Zearalenone activates pregnane X receptor, constitutive androstane receptor and aryl hydrocarbon receptor and corresponding phase I target genes mRNA in primary cultures of human hepatocytes
Zearalenone activates pregnane X receptor, constitutive androstane receptor and aryl hydrocarbon receptor and corresponding phase I target genes mRNA in primary cultures of human hepatocytes
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DOI：
10.1016/j.etap.2010.09.008
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发表时间：
2011-01-01
影响因子：
4.3
通讯作者：
Hassen, Wafa
中科院分区：
文献类型：
--
作者：
Ayed-Boussema, Imen;Pascussi, Jean Marc;Hassen, Wafa
真菌毒素玉米赤霉烯酮（ZEN）是世界范围内发现的谷物和谷物污染物。ZEN的亚慢性和慢性毒性主要表现为不同哺乳动物物种的生殖障碍，这使得ZEN被确定为哺乳动物内分泌干扰物。在过去30年的ZEN生物转化研究中，该毒素被认为仅经历还原代谢，在几种物种中产生玉米赤霉烯醇的α-和β-异构体。然而，最近的研究已经注意到，真菌雌激素易于氧化代谢，导致ZEN的羟基化，通过不同的细胞色素P450（CYP）亚型的参与。本研究的目的是进一步探讨ZEN对人肝细胞原代培养物中某些CYP的调节作用。为此目的，我们首次采用真实的时间RT-PCR监测了ZEN对已知参与某些CYP调节的核受体--雌酮X受体（PXR）、组成型雄烷受体（CAR）和芳香烃受体（AhR）mRNA水平的影响。第二次，我们观察了ZEN对PXR、CAR和AhR相应I期靶基因（CYP 3A 4、CYP 3A 5、CYP 2B 6、CYP 2C 9、CYP 1A 1和CYP 1A 2）表达的影响。最后，我们在用毒素处理并在hPXR载体存在下用p-CYP 3A 4-Luc瞬时转染或用p-CYPA 1-Luc转染的HepG 2中实现了荧光素酶测定。我们的结果清楚地表明，ZEN激活人类PXR，CAR和AhR mRNA水平，除了它们的一些I期靶基因，主要是CYP 3A 4，CYP 2B 6和CYP 1A 1，在较小程度上CYP 3A 5和CYP 2C 9，ZEN浓度低至0.1 μ M。(C)2010 Elsevier B. V.保留所有权利。
The mycotoxin zearalenone (ZEN) is found worldwide as a contaminant in cereals and grains. ZEN subchronic and chronic toxicities are dominated by reproductive disorders in different mammalian species which have made ZEN established mammalian endocrine disrupter. Over the last 30 years of ZEN biotransformation study, the toxin was thought to undergo reductive metabolism only, with the generation in several species of alpha- and beta-isomers of zearalenol. However, recent investigations have noticed that the mycoestrogen is prone to oxidative metabolism leading to hydroxylation of ZEN though the involvement of different cytochromes P450 (CYPs) isoforms. The aim of the present study was to further explore the effect of ZEN on regulation of some CYPs using primary cultures of human hepatocytes. For this aim, using real time RT-PCR, we monitored in a first time, the effect of ZEN on mRNA levels of pregnane X receptor (PXR), constitutive androstane receptor (CAR) and aryl hydrocarbon receptor (AhR), nuclear receptors known to be involved in the regulation of some CYPs. In a second time, we looked for ZEN effect on expression of PXR, CAR and AhR corresponding phase I target genes (CYP3A4, CYP3A5, CYP2B6, CYP2C9, CYP1A1 and CYP1A2). Finally, we realised the luciferase assay in HepG2 treated with the toxin and transiently transfected with p-CYP3A4-Luc in the presence of a hPXR vector or transfected with p-CYPA1-Luc. Our results clearly showed that ZEN activated human PXR, CAR and AhR mRNA levels in addition to some of their phase I target genes mainly CYP3A4, CYP2B6 and CYP1A1 and at lesser extent CYP3A5 and CYP2C9 at ZEN concentrations as low as 0.1 mu M. (C) 2010 Elsevier B.V. All rights reserved.