School of Applied Biosciences, Kyungpook National University, Daegu 702-701, South Korea.
Int J Biol Sci. 2013 Apr 13;9(4):343-9. doi: 10.7150/ijbs.5501. Print 2013.
The objective of this study was to find and validate estrogen-related biomarkers from plasma proteins in Oryzias latipes after exposure to an estrogen disrupting compound, α-endosulfan. The acute toxicity of α-endosulfan on O. latipes after 96 h of exposure was 13.72, 16.18, and 22.18 μg L(-1) for the LC10, LC20, and LC50 values, respectively. To confirm estrogenic disturbance by α-endosulfan, the expression level of vitellogenin in the liver of male fishes was measured at the LC10 value, and it was found to be significantly different from the reference group, confirming the estrogenic effect of endosulfan in this concentration range. Proteinchip® array techniques using a weak cation exchange (CM10) and a strong anion exchange proteinchip (Q10) in conjunction with surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) were used to determine plasma proteins of O. latipes differently expressed in response to endosulfan exposure at LC10 and LC20 concentrations. Analysis of protein profiling of the male fish exposed to α-endosulfan detected 48 significantly different protein peaks and the proteins at m/z 2819, 8462, 8860, and 9462 were significantly different (p<0.05). The protein peaks at m/z 2819, 8860, and 9462 were up-regulated and the peak at m/z 8462 was down-regulated. Therefore, these four differentially expressed proteins could be used as biomarkers to rapidly determine a possible risk of endosulfan on aquatic ecosystems, although these are not necessarily produced as a result of endocrine disruption.
本研究旨在寻找并验证暴露于环境雌激素扰乱化合物α-硫丹后，鱼类（Oryzias latipes）血浆蛋白中的雌激素相关生物标志物。α-硫丹对 O. latipes 的 96 小时急性毒性LC10、LC20 和 LC50 值分别为 13.72、16.18 和 22.18μg/L。为了证实α-硫丹的雌激素干扰作用，在 LC10 值下测量了雄性鱼类肝脏中卵黄蛋白原的表达水平，结果与对照组有显著差异，证实了该浓度范围内硫丹的雌激素效应。采用弱阳离子交换（CM10）和强阴离子交换蛋白芯片（Q10）与表面增强激光解吸/电离飞行时间质谱（SELDI-TOF-MS）联用的 Proteinchip® 阵列技术，确定了 O. latipes 血浆蛋白在暴露于 LC10 和 LC20 浓度的硫丹时的不同表达。分析暴露于α-硫丹的雄性鱼类的蛋白质图谱，检测到 48 个差异显著的蛋白质峰，其中 m/z 2819、8462、8860 和 9462 的蛋白质峰差异显著（p<0.05）。m/z 2819、8860 和 9462 的蛋白质峰上调，而 m/z 8462 的蛋白质峰下调。因此，这四个差异表达的蛋白质可以作为生物标志物，快速确定硫丹对水生生态系统可能产生的风险，尽管这些蛋白质不一定是由于内分泌干扰而产生的。
