Gong Yinhan, Chin Hong Soon, Lim Lis Sa Elissa, Loy Chong Jin, Obbard Jeffrey P, Yong E L
Tropical Marine Science Institute, National University of Singapore, Republic of Singapore.
Environ Health Perspect. 2003 Sep;111(12):1448-53. doi: 10.1289/ehp.6139.
Abnormal sexual differentiation and other reproductive abnormalities in marine animals indicate the presence in seawater of endocrine-disrupting compounds (EDCs) that perturb the function of the sex hormone signaling pathways. However, most studies to date have reported on EDC effects in freshwater and sewage samples, and there is a paucity of bioassay data on the effects of EDCs in marine waters. Our aims in this study were to devise robust methodologies suitable for extracting potential EDCs and to measure their summated effects on activities of androgen receptors (ARs) and estrogen receptors (ER-alpha and ER-beta) in marine samples from Singapore's coastal waters. In this study, we examined the ability of C18, hydrophilic and lipophilic balance, and diol cartridges to extract potential EDCs from seawater samples. Extracts from C18 cartridges exhibited the highest sex hormone bioactivities in reporter gene assays based on a human cell line expressing AR, ER-alpha, and ER-beta. Examination of extracts from 20 coastal locations showed high androgenic and estrogenic agonist activities in confined clusters closest to the main island of Singapore. Sex hormone activity declined rapidly in clusters farther from the main coastline and in more open waters. Unexpectedly, surface and mid-depth samples from the confined high-activity clusters, in the presence of hormone, exhibited AR and ER-alpha activities that were 200-900% higher than those observed for the cognate hormone alone. This enhanced sex hormone activity suggests that analyses of complex seawater mixtures may uncover unusual bioactivities that may not be obvious by studying individual compounds. Our data present a "snapshot" of the sex hormone disruptor activity in Singapore's marine environment and indicate that C18 extraction for EDCs used in conjunction with reporter gene bioassays represents a robust and sensitive methodology for measuring summated androgenic and estrogenic activities in seawater.
海洋动物的异常性别分化及其他生殖异常表明海水中存在扰乱性激素信号通路功能的内分泌干扰化合物（EDCs）。然而，迄今为止，大多数研究报告的是EDCs在淡水和污水样本中的影响，关于EDCs对海水影响的生物测定数据较少。我们在本研究中的目标是设计出适用于提取潜在EDCs的可靠方法，并测量它们对来自新加坡沿海水域海洋样本中雄激素受体（ARs）和雌激素受体（ER-α和ER-β）活性的综合影响。在本研究中，我们考察了C18、亲水亲油平衡柱和二醇柱从海水样本中提取潜在EDCs的能力。在基于表达AR、ER-α和ER-β的人类细胞系的报告基因测定中，C18柱提取物表现出最高的性激素生物活性。对20个沿海地点的提取物进行检测发现，在最靠近新加坡主岛的有限区域内存在较高的雄激素和雌激素激动剂活性。在离主要海岸线较远的区域和更开阔的水域中，性激素活性迅速下降。出乎意料的是，在存在激素的情况下，来自有限高活性区域的表层和中层深度样本所表现出的AR和ER-α活性比单独观察到的同源激素高出200 - 900%。这种增强的性激素活性表明，对复杂海水混合物的分析可能会揭示出一些通过研究单个化合物可能不明显的异常生物活性。我们的数据呈现了新加坡海洋环境中性激素干扰物活性的“快照”，并表明结合报告基因生物测定使用的C18柱提取EDCs是一种测量海水中雄激素和雌激素综合活性的可靠且灵敏的方法。
