Endothelial cell cytochrorne P450 1A1 and 1B1: Up-regulation by shear stress
Endothelial cell cytochrorne P450 1A1 and 1B1: Up-regulation by shear stress
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DOI：
10.1080/10623320490432434
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发表时间：
2004-01-01
影响因子：
--
通讯作者：
McIntire, LV
中科院分区：
文献类型：
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作者：
Eskin, SG;Turner, NA;McIntire, LV
将第三代人脐静脉内皮细胞（HUVEC）或第五代人主动脉内皮细胞（HAEC）在平行板流动系统中以25达因/cm（2）的速度处理24小时。将匹配的对照细胞保持在静态条件下。每次实验从6至8个载玻片中分离并合并总RNA。通过北方印迹和逆转录聚合酶链反应分析基因表达的变化。将倍数变化归一化为甘油醛磷酸脱氢酶（GAPDH）值。在HUVEC中，动脉水平的切应力使细胞色素P450 1A 1（CYP 1A 1）的mRNA表达增加10.8 +/- 2.1倍，CYP 1B 1增加23.1 +/- 3.7倍;而结缔组织生长因子（CTGF）的表达不变，内皮素-1（ET-1）的mRNA表达降低0.7 +/- 0.05倍。作者确定了这些变化是否是由β-萘黄酮（一种多环芳烃）诱导的，以及它们是否发生在HAEC中。β-萘酚酮上调HUVEC中的CYP 1A 1 18.3 +/- 4.2倍，和CYP 1B 1 4.1 +/- 0.3倍。剪切应力上调HAEC中的CYP 1A 1 6.3 +/- 0.4倍和CYP 1B 1 51.1 +/- 2.1倍。此外，作者还研究了从这些基因翻译的CYP 1A 1和CYP 1B 1蛋白。进行与上述相同的实验，收获细胞，通过CYP 1A 1和CYP 1B 1的蛋白质印迹进行蛋白质鉴定。切应力作用下，细胞色素P450 1A 1蛋白水平上调，而细胞色素P450 1B 1蛋白水平未上调。
Third-passage human umbilical vein endothelial cells (HUVECs) or fifth-passage human aortic endothelial cells (HAECs) were subjected to 25 dynes/cm(2) for 24 h in a parallel-plate flow system. Matched control cells were maintained in static conditions. Total RNA was isolated and pooled from six to eight slides per experiment. Changes in gene expression were analyzed by Northern blots and reverse transcriptase-polymerase chain reaction. Fold changes were normalized to glyceraldehyde phosphate dehydrogenase (GAPDH) values. In HUVECs, arterial levels of shear stress increased mRNA expression of Cytochrome P450 1A1 (CYP1A1) 10.8 +/- 2.1-fold, and CYP1B1 23.1 +/- 3.7-fold; whereas connective tissue growth factor (CTGF) expression was unchanged and endothelin-1 (ET-1) mRNA expression was decreased 0.7 +/- 0.05-fold. The authors determined whether these changes were induced by beta-naphthoflavone, a polyaromatic hydrocarbon, and whether they occurred in HAECs. beta-Naphthoflavone up-regulated CYP1A1 18.3 +/- 4.2-fold, and CYP1B1 4.1 +/- 0.3-fold in HUVECs. Shear stress up-regulated CYP1A1 6.3 +/- 0.4-fold and CYP1B1 51.1 +/- 2.1-fold in HAECs. In addition, the authors examined CYP1A1 and CYP1B1 proteins translated from these genes. Experiments identical to those described above were performed and the cells harvested for protein identification by Western blot of CYP1A1 and CYP1B1. Protein levels of CYP1A1 in HUVECs were up-regulated under shear stress, whereas protein levels of CYP1B1 were not.