Schirmer K, Chan A G, Greenberg B M, Dixon D G, Bols N C
Department of Biology, University of Waterloo, Waterloo, ON, N2L 3G1, Canada.
Toxicol In Vitro. 1997 Feb-Apr;11(1-2):107-19. doi: 10.1016/s0887-2333(97)00002-7.
Methodology was developed for quantifying the photocytotoxicity of fluoranthene to a gill cell line from rainbow trout for future use in screening polycyclic aromatic hydrocarbons for their relative photocytotoxicity to fish. Solubilization in a modified culture medium was achieved with and without foetal bovine serum (FBS) and with and without dimethyl sulfoxide (DMSO). FBS caused most of the fluoranthene to remain in solution and blocked photocytotoxicity if present during UV irradiation. DMSO had little effect on fluoranthene distribution in cell cultures but caused cells to be slightly more sensitive to the phototoxicity of fluoranthene. The indicator dyes alamar Blue() and 5-carboxyfluorescein diacetate acetoxymethyl ester were used to quantify cytotoxicity in two different ways-singly in two separate assays, and mixed together in a novel single assay, which saved time and material. With UV irradiation for 2 hr at a photon fluence rate of either 1.4 mumol UV-B/m(2)/sec (UV-A:UV-B, 1.5) or 1.1 mumol UV-B/m(2)/sec (UV-A:UV-B, 9.7), both dyes indicated increasing loss of viability with increasing doses of fluoranthene. EC(50) values ranged from 18 to 44 ng/ml (89-217 nM), with the alamar Blue assay being slightly more sensitive.
已开发出一种方法，用于量化荧蒽对虹鳟鱼鳃细胞系的光细胞毒性，以便未来用于筛选多环芳烃对鱼类的相对光细胞毒性。在添加和不添加胎牛血清（FBS）以及添加和不添加二甲基亚砜（DMSO）的情况下，实现了荧蒽在改良培养基中的溶解。FBS导致大部分荧蒽保留在溶液中，并且如果在紫外线照射期间存在，则会阻断光细胞毒性。DMSO对荧蒽在细胞培养物中的分布影响很小，但会使细胞对荧蒽的光毒性稍微更敏感。指示染料alamar Blue（）和5-羧基荧光素二乙酸酯乙酰氧基甲酯用于以两种不同方式量化细胞毒性——在两个单独的试验中单独使用，以及在一种新颖的单一试验中混合使用，这节省了时间和材料。在光子通量率为1.4 μmol UV-B/m²/秒（UV-A:UV-B，1.5）或1.1 μmol UV-B/m²/秒（UV-A:UV-B，9.7）的条件下进行2小时紫外线照射，两种染料均表明随着荧蒽剂量增加，细胞活力损失增加。半数有效浓度（EC₅₀）值范围为18至44 ng/ml（89至217 nM），alamar Blue试验稍微更敏感。
