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北极海鸟体内多氯联苯的生物转化:在转录、翻译和活性水平上对 I 期和 II 期途径的特征描述。

Biotransformation of PCBs in Arctic seabirds: characterization of phase I and II pathways at transcriptional, translational and activity levels.

机构信息

Norwegian Polar Institute, 9296 Tromsø, Norway.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2010 Jun;152(1):34-41. doi: 10.1016/j.cbpc.2010.02.009. Epub 2010 Feb 20.

Abstract

Arctic seabirds are exposed to a wide range of halogenated organic contaminants (HOCs). Exposure occurs mainly through food intake, and many pollutants accumulate in lipid-rich tissues. Little is known about how HOCs are biotransformed in arctic seabirds. In this study, we characterized biotransformation enzymes in chicks of northern fulmars (Fulmarus glacialis) and black-legged kittiwakes (Rissa tridactyla) from Kongsfjorden (Svalbard, Norway). Phase I and II enzymes were analyzed at the transcriptional, translational and activity levels. For gene expression patterns, quantitative polymerase chain reactions (qPCR), using gene-sequence primers, were performed. Protein levels were analyzed using immunochemical assays of western blot with commercially available antibodies. Liver samples were analyzed for phase I and II enzyme activities using a variety of substrates including ethoxyresorufin (cytochrome (CYP)1A1/1A2), pentoxyresorufin (CYP2B), methoxyresorufin (CYP1A), benzyloxyresorufin (CYP3A), testosterone (CYP3A/CYP2B), 1-chloro-2,4-nitrobenzene (CDNB) (glutathione S-transferase (GST)) and 4-nitrophenol (uridine diphosphate glucuronyltransferase (UDPGT)). In addition, the hydroxylated (OH-) polychlorinated biphenyls (PCBs) were analyzed in the blood, liver and brain tissue, whereas the methylsulfone (MeSO(2)-) PCBs were analyzed in liver tissue. Results indicated the presence of phase I (CYP1A4/CYP1A5, CYP2B, and CYP3A) and phase II (GST and UDPGT) enzymes at the activity, protein and/or mRNA level in both species. Northern fulmar chicks had higher enzyme activity than black-legged kittiwake chicks. This in combination with the higher SigmaOH-PCB to parent PCB ratios suggests that northern fulmar chicks have a different biotransformation capacity than black-legged kittiwake chicks.

摘要

北极海鸟暴露于广泛的卤代有机污染物(HOCs)中。暴露主要通过食物摄入发生,并且许多污染物在富含脂质的组织中积累。关于 HOCs 在北极海鸟中如何被生物转化知之甚少。在这项研究中,我们描述了来自 Kongsfjorden(挪威斯瓦尔巴特群岛)的北方管鼻藿(Fulmarus glacialis)和黑腿海鸥(Rissa tridactyla)雏鸟中的生物转化酶。在转录、翻译和活性水平分析了 I 相和 II 相酶。对于基因表达模式,使用基因序列引物进行了定量聚合酶链反应(qPCR)。使用商业上可用的抗体进行 Western blot 的免疫化学测定来分析蛋白质水平。使用各种底物分析肝样品中的 I 相和 II 相酶活性,包括乙氧基resorufin(细胞色素(CYP)1A1/1A2)、戊氧基resorufin(CYP2B)、甲氧基resorufin(CYP1A)、苄氧基resorufin(CYP3A)、睾酮(CYP3A/CYP2B)、1-氯-2,4-二硝基苯(CDNB)(谷胱甘肽 S-转移酶(GST))和 4-硝基苯酚(尿苷二磷酸葡萄糖醛酸转移酶(UDPGT))。此外,在血液、肝脏和脑组织中分析了羟基化(OH-)多氯联苯(PCBs),而在肝脏组织中分析了甲基砜(MeSO 2-)PCBs。结果表明,在这两个物种中,在活性、蛋白质和/或 mRNA 水平上均存在 I 相(CYP1A4/CYP1A5、CYP2B 和 CYP3A)和 II 相(GST 和 UDPGT)酶。北方管鼻藿雏鸟的酶活性高于黑腿海鸥雏鸟。这与较高的 SigmaOH-PCB 与母体 PCB 比值相结合表明,北方管鼻藿雏鸟具有与黑腿海鸥雏鸟不同的生物转化能力。

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