Godard C A J, Wise S S, Kelly R S, Goodale B, Kraus S, Romano T, O'Hara T, Wise J P
Wise Laboratory of Environmental and Genetic Toxicology, University of Southern Maine, Portland, ME 04104, USA.
Mar Environ Res. 2006 Jul;62 Suppl:S20-4. doi: 10.1016/j.marenvres.2006.04.016. Epub 2006 Apr 15.
The North Atlantic right whale (NARW) is one of the most endangered great whales. The NARW population consists of only about 300 individuals and is reproducing at an insufficient rate. There is growing concern about the potential effects of environmental contaminants on the reproductive and overall health of NARW. High contaminant burdens can accumulate in tissues of great whales but toxicological studies of their effects are limited due to legal, logistical and ethical restrictions and specific in vitro models are critically needed. Cell lines from NARW skin and internal organs were previously created in our laboratory. In this study, skin, testis and lung primary fibroblast cell lines were exposed to benzo[a]pyrene (BP) as part of a multi-chemical toxicity testing project in NARW. Cells were exposed for 24-72 h to 10 nM-10 microM BP dissolved in dimethylsulfoxide. Cytotoxicity was measured with a clonogenic assay using standard methods. Some cytotoxicity was observed after 24 h, the highest concentration (10 microM BP) resulting in 77, 74 and 51 percent relative survival in testis, skin and lung cells, respectively, and indicating a higher cytotoxicity in the lung (p < 0.05). After 48 and 72-h exposure, 10 microM BP resulted in 24 and 3, 74 and 27, and 42 and 23 percent relative survival in testis, skin and lung cells, respectively. Cytotoxicity significantly increased with exposure time in all three tissues (p < 0.05 for skin and p < 0.01 for lung and testis), suggesting metabolic activation of BP in the three organs. Fibroblast cytotoxicity observed in the testis was higher than that observed either in the skin or lung after 48 h (p < 0.01) and was close to 100% after 72 h, warranting further investigation of the potential effects of PAHs on reproductive health.
北大西洋露脊鲸是最濒危的大型鲸鱼之一。北大西洋露脊鲸种群仅约由300头个体组成,繁殖率不足。人们越来越关注环境污染物对北大西洋露脊鲸繁殖和整体健康的潜在影响。高污染物负荷会在大型鲸鱼的组织中积累,但由于法律、后勤和伦理限制,关于其影响的毒理学研究有限,因此迫切需要特定的体外模型。我们实验室之前已创建了来自北大西洋露脊鲸皮肤和内脏器官的细胞系。在本研究中,作为北大西洋露脊鲸多化学物质毒性测试项目的一部分,皮肤、睾丸和肺原代成纤维细胞系被暴露于苯并[a]芘(BP)。细胞暴露于溶解在二甲基亚砜中的10 nM - 10 microM BP 24至72小时。使用标准方法通过克隆形成试验测量细胞毒性。24小时后观察到一些细胞毒性,最高浓度(10 microM BP)分别导致睾丸、皮肤和肺细胞的相对存活率为77%、74%和51%,表明肺中的细胞毒性更高(p < 0.05)。暴露48小时和72小时后,10 microM BP分别导致睾丸、皮肤和肺细胞的相对存活率为24%和3%、74%和27%、42%和23%。所有三个组织中的细胞毒性均随暴露时间显著增加(皮肤p < 0.05,肺和睾丸p < 0.01),表明BP在这三个器官中发生了代谢活化。48小时后在睾丸中观察到的成纤维细胞毒性高于在皮肤或肺中观察到的毒性(p < 0.01),72小时后接近100%,这值得进一步研究多环芳烃对生殖健康的潜在影响。