Reciprocal regulation of endothelin-1 and endothelial constitutive NOS in proliferating endothelial cells
Reciprocal regulation of endothelin-1 and endothelial constitutive NOS in proliferating endothelial cells
复制标题
DOI：
10.1152/ajpheart.1995.269.6.h1988
复制
发表时间：
1995-12-01
影响因子：
4.8
通讯作者：
Marsden, PA
中科院分区：
文献类型：
--
作者：
Flowers, MA;Wang, Y;Marsden, PA
内皮素-1 （ET-1）和内皮型一氧化氮合酶（ecNOS）的表达在两种独立的体外模型中进行了评估：异步差异增殖培养和损伤内皮细胞单层。对融合前、融合和融合后细胞分离的RNA进行Northern blot分析发现，增殖细胞中ET-1 mRNA转录物水平增加了四倍，而ecNOS mRNA转录物水平降低了两倍。核径流分析表明，增殖细胞中稳态ET-1 mRNA含量的增加部分是由基因转录增加介导的。与静止非增殖细胞相比，增殖细胞中ecNOS转录率没有降低，表明mRNA不稳定介导了增殖内皮中ecNOS mRNA水平的降低。ET-1和ecNOS的蛋白表达变化一致。在受损的内皮细胞单层中，原位cRNA杂交显示，受损内皮单层生长前沿的ET-1 mRNA转录水平升高。这些数据表明ET-1和ecNOS的表达在两种不同的内皮细胞增殖模型中相互调节。
The expression of endothelin-1 (ET-1) and endothelial constitutive nitric oxide synthase (ecNOS) was assessed in two independent in vitro models: asynchronously differentially proliferating cultures and wounded endothelial cell monolayers. Northern blot analysis of RNA isolated from preconfluent, confluent, and postconfluent cells revealed a fourfold rise in ET-1 mRNA transcripts, whereas levels of ecNOS mRNA transcripts were reduced twofold in proliferating cells. Nuclear run-off analysis demonstrated that increased steady-state ET-1 mRNA content in proliferating cells was mediated, in part, by increased gene transcription. In contrast, ecNOS transcription rates in proliferating cells were not decreased compared with quiescent nonproliferating cells, indicating that mRNA destabilization mediated the decreased ecNOS mRNA levels in proliferating endothelium. Concordant changes in protein expression were documented for both ET-1 and ecNOS. In injured endothelial cell monolayers, in situ cRNA hybridization demonstrated increased mRNA transcript levels for ET-1 in growth fronts of injured endothelial monolayers. These data are taken to indicate that expression of ET-1 and ecNOS is reciprocally regulated in two different models of endothelial cell proliferation.