首页> 美国卫生研究院文献>The Journal of Clinical Investigation >Cyclic strain upregulates nitric oxide synthase in cultured bovine aortic endothelial cells.
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Cyclic strain upregulates nitric oxide synthase in cultured bovine aortic endothelial cells.

机译:循环菌株在培养的牛主动脉内皮细胞中上调一氧化氮合酶。

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摘要

In vivo, endothelial cells (EC) are subjected to hemodynamic forces which may influence the production of nitric oxide. This study was designed to examine the effect of cyclic strain on the expression of endothelial nitric oxide synthase (eNOS) in cultured bovine aortic EC. EC were grown on flexible membranes which were subjected to deformation at 60 cycles/min with -5 or -20 kPa of vacuum. This results in an average strain of 6 and 10%, respectively, which is transmitted to the attached cells. Northern blot analysis of total cytosolic RNA demonstrated an increase in eNOS gene expression with both strain regimens but the increase with 10% average strain was greater than that at 6%. Nuclear runoff transcription assays confirmed the induction of eNOS transcripts. Western blot analysis showed an increase in eNOS level after 24 h of cyclic 10% average strain compared with controls or 6% average strain. Immunohistochemical staining of EC for eNOS was increased in the high strain periphery (7-24% strain) of membranes deformed with -20 kPa vacuum. These results demonstrate that cyclic strain upregulates the expression of eNOS transcripts and protein levels in bovine aortic EC thus emphasizing the importance of hemodynamic forces in the regulation of eNOS in vivo.
机译:在体内,内皮细胞(EC)受到血液动力的作用,这可能会影响一氧化氮的产生。这项研究旨在检查循环应变对培养的牛主动脉EC中内皮型一氧化氮合酶(eNOS)表达的影响。 EC在柔性膜上生长,该柔性膜在-5或-20kPa的真空下以60个循环/分钟的速度变形。这导致平均应变分别为6%和10%,并传递到附着的细胞。对总胞质RNA的Northern印迹分析表明,两种菌株方案中eNOS基因表达均增加,但平均菌株为10%时的增加大于6%时。核径流转录测定法证实了eNOS转录物的诱导。 Western印迹分析显示,与对照组或6%平均菌株相比,循环10%平均菌株24 h后eNOS水平升高。 EC的eNOS免疫组织化学染色在-20 kPa真空下变形的高应变膜周边(7-24%应变)增加。这些结果表明,循环应变上调了牛主动脉EC中eNOS转录物的表达和蛋白质水平,从而强调了血流动力学力在体内eNOS调控中的重要性。

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