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Saffar Mansoor, Navin Gupta, A. Jayaprakash Patil, Maria Fernanda Estrago-Franco, Claudio Ramirez, Rafael Migon, Ashish Sapkal, Baruch D. Kuppermann, M. Cristina Kenney; Inhibition of Apoptosis in Human Retinal Pigment Epithelial Cells Treated with Benzo(e)Pyrene, a Toxic Component of Cigarette Smoke. Invest. Ophthalmol. Vis. Sci. 2010;51(5):2601-2607. doi: https://doi.org/10.1167/iovs.09-4121.
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To study the inhibitory effects of some agents or drugs (inhibitors) on benzo(e)pyrene (B(e)P)-induced cell death and apoptosis on human retinal pigment epithelial (ARPE-19) cells in vitro.
ARPE-19 cells were pretreated with varying concentrations of different classes of inhibitors (calpain, benzyl isothiocyanate [BITC], simvastatin, epicatechin, genistein, resveratrol, and memantine) before B(e)P exposure. Cell viability (CV) was determined by a trypan blue dye-exclusion assay. Caspase-3/7 and caspase-9 activities were measured by fluorochrome assays. The production of reactive oxygen/nitrogen species (ROS/RNS) was measured with 2′,7′-dicholorodihydrofluorescein diacetate dye assay.
At 30-μM concentrations, the genistein, resveratrol, and memantine inhibitors were able to reverse significantly the loss of cell viability, the activation of caspase-3/7 and caspase-9, and the production of ROS/RNS in ARPE-19 cell cultures. Memantine was the most potent and genistein was the least effective inhibitor in blocking the B(e)P-induced effects. Calpain, BITC, simvastatin, and epicatechin did not reverse the loss of cell viability in B(e)P-treated ARPE-19 cells. As a matter of fact, at the concentrations studied (15, 30, 45 μM), the BITC plus B(e)P-treated cultures showed significantly lower cell viability than the B(e)P-treated culture alone, suggesting BITC-related toxicity.
Genistein, resveratrol, and memantine can reverse the apoptosis and oxidant production generated by B(e)P, a toxic element of smoking. These inhibitors may be beneficial against retinal diseases associated with the loss of RPE cells.
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