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毒死蜱对金鱼(Carassius auratus)CYP3A cDNA转录、乙酰胆碱酯酶活性及氧化应激反应的影响。

Effects of chlorpyrifos on the transcription of CYP3A cDNA, activity of acetylcholinesterase, and oxidative stress response of goldfish (Carassius auratus).

作者信息

Ma Junguo, Liu Yang, Niu Daichun, Li Xiaoyu

机构信息

College of Life Science, Henan Normal University, Xinxiang, Henan, 453007, China.

出版信息

Environ Toxicol. 2015 Apr;30(4):422-9. doi: 10.1002/tox.21918. Epub 2013 Nov 4.

Abstract

Chlorpyrifos (CPF) is the widely used organophosphate pesticide in agriculture throughout the world. It has been found that CPF is relatively safe to human but highly toxic to fish. In this study, acute toxicity of CPF on goldfish was determined and then the transcription of goldfish cytochrome P450 (CYP) 3A was evaluated after 96 h of CPF exposure at concentrations of 15.3 [1/10 50% lethal concentration (LC50 )] or 51 μg L(-1) (1/3 LC50 ) of CPF. Meanwhile, the enzymatic activities of acetylcholinesterase (AChE), superoxide dismutase (SOD), and catalase (CAT), total antioxidant activity (T-AOC), and the contents of malondialdehyde (MDA) in the liver or brain of goldfish were also determined. The results of acute toxicity testing showed that the 96-h LC50 of CPF to the goldfish was 153 μg L(-1) . Moreover, a length sequence of 1243 bp CYP3A cDNA encoding for 413 amino acids from goldfish liver was cloned. Polymerase chain reaction results reveal that CPF exposure downregulates CYP 3A transcription in goldfish liver, suggesting that goldfish CYP 3A may be not involved in CPF bioactivation. Finally, the results of biochemical assays indicate that 96 h of CPF exposure remarkably inhibits AChE activity in fish liver or brain, alters hepatic antioxidant enzyme activities, decreases brain T-AOC, and causes lipid peroxidation in fish liver. These results suggest that oxidative stress might be involved in CPF toxicity on goldfish.

摘要

毒死蜱(CPF)是全球农业中广泛使用的有机磷农药。已发现CPF对人类相对安全,但对鱼类剧毒。在本研究中,测定了CPF对金鱼的急性毒性,然后在浓度为15.3 [1/10半数致死浓度(LC50)]或51 μg L(-1)(1/3 LC50)的CPF暴露96小时后,评估了金鱼细胞色素P450(CYP)3A的转录情况。同时,还测定了金鱼肝脏或大脑中乙酰胆碱酯酶(AChE)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)的酶活性、总抗氧化活性(T-AOC)以及丙二醛(MDA)的含量。急性毒性测试结果表明,CPF对金鱼的96小时LC50为153 μg L(-1)。此外,克隆了金鱼肝脏中编码413个氨基酸的1243 bp CYP3A cDNA的长度序列。聚合酶链反应结果显示,CPF暴露下调了金鱼肝脏中CYP 3A的转录,表明金鱼CYP 3A可能不参与CPF的生物活化。最后,生化分析结果表明,CPF暴露96小时显著抑制了鱼肝或脑中的AChE活性,改变了肝脏抗氧化酶活性,降低了脑T-AOC,并导致鱼肝脂质过氧化。这些结果表明,氧化应激可能参与了CPF对金鱼的毒性作用。

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