Soy isoflavones increase quinone reductase in hepa-1c1c7 cells via estrogen receptor beta and nuclear factor erythroid 2-related factor 2 binding to the antioxidant response element
Soy isoflavones increase quinone reductase in hepa-1c1c7 cells via estrogen receptor beta and nuclear factor erythroid 2-related factor 2 binding to the antioxidant response element
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DOI：
10.1016/j.jnutbio.2010.07.008
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发表时间：
2011-09-01
影响因子：
5.6
通讯作者：
Steinberg, Francene M.
中科院分区：
文献类型：
--
作者：
Froyen, Erik B.;Steinberg, Francene M.
在动物和细胞培养模型中，大豆蛋白和大豆黄酮（染料木素和大豆苷元）已被证明可增加醌还原酶（QR）活性、蛋白质和mRNA。然而，其作用机制尚未完全确定。此外，尚未确定雌马酚（大豆苷元代谢物）是否可以调节QR活性和表达。雌激素受体β（ER β）被认为与核因子红细胞2相关因子2（Nrf 2）沿着通过抗雌激素和植物雌激素参与刺激QR基因转录。本研究验证了染料木素、大豆苷元和雌马酚通过ER β和Nrf 2与QR抗氧化反应元件（ARE）结合来增加醌还原酶活性、蛋白质和mRNA的假设。使用TaqMan聚合酶链反应、蛋白质免疫印迹和活性测定来确定QR表达和活性。使用荧光素酶报告基因测定和染色质免疫沉淀（ChIP）研究分子事件。Hepa-1c 1c 7细胞用对照[0.1%（v：v）二甲亚砜（DMSO）]、1 μ mol/L β-萘酮（阳性对照）、5 μ mol/L白藜芦醇（用于ER β结合的ChIP阳性对照）和1、5和25 μ mol/L染料木黄酮、大豆苷元或雌马酚处理。处理持续时间为1 h（ChIP）、24 h（mRNA和荧光素酶测定）以及24和48 h（蛋白质和活性）。染料木黄酮，大豆黄酮和雌马酚增加QR活性，蛋白质和mRNA，大豆黄酮和雌马酚有更多的影响，在生理浓度（1和5 μ mol/L）相比，染料木黄酮。此外，研究结果表明，染料木素，大豆苷元和雌马酚与QR ARE相互作用，大豆苷元和雌马酚通过ER β和Nrf 2强烈结合QR ARE发挥作用。(C)2011 Elsevier Inc. All rights reserved.
Soy protein and isoflavones (genistein and daidzein) have been demonstrated to increase quinone reductase (QR) activity, protein, and mRNA in animal and cell culture models. However, their mechanism of action has not been completely characterized. Additionally, it has not been determined if equol, a daidzein metabolite, can modulate QR activity and expression. Estrogen receptor beta (ER beta) is thought to be involved in stimulating QR gene transcription by anti-estrogens and phytoestrogens, along with nuclear factor erythroid 2-related factor 2 (Nrf2). This study tested the hypothesis that genistein, daidzein and equol increase quinone reductase activity, protein and mRNA via ER beta and Nrf2 binding to the QR antioxidant response element (ARE). QR expression and activity were determined using TaqMan polymerase chain reaction, protein immunoblots and activity assays. Molecular events were investigated using luciferase reporter gene assays and chromatin immunoprecipitation (ChIP). Hepa-1c1c7 cells were treated with control [0.1% (v:v) dimethyl sulfoxide (DMSO)]; 1 mu mol/L beta-naphthoflavone (Positive control); 5 mu mol/L resveratrol (ChIP positive control for ER beta binding) and 1,5 and 25 mu mol/L genistein, daidzein or equol. Treatment durations were 1 h (ChIP), 24 h (mRNA and luciferase assays) and 24 and 48 h (protein and activity). Genistein, daidzein and equol increased QR activity, protein and mRNA, with daidzein and equol having more of an impact at physiologic concentrations (1 and 5 mu mol/L) compared to genistein. Furthermore, the study results demonstrate that genistein, daidzein and equol interact with the QR ARE and that daidzein and equol act via both ER beta and Nrf2 binding strongly to the QR ARE. (C) 2011 Elsevier Inc. All rights reserved.