Sun Yuanyuan, Yu Hongxia, Zhang Jingfei, Yin Ying, Shi Huahong, Wang Xiaorong
State Key Laboratory of Pollution Control and Resources Reuse, School of Environment, Nanjing University, Nanjing, Jiangsu 210093, People's Republic of China.
Chemosphere. 2006 May;63(8):1319-27. doi: 10.1016/j.chemosphere.2005.09.032. Epub 2005 Nov 15.
In this study, laboratory experiment was carried out to determine phenanthrene bioaccumulation, depuration in whole fish and oxidative stress in the liver of freshwater fish Carassius auratus. Fish were exposed to 0.05 mg/l phenanthrene for different periods, while one control group was designated for each exposure group. Some fish after 7 days of exposure were transferred to diluted water. The concentrations of phenanthrene in fish were analyzed by HPLC. Twenty-four hours after the exposure, reactive oxygen species (ROS) were trapped by phenyl-tert-butylnitrone and detected by electron paramagnetic resonance (EPR). The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione-s-transferase (GST) were also determined. The concentrations of phenanthrene in fish increased rapidly shortly after the start of the exposure, reached a maximum level at the 2 days, and then it declined quickly to low-level-steady state. The elimination process of phenanthrene could be divided into two periods-a fast elimination period following a slower loss period. The elimination curve could be fitted mathematically as the sum of two exponential functions according to two-compartment model: C(t)=2.72e(-1.065t) + 0.68e(-0.0364t). The PBN-radical adducts were detected in fish liver samples following the exposure 24h. The hyperfine splitting constants for the PBN-radical adducts were aN = 13.5 G, aH = 1.77 G and g value was 2.0058, which were consistent with those of PBN/()OH. The results indicated that the hydroxyl radical was probably significantly induced during the exposure of phenanthrene, as compared to the control group. The changes of activities of the antioxidant enzymes also were observed. In addition, after fish were removed from phenanthrene exposure, the recovery status of these antioxidant indices was explored. These results clearly indicated phenanthrene could be accumulated in fish and similar redox cyclings were produced, resulting in the changes of the activities of the antioxidant enzymes and the production of ROS with the oxidative stress.
在本研究中，进行了实验室实验以测定菲在淡水鱼鲫鱼体内的生物累积、净化情况以及肝脏中的氧化应激。将鱼暴露于0.05mg/l的菲中不同时间段，每个暴露组设置一个对照组。部分暴露7天后的鱼被转移至稀释水中。通过高效液相色谱法分析鱼体内菲的浓度。暴露24小时后，用苯基叔丁基硝酮捕获活性氧（ROS）并通过电子顺磁共振（EPR）进行检测。还测定了超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽 - S - 转移酶（GST）的活性。鱼体内菲的浓度在暴露开始后不久迅速增加，在第2天达到最高水平，然后迅速下降至低水平稳态。菲的消除过程可分为两个阶段——快速消除阶段和随后的缓慢损失阶段。根据二室模型，消除曲线可以用两个指数函数之和进行数学拟合：C(t)=2.72e(-1.065t)+0.68e(-0.0364t)。暴露24小时后在鱼肝样本中检测到PBN - 自由基加合物。PBN - 自由基加合物的超精细分裂常数为aN = 13.5G，aH = 1.77G，g值为2.0058，与PBN/()OH的一致。结果表明，与对照组相比，菲暴露期间可能显著诱导了羟基自由基。还观察到抗氧化酶活性的变化。此外，在鱼脱离菲暴露后，探究了这些抗氧化指标的恢复情况。这些结果清楚地表明菲可在鱼体内累积并产生类似的氧化还原循环，导致抗氧化酶活性变化以及ROS产生和氧化应激。
