2,3,7,8-tetrachlorodibenzo-p-dioxin increases reactive oxygen species production in human endothelial cells via induction of cytochrome P4501A1.
2,3,7,8-tetrachlorodibenzo-p-dioxin increases reactive oxygen species production in human endothelial cells via induction of cytochrome P4501A1.
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DOI：
10.1016/j.taap.2010.02.007
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发表时间：
2010-05-15
影响因子：
3.8
通讯作者：
Walker MK
中科院分区：
文献类型：
--
作者：
Kopf PG;Walker MK
我们实验室的研究表明，亚慢性暴露于成年小鼠的2，3，7，8，-四氯二苯并-对二恶英(TCDD)会导致高血压、心肌肥厚，并降低一氧化氮(NO)介导的血管扩张。此外，在暴露于TCDD的小鼠的心血管器官中观察到超氧阴离子的产生增加，这一增加有助于减少NO介导的血管扩张。由于细胞色素P4501A1(细胞色素P4501A1)可以参与TCDD诱导的某些毒性，我们验证了TCDD通过诱导细胞色素P4501A1增加内皮细胞内的ROS的假说。原代培养的人主动脉内皮细胞(HAECs)对TCDD暴露24 h(10 pm~10 nM)有浓度反应。氧化剂敏感荧光探针二氢乙二胺(DHE)和2‘，7’-二氯荧光素二乙酸酯(DCFH-DA)分别用于测定超氧阴离子、过氧化氢和羟基自由基。用二氨基荧光素-2-二乙酸酯(DAF-2DA)荧光探针测定NO含量。这些评估是在转染了针对芳香烃受体(AhR)、CYP1A1或CYP1B1的siRNA的HAEC中进行的。TCDD呈浓度依赖性地增加细胞色素P1A1和细胞色素P1B1的mRNA、蛋白和酶活性。1 nM TCDD使DHE(CONT=1.0±0.3；TCDD=5.1±1.0；p=0.002)和DCFH-DA(CONT=1.0±0.2；TCDD=4.1±0.5；p=0.002)荧光增强，DAF-2DA荧光最大幅度降低(CONT=1.0±0.4；TCDD=0.68±0.1)。针对AhR和CyP1A1的siRNA显著降低TCDD诱导的DHE(siAhR：cont=1.0±0.1；tCDD=1.3±0.2；p=0.093)(siAhR：cont=1.0±0.1；tCDD=1.1±0.1；p=0.454)和DCFH-DA(siAhR：cont=1.0±0.2；tCDD=1.3±0.3；p=0.370)(siCyP1A1：cont=1.0±0.1；tCDD=1.3±0.2；p=0.114)，并增加DAF-2DA荧光(AhsiR：cont=1.00±0.03；tCDD=0.97±0.03；p=0.481)(siAhR：cont=1.0±0.1；tCDD=1.3±0.2；p=0.114)TCDD=0.92±0.03；p=0.034)，而靶向细胞色素P1B1的siRNA则无此作用。这些数据表明，TCDD诱导的ROS增加是AhR依赖的，可能部分是由CYP1A1诱导的。
Studies in our laboratory have demonstrated that subchronic 2,3,7,8,-tetrachlorodibenzo-p-dioxin (TCDD) exposure of adult mice results in hypertension, cardiac hypertrophy, and reduced nitric oxide (NO)-mediated vasodilation. Moreover, increased superoxide anion production was observed in cardiovascular organs of TCDD-exposed mice and this increase contributed to the reduced NO-mediated vasodilation. Since cytochrome P4501A1 (CYP1A1) can contribute to some TCDD-induced toxicity, we tested the hypothesis that TCDD increases reactive oxygen species (ROS) in endothelial cells by the induction of CYP1A1. A concentration-response to 24 h TCDD exposure (10 pM-10 nM) was performed in confluent primary human aortic endothelial cells (HAECs). Oxidant-sensitive fluorescent probes dihydroethidium (DHE) and 2’,7’-dichlorofluorescin diacetate (DCFH-DA), were used to measure superoxide anion, and hydrogen peroxide and hydroxyl radical, respectively. NO was also measured using the fluorescent probe diaminofluorescein-2 diacetate (DAF-2DA). These assessments were conducted in HAECs transfected with siRNA targeting the aryl hydrocarbon receptor (AhR), CYP1A1, or CYP1B1. TCDD concentration-dependently increased CYP1A1 and CYP1B1 mRNA, protein, and enzyme activity. Moreover, 1 nM TCDD maximally increased DHE (Cont=1.0±0.3; TCDD=5.1±1.0; p=0.002) and DCFH-DA (Cont=1.0±0.2; TCDD=4.1±0.5; p=0.002) fluorescence and maximally decreased DAF-2DA fluorescence (Cont=1.0±0.4; TCDD=0.68±0.1). siRNA targeting AhR and CYP1A1 significantly decreased TCDD-induced DHE (siAhR: Cont=1.0±0.1; TCDD=1.3±0.2; p=0.093) (siCYP1A1: Cont=1.0±0.1; TCDD=1.1±0.1; p=0.454) and DCFH-DA (siAhR: Cont=1.0±0.2; TCDD=1.3±0.3; p=0.370) (siCYP1A1: Cont=1.0±0.1; TCDD=1.3±0.2; p=0.114) fluorescence and increased DAF-2DA fluorescence (siAhR: Cont=1.00±0.03; TCDD=0.97±0.03; p=0.481) (siCYP1A1: Cont=1.00±0.03; TCDD=0.92±0.03; p=0.034), while siRNA targeting CYP1B1 did not. These data suggest that TCDD-induced increase in ROS is AhR dependent and may be mediated, in part, by CYP1A1 induction.
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影响因子：
4.5
作者：
Garrick, RA;Woodin, BR;Stegeman, JJ
通讯作者：
Stegeman, JJ
影响因子：
3.8
作者：
Dong, W;Teraoka, H;Hiraga, T
通讯作者：
Hiraga, T
影响因子：
3.8
作者：
Hornug, MW;Spitsbergen, JM;Peterson, RE
通讯作者：
Peterson, RE
影响因子：
3.8
作者：
Guiney, PD;Smolowitz, RM;Stegeman, JJ
通讯作者：
Stegeman, JJ
DOI：
10.1080/10623320490432434
发表时间：
2004-01-01
影响因子：
--
作者：
Eskin, SG;Turner, NA;McIntire, LV
通讯作者：
McIntire, LV