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首页> 外文期刊>Endothelium: Journal of endothelial cell research >Endothelial cell cytochrome P450 1A1 and 1B1: up-regulation by shear stress.
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Endothelial cell cytochrome P450 1A1 and 1B1: up-regulation by shear stress.

机译：内皮细胞细胞色素P450 1A1和1B1：剪切应力上调。

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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.
机译：在平行板流动系统中，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）经受25达因/厘米（2）的作用24 h。匹配的对照细胞保持在静态条件下。分离总RNA，并在每个实验中合并六到八个玻片。通过Northern印迹和逆转录酶-聚合酶链反应分析基因表达的变化。将倍数变化标准化为甘油醛磷酸脱氢酶（GAPDH）值。在HUVEC中，动脉水平的切应力增加了细胞色素P450 1A1（CYP1A1）的mRNA表达10.8 +/- 2.1倍和CYP1B1 23.1 +/- 3.7倍;而结缔组织生长因子（CTGF）的表达保持不变，而内皮素1（ET-1）的mRNA表达则下降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。在HUVEC中，β-萘黄酮上调CYP1A1 18.3 +/- 4.2倍，而CYP1B1 4.1 +/- 0.3倍。在HAEC中，剪切应力上调CYP1A1 6.3 +/- 0.4倍和CYP1B1 51.1 +/- 2.1倍。此外，作者检查了从这些基因翻译的CYP1A1和CYP1B1蛋白。进行与上述相同的实验，并收集细胞以通过CYP1A1和CYP1B1的蛋白质印迹进行蛋白质鉴定。在剪应力作用下，HUVECs中的CYP1A1蛋白质水平上调，而CYP1B1的蛋白质水平则没有上调。

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