Ahmad Iqbal, Maria V L, Oliveira M, Pacheco M, Santos M A
Animal Physiology/Ecotoxicology Sector, Department of Biology, University of Aveiro, 3810-193 Aveiro, Portugal.
Mutat Res. 2006 Sep 19;608(1):16-28. doi: 10.1016/j.mrgentox.2006.04.020. Epub 2006 Jun 19.
Fish in the aquatic environment can be subjected to a multipollution state and the occurrence of sequential exposures is an important aspect of eco-toxicological research. In this context, a preceding exposure can affect a toxic response to a subsequent exposure. Therefore, the current study was based on sequential exposures, viz. to a PAH-like compound (beta-naphthoflavone, BNF) followed by a heavy metal (chromium, Cr), focusing on the assessment of oxidative stress responses and their role in induction of genotoxicity. Oxidative stress responses in gill and kidney were investigated in European eel (Anguilla anguilla L.), and measured as lipid peroxidation (LPO), glutathione peroxidase (GPX), catalase (CAT) and glutathione S-transferase (GST) activity, and reduced glutathione (GSH) concentration, whereas genotoxicity was measured as DNA strand breakage. Fish were exposed for 24 h to two Cr concentrations (100 microM, 1 mM), with or without pre-exposure to BNF (2.7 microM, 24 h). In gill, a GSH decrease was observed along with loss of DNA integrity at all exposure conditions except at the lowest Cr concentration, showing a crucial role of GSH over genotoxicity. Moreover, sporadic induction of antioxidant enzymes was not effective in the protection against genotoxicity. However, a different mechanism seems to occur in kidney, since the loss of DNA integrity detected for all exposed groups was not accompanied by alterations in antioxidant levels. With regards to peroxidative damage, both organs showed an LPO increase after sequential exposure to BNF and 100 microM Cr. However, no association between LPO induction and antioxidant responses could be established, showing that LPO is not predictable solely on the basis of antioxidant depletion. The interference of BNF pre-exposure with the response of organs to Cr showed a marked dependence on the Cr concentration. Gill showed synergistic effects on LPO and GPX increase, as well as on CAT and GSH decrease for the lowest Cr concentration. However, for the highest concentration an additive effect on decrease of DNA integrity and an antagonistic effect on the increase of GPX were observed. In kidney, synergistic effects were evident on LPO increase and GSH decrease for the lowest Cr concentration, as well as on CAT and GST decrease for the highest concentration. In contrast, an antagonistic action was observed on DNA integrity loss for both Cr concentrations. The current results are relevant in assessing the interactions of PAHs and metals and contribute to a better knowledge about oxidative stress and mechanisms of genotoxicity in fish.
水生环境中的鱼类可能会处于多重污染状态,连续暴露的情况是生态毒理学研究的一个重要方面。在这种背景下,先前的暴露可能会影响对后续暴露的毒性反应。因此,本研究基于连续暴露,即先暴露于一种多环芳烃类化合物(β-萘黄酮,BNF),随后暴露于一种重金属(铬,Cr),重点评估氧化应激反应及其在遗传毒性诱导中的作用。在欧洲鳗鲡(Anguilla anguilla L.)中研究了鳃和肾脏中的氧化应激反应,并以脂质过氧化(LPO)、谷胱甘肽过氧化物酶(GPX)、过氧化氢酶(CAT)和谷胱甘肽S-转移酶(GST)活性以及还原型谷胱甘肽(GSH)浓度来衡量,而遗传毒性则以DNA链断裂来衡量。将鱼暴露于两种Cr浓度(100 microM、1 mM)下24小时,有或没有预先暴露于BNF(2.7 microM,24小时)。在鳃中,除了最低Cr浓度外,在所有暴露条件下均观察到GSH下降以及DNA完整性丧失,表明GSH对遗传毒性起着关键作用。此外,抗氧化酶的零星诱导在防止遗传毒性方面并不有效。然而,肾脏中似乎发生了不同的机制,因为在所有暴露组中检测到的DNA完整性丧失并未伴随着抗氧化水平的改变。关于过氧化损伤,在连续暴露于BNF和100 microM Cr后,两个器官的LPO均增加。然而,无法确定LPO诱导与抗氧化反应之间的关联,表明LPO不能仅基于抗氧化剂消耗来预测。预先暴露的BNF对器官对Cr反应的干扰显示出对Cr浓度的明显依赖性。对于最低Cr浓度,鳃对LPO和GPX增加以及CAT和GSH下降表现出协同作用。然而,对于最高浓度,观察到对DNA完整性下降有相加作用,对GPX增加有拮抗作用。在肾脏中,对于最低Cr浓度,对LPO增加和GSH下降有明显的协同作用,对于最高浓度,对CAT和GST下降有协同作用。相反,对于两种Cr浓度,在DNA完整性丧失方面均观察到拮抗作用。当前结果对于评估多环芳烃和金属的相互作用具有重要意义,并有助于更好地了解鱼类中的氧化应激和遗传毒性机制。