Structural influence of isothiocyanates on expression of cytochrome P450, phase II enzymes, and activation of Nrf2 in primary rat hepatocytes
Structural influence of isothiocyanates on expression of cytochrome P450, phase II enzymes, and activation of Nrf2 in primary rat hepatocytes
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DOI：
10.1016/j.fct.2012.05.044
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发表时间：
2012-08-01
影响因子：
4.3
通讯作者：
Longo, V.
中科院分区：
文献类型：
--
作者：
La Marca, M.;Beffy, P.;Longo, V.
使用大鼠肝细胞的原代培养物来研究是否以及如何从水解芥子油苷中衍生出八种具有不同化学结构的异硫氰酸酯（ITC）（芳香族苯甲基、4-羟基苯甲基、苯乙基异硫氰酸酯和脂肪族烯丙酯、napin、iberin、raphasatin 异硫氰酸酯和萝卜硫素）。 能够调节细胞色素 P450 (CYP) 和抗氧化/解毒酶并激活 Nrf2 转录因子。处理 24 小时后，40 μM 的芳香族 ITC 显着增加了 CYP1A1 和 1A2 mRNA 的转录，并增加了相关的乙氧基试卤灵 O-脱乙基酶 (EROD) 活性。相比之下，脂肪族ITC (40 μM) 降低了CYP1A1 和1A2 转录以及相应的EROD 活性。相同的处理还引起了 CYP3A2 的惊人且相似的转录抑制，以及响应所有测试的 ITC 的相应的苄氧基喹啉脱苄酶活性。在相同的培养条件下，大多数抗氧化/解毒酶被40 mu M ITCs显着上调。特别是，NAD(P)H:醌氧化还原酶和血红素加氧酶-1均被所有ITC在转录、蛋白质和/或活性水平上诱导，尽管水平不同。然而，谷胱甘肽S-转移酶活性不被烯丙基、苯甲基和4-羟基苯甲基ITC诱导，谷胱甘肽还原酶活性不被苯甲基、4-羟基苯甲基ITC诱导，过氧化氢酶活性不被烯丙基ITC诱导。至于 Nrf2 转录因子，在所有测试的 ITC 处理 1 小时后，免疫印迹显示其蛋白质从胞质溶胶部分易位到细胞核。 ITC 诱导抗氧化剂和 II 相酶的能力似乎不受其亲水性或其他结构因素的影响。一起拍摄。这些结果表明，这些 ITC 是 ARE/Nrf2 调节的抗氧化/解毒基因的有效诱导剂，并且至少在大鼠肝脏中具有抑制依赖于 CYP3A2 催化的致癌物生物活性的潜力。 (C) 2012 Elsevier Ltd. 保留所有权利。
Primary cultures of rat hepatocytes were used to investigate whether and how eight isothiocynates (ITCs) with different chemical structures (the aromatic benzyl, 4-hydroxybenzyl, phenethyl isothiocyanates and the aliphatic ally!, napin, iberin, raphasatin isothiocyanates and sulforaphane) derived from hydrolyzed glucosinolates, were able to modulate cytochrome P450 (CYP) and antioxidant/detoxifying enzymes and to activate the Nrf2 transcription factor. The aromatic ITCs at 40 mu M markedly increased the transcription of CYP1A1 and 1A2 mRNA and increased the associated ethoxyresorufin O-deethylase (EROD) activity after 24 h of treatment. By contrast, the aliphatic ITCs (40 mu M) decreased CYP1A1 and 1A2 transcription, together with the corresponding EROD activity. The same treatment also caused a striking and similar transcriptional repression of CYP3A2, and the corresponding benzyloxyquinoline debenzylase activity in response to all the ITCs tested. In the same culture conditions, most of the antioxidant/detoxifying enzymes were significantly up-regulated by 40 mu M ITCs. In particular, NAD(P)H:quinone oxidoreductase and heme oxygenase-1 were induced, although to different levels, at transcriptional, protein and/or activity levels by all the ITCs. However, glutathione S-transferase activity was not induced by the allyl, benzyl, and 4-hydroxybenzyl ITCs, glutathione reductase activity was not induced by benzyl, and 4-hydroxybenzyl ITCs and catalase activity was not induced by allyl ITC. As for the Nrf2 transcription factor, a partial translocation of its protein from the cytosol to the nucleus was revealed by immunoblotting after 1 h of treatment for all the ITCs tested. The ability of ITCs to induce the antioxidant and phase II enzymes did not appear to be affected by their hydrophilicity or other structural factors. Taken together. these results show that these ITCs are effective inducers of ARE/Nrf2-regulated antioxidant/detoxifying genes and have the potential to inhibit, at least in rat liver, the bioactivation of carcinogens dependent on CYP3A2 catalysis. (C) 2012 Elsevier Ltd. All rights reserved.