Contamination with retinoic acid receptor agonists in two rivers in the Kinki region of Japan.
Contamination with retinoic acid receptor agonists in two rivers in the Kinki region of Japan.
复制标题
DOI：
10.1016/j.watres.2010.01.005
复制
发表时间：
2010-04
期刊：
影响因子：
12.8
通讯作者：
D. Inoue;Koki Nakama;K. Sawada;Taro Watanabe;Mai Takagi;K. Sei;Min Yang;J. Hirotsuji;Jianying Hu;J. Nishikawa;T. Nakanishi;M. Ike
中科院分区：
文献类型：
--
作者：
D. Inoue;Koki Nakama;K. Sawada;Taro Watanabe;Mai Takagi;K. Sei;Min Yang;J. Hirotsuji;Jianying Hu;J. Nishikawa;T. Nakanishi;M. Ike
本研究旨在研究日本近畿地区的琵琶湖淀川和伊那川中对人维甲酸受体（RAR）α的激动活性。为了实现这一点，酵母双杂交法被用来阐明空间和时间的变化和潜在来源的RARα激动剂污染的河流流域。在两条河流不同时期采集的地表水样品中普遍检测到RARα激动活性，其中在琵琶湖淀川和伊那川中检测到的全反式维甲酸（atRA）当量最大，分别为47. 6 ng-atRA/L和23. 5 ng-atRA/L。结果表明，RARα激动剂在河流中普遍存在且分布广泛。通过对各河流沿着20个站点RARα和雌激素受体α激动剂活性的比较研究，发现RARα激动剂污染的空间变化规律与雌激素类化合物污染的空间变化规律完全不同。这表明，城市污水处理厂的污水是雌激素化合物的主要来源，似乎不是河流中RARα激动剂污染的原因。在生物测定的指导下，使用高效液相色谱法（HPLC）进行分级，从河水样品中发现了两个生物活性级分，表明河流中至少存在两种RARα激动剂。尽管本研究中未完成鉴定主要生物活性组分中RARα激动剂的试验，但HPLC分析中保留时间的比较和液相色谱-质谱分析定量显示，导致RARα激动活性的主要致病污染物不是RA（天然RAR配体）和4-氧代-RA，而4-氧代-RA被鉴定为中国北京的污水中的主要RAR激动剂。这些结果表明，在河流中存在未知的高活性RARα激动剂。
This study was conducted to investigate the agonistic activity against human retinoic acid receptor (RAR) α in the Lake Biwa–Yodo River and the Ina River in the Kinki region of Japan. To accomplish this, a yeast two-hybrid assay was used to elucidate the spatial and temporal variations and potential sources of RARα agonist contamination in the river basins. RARα agonistic activity was commonly detected in the surface water samples collected along two rivers at different periods, with maximum all-trans retinoic acid (atRA) equivalents of 47.6ng-atRA/L and 23.5ng-atRA/L being observed in Lake Biwa–Yodo River and Ina River, respectively. The results indicated that RARα agonists are always present and widespread in the rivers. Comparative investigation of RARα and estrogen receptor α agonistic activities at 20 stations along each river revealed that the spatial variation pattern of RARα agonist contamination was entirely different from that of the estrogenic compound contamination. This suggests that the effluent from municipal wastewater treatment plants, a primary source of estrogenic compounds, seemed not to be the cause of RARα agonist contamination in the rivers. Fractionation using high performance liquid chromatography (HPLC) directed by the bioassay found two bioactive fractions from river water samples, suggesting the presence of at least two RARα agonists in the rivers. Although a trial conducted to identify RARα agonists in the major bioactive fraction was not completed as part of this study, comparison of retention times in HPLC analysis and quantification with liquid chromatography–mass spectrometry analysis revealed that the major causative contaminants responsible for the RARα agonistic activity were not RAs (natural RAR ligands) and 4-oxo-RAs, while 4-oxo-RAs were identified as the major RAR agonists in sewage in Beijing, China. These findings suggest that there are unknown RARα agonists with high activity in the rivers.