Luo Yi, Su Yan, Lin Ren-Zhang, Shi Hua-Hong, Wang Xiao-Rong
State Key Laboratory of Pollution Control and Resources Reuse, School of Environment, Nanjing University, Nanjing 210093, China.
Chemosphere. 2006 Nov;65(6):1064-73. doi: 10.1016/j.chemosphere.2006.02.054. Epub 2006 Apr 18.
In the present study, a secondary spin trapping technique was used followed by electron paramagnetic resonance (EPR) analysis, to study the potential of reactive oxygen species (ROS) production after fish (Carassius auratus) were injected i.p. with different doses (50, 100, 200, 250, 500mgkg(-1)) of 2-chlorophenol (2-CP). The ROS signal intensity of the EPR spectrum showed a significant increase (p<0.05, compared with the control) when the 2-CP dose was as low as 50mgkg(-1). There is a good relationship between the 2-CP administered doses and ROS generation. Based on the hyperfine splitting constants and shape of the EPR spectrum, the ROS which was generated in fish liver after intraperitoneal (i.p.) injection of 2-CP was identified as ()OH. SOD and CAT activities were found to be induced at lower doses of 2-CP. GSH levels fell below the control level following all treatments with 2-CP, and GSSG levels changed along with those of GSH. These observations indicated that the fish experienced oxidative stress. The strong positive correlation (r=0.966, p<0.005) between ()OH radical and lipid peroxidation suggested that lipid peroxidation was possibly induced by ()OH. The phase II detoxification enzyme glutathione-S-transferase (GST) may play an important role in 2-CP metabolism or excretion and, consequently, reduce ROS production. This study provides strong evidence that level of ROS is significantly increased in 2-CP stressed fish, and ROS may serve as a potential biomarker to indicate 2-CP contamination.
在本研究中，采用二次自旋捕获技术并结合电子顺磁共振（EPR）分析，以研究腹腔注射不同剂量（50、100、200、250、500mgkg(-1)）2-氯酚（2-CP）后鲫鱼（Carassius auratus）产生活性氧（ROS）的可能性。当2-CP剂量低至50mgkg(-1)时，EPR谱的ROS信号强度显著增加（与对照组相比，p<0.05）。2-CP给药剂量与ROS生成之间存在良好的关系。根据EPR谱的超精细分裂常数和形状，腹腔注射2-CP后在鱼肝中产生的ROS被鉴定为()OH。发现较低剂量的2-CP可诱导超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性。用2-CP进行所有处理后，谷胱甘肽（GSH）水平降至对照水平以下，氧化型谷胱甘肽（GSSG）水平随GSH水平变化。这些观察结果表明鱼经历了氧化应激。()OH自由基与脂质过氧化之间的强正相关（r=0.966，p<\0.005）表明脂质过氧化可能是由()OH诱导的。II相解毒酶谷胱甘肽-S-转移酶（GST）可能在2-CP的代谢或排泄中起重要作用，从而减少ROS的产生。本研究提供了有力证据，表明在2-CP胁迫的鱼中ROS水平显著升高，并且ROS可能作为潜在的生物标志物来指示2-CP污染。
