CYCLIC STRAIN UP-REGULATES NITRIC-OXIDE SYNTHASE IN CULTURED BOVINE AORTIC ENDOTHELIAL-CELLS
CYCLIC STRAIN UP-REGULATES NITRIC-OXIDE SYNTHASE IN CULTURED BOVINE AORTIC ENDOTHELIAL-CELLS
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DOI：
10.1172/jci118181
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发表时间：
1995-09-01
影响因子：
15.9
通讯作者：
SUMPIO, BE
中科院分区：
文献类型：
--
作者：
AWOLESI, MA;SESSA, WC;SUMPIO, BE
在体内，内皮细胞（EC）受到血液动力学的影响，这可能会影响一氧化氮的产生。本研究旨在观察周期性应变对培养的牛主动脉EC内皮型一氧化氮合酶（eNOS）表达的影响。EC在柔性膜上生长，所述柔性膜在-5或-20kPa的真空下以60个循环/分钟经受变形。这导致平均应变分别为6和10%，其被传递到附着的细胞。总胞质RNA的北方印迹分析表明，两种应变方案均增加了eNOS基因表达，但10%平均应变的增加大于6%平均应变的增加。核径流转录测定证实了eNOS转录本的诱导。Western blot分析显示，与对照组或6%平均应变相比，10%平均应变24小时后eNOS水平增加。在-20 kPa真空下变形的膜的高应变周边（7-24%应变）中，EC对eNOS的免疫组织化学染色增加。这些结果表明，周期性应变上调eNOS的转录本和蛋白质水平的表达在牛主动脉EC，从而强调在体内的eNOS调节的血液动力学力量的重要性。
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.