelial Cells, Rat Retinal Neurosensory Cells and Human Microvascular Endothelial Cells in vitro. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5936.
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© ARVO (1962-2015); The Authors (2016-present)
To determine the epidemiologic association between cigarette smoke and age-related macular degeneration, we examined the effects of Hydroquinone (HQ), a toxic element present in high concentration in cigarette-smoke related tar, on human retinal pigment epithelial cells (ARPE-19), rat neurosensory cells (R28) and human microvascular endothelial (HMVEC) cells.
ARPE-19, R28, and HMVEC cells were grown in tissue culture and treated with four different concentrations of HQ (50µM, 100µM, 200µM, 500µM). Trypan blue cell viability assay, caspase-3/7 assay, and DNA laddering were performed to analyze the molecular effects of HQ on these cells.
The mean cell viability percentages of ARPE-19 cells after 24 hours exposure to HQ 50µM, 100µM, 200µM, 500µM were 95.70±0.42 (P>0.05), 52.95±2.051 (P<0.001), 33.50± 3.53 (P<0.001) and 26.60±1.56 (P<0.001) respectively compared to 500µM equivalent DMSO controls 96.250±0.78.The mean cell viability percentages of R28 cells after 24 hours exposure to HQ 50µM, 100µM, 200µM, 500µM were 67.3± 2.9 (P<0.01), 35.70±3.252 (P<0.001), 26.20± 1.13(P <0.001) and 14.15±1.20 (P<0.001) respectively compared to 500µM equivalent DMSO controls 78.56 ±2.90.The mean cell viability percentages of HMVEC after 24 hours exposure to HQ 50µM, 100µM, 200µM, 500µM were 93.80±0.92 (P>0.05), 37.52±1.93 (P<0.001), 25.93± 2.17(P <0.001) and 11.73±2.27 (P<0.001) respectively compared to 500µM equivalent DMSO controls 91.50± 2.19.At 50µM HQ, the R28 cells showed a significant decrease of cell viability 67.3± 2.9 (P<0.01), while ARPE-19 and HMVEC cultures were similar to the DMSO treated controls. All cells showed loss of cell viability at 100µM, 200µM and 500µM HQ. There was no significant increase in caspase-3/7 activities in all three cell lines at all concentrations tested. DNA laddering did not show any banding pattern.
This study suggests that R28 cells are more sensitive to HQ than human ARPE-19 and HMVEC cells. The mechanism of cell death was necrosis in all three cells lines.
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