Department of Ophthalmology, School of Medicine, University of California, Irvine, CA 92868, USA.
Invest Ophthalmol Vis Sci. 2010 May;51(5):2601-7. doi: 10.1167/iovs.09-4121. Epub 2009 Dec 3.
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-microM 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 microM), 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.
研究几种药物（抑制剂）对体外人视网膜色素上皮（ARPE-19）细胞中苯并（e）芘（B（e）P）诱导的细胞死亡和凋亡的抑制作用。
在暴露于 B（e）P 之前，用不同浓度的不同类抑制剂（钙蛋白酶、苄基异硫氰酸酯[BITC]、辛伐他汀、表儿茶素、染料木黄酮、白藜芦醇和盐酸美金刚）预处理 ARPE-19 细胞。通过台盼蓝染料排除试验测定细胞活力（CV）。通过荧光染料测定法测定 caspase-3/7 和 caspase-9 的活性。用 2'，7'-二氯二氢荧光素二乙酸酯染料测定法测定活性氧/氮物种（ROS/RNS）的产生。
在 30μM 浓度下，染料木黄酮、白藜芦醇和盐酸美金刚抑制剂能够显著逆转 ARPE-19 细胞培养物中细胞活力的丧失、caspase-3/7 和 caspase-9 的激活以及 ROS/RNS 的产生。盐酸美金刚是最有效的抑制剂，而染料木黄酮是阻断 B（e）P 诱导作用的最无效抑制剂。钙蛋白酶、BITC、辛伐他汀和表儿茶素不能逆转 B（e）P 处理的 ARPE-19 细胞中的细胞活力丧失。事实上，在所研究的浓度（15、30、45μM）下，与单独用 B（e）P 处理的培养物相比，BITC 加 B（e）P 处理的培养物显示出显著更低的细胞活力，表明 BITC 相关的毒性。
染料木黄酮、白藜芦醇和盐酸美金刚可以逆转 B（e）P 产生的凋亡和氧化剂产生，B（e）P 是吸烟的有毒元素。这些抑制剂可能对与 RPE 细胞丧失相关的视网膜疾病有益。
