Regulation of MUC5AC expression by NAD(P)H:quinone oxidoreductase 1
Regulation of MUC5AC expression by NAD(P)H:quinone oxidoreductase 1
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DOI：
10.1016/j.freeradbiomed.2007.01.040
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发表时间：
2007-05-01
影响因子：
7.4
通讯作者：
Voynow, Judith A.
中科院分区：
文献类型：
--
作者：
Zheng, Shuo;Byrd, Angela S.;Voynow, Judith A.
中性粒细胞弹性蛋白酶 (NE) 是一种有效的中性粒细胞炎症介质，通过涉及活性氧的未定义途径增加 MUC5AC 粘蛋白基因表达。为了确定 NE 产生的活性氧的来源，我们使用氧化还原酶的药理学抑制剂来测试它们是否阻断 NE 调节的 MUC5AC mRNA 表达。我们发现双香豆素是 NADP(H)：醌氧化还原酶 1 (NQO1) 的抑制剂，可抑制 A549 肺腺癌细胞和原代正常人支气管上皮细胞中 MUC5AC mRNA 的表达。我们通过使用短干扰RNA（siRNA）抑制NQO1表达进一步测试了NQO1在介导NE诱导的MUC5AC表达中的作用。用 NQO1 特异性 siRNA 转染可抑制 NQO1 表达并显着消除 MUC5AC mRNA 表达。 NE处理引起A549细胞脂质过氧化；这种效应被双香豆素预处理所抑制，表明 NQO1 还可以调节 NE 暴露后 A549 细胞中的氧化应激。 NE 暴露增加了 NQO1 蛋白和活性水平； NQO1 的表达和活性仅限于细胞质，不会转移到质膜。我们的结果表明，NQO1 作为 NE 调节的氧化应激和 MUC5AC 粘蛋白基因表达的关键介质发挥着重要作用。 (c) 2007 Elsevier Inc. 保留所有权利。
Neutrophil elastase (NE), a potent neutrophil inflammatory mediator, increases MUC5AC mucin gene expression through undefined pathways involving reactive oxygen species. To determine the source of NE-generated reactive oxygen species, we used pharmacologic inhibitors of oxidoreductases to test whether they blocked NE-regulated MUC5AC mRNA expression. We found that dicumarol, an inhibitor of the NADP(H): quinone oxidoreductase 1 (NQO1), inhibited MUC5AC mRNA expression in A549 lung adenocarcinoma cells and primary normal human bronchial epithelial cells. We further tested the role of NQO1 in mediating NE-induced MUC5AC expression by inhibiting NQO1 expression using short interfering RNA (siRNA). Transfection with siRNA specific for NQO1 suppressed NQO1 expression and significantly abrogated MUC5AC mRNA expression. NE treatment caused lipid peroxidation in A549 cells; this effect was inhibited by pretreatment with dicumarol, suggesting that NQO1 also regulates oxidant stress in A549 cells after NE exposure. NE exposure increased NQO1 protein and activity levels; NQO1 expression and activity were limited to the cytosol and did not translocate to the plasma membrane. Our results indicate that NQO1 has an important role as a key mediator of NE-regulated oxidant stress and MUC5AC mucin gene expression. (c) 2007 Elsevier Inc. All rights reserved.