Reduced gene expression of vascular endothelial NO synthase and cyclooxygenase-1 in heart failure.
Reduced gene expression of vascular endothelial NO synthase and cyclooxygenase-1 in heart failure.
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DOI：
10.1161/01.res.78.1.58
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发表时间：
1996
影响因子：
20.1
通讯作者：
Carolyn J. Smith;Dong Sun;C. Hoegler;Barrie S. Roth;Xiaoping Zhang;Gong Zhao;Xiaobin Xu;Y. Kobari
中科院分区：
文献类型：
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作者：
Carolyn J. Smith;Dong Sun;C. Hoegler;Barrie S. Roth;Xiaoping Zhang;Gong Zhao;Xiaobin Xu;Y. Kobari
在犬起搏诱导的心力衰竭和人类各种病因的心力衰竭发作后，冠状动脉和外周血管中的内皮依赖性反应受到抑制。本研究旨在探讨这些反应是否是由于内皮细胞一氧化氮合酶（ecNOS）和环氧合酶-1（考克斯-1）的表达减少。在左心室起搏1个月后，监测8只杂种犬的心力衰竭，心力衰竭由临床体征和左心室舒张末期压> 25 mm Hg定义。从从胸主动脉刮取的内皮细胞中分离总RNA和蛋白质，并分别通过北方和Western印迹分析。用32 P标记的ecNOS和考克斯-1 cDNA探针印迹，通过光密度定量，并将结果相对于GAPDH或von Willebrand因子（vWF）标准化。在任意单位中，心衰前后ecNOS与GAPDH的比值分别为2.66 +/- 0.77（平均值+/- SEM，n = 17）和1.12 +/- 0.37（n = 6），考克斯-1与GAPDH的比值分别为1.52 +/- 0.52和0.56 +/- 0.15。这些代表ecNOS和考克斯-1基因表达降低56%至64%（P <0.05）。考克斯-1或ecNOS与vWF的比值无变化。心力衰竭后ecNOS蛋白也明显减少，估计为70%。一个显着的减少亚硝酸盐的生产，酶活性的措施，从胸部aerotas在响应刺激的乙酰胆碱或缓激肽也发生。为了确定ecNOS和考克斯-1是否可以独立调节，口服活性NO释放剂CAS 936给7只正常狗7天，并分析主动脉ecNOS和考克斯-1 mRNA。在这些狗的子宫内膜中，ecNOS与GAPDH的比率降低了52%（P <0.05），而考克斯-1与GAPDH的比率没有变化。当数据标准化为vWF时，发现了类似的结果。这些结果表明，至少有两个内皮血管扩张基因产物减少心力衰竭，而不是选择性缺陷的NO合酶基因表达。
Endothelium-dependent responses are depressed in coronary and peripheral blood vessels after the onset of pacing-induced heart failure in dogs and heart failure of various etiologies in humans. The present study was designed to examine whether these responses were due to decreases in the expression of endothelial cell NO synthase (ecNOS) and cyclooxygenase-1 (COX-1). After 1 month of left ventricular pacing, 8 mongrel dogs were monitored for heart failure as defined by clinical signs and left ventricular end diastolic pressures > 25 mm Hg. Total RNA and protein were isolated from endothelial cells scraped from the thoracic aorta and analyzed by Northern and Western blotting, respectively. Blots probed with 32P-labeled cDNAs for ecNOS and COX-1 were quantified densitometrically, and results were normalized against GAPDH or von Willebrand factor (vWF). In arbitrary units, the ratios of ecNOS to GAPDH were 2.66 +/- 0.77 (mean +/- SEM, n = 17) and 1.12 +/- 0.37 (n = 6 and the ratios of COX-1 to GAPDH were 1.52 +/- 0.52 and 0.56 +/- 0.15 before and after heart failure, respectively. These represent 56% to 64% (P < .05) reductions in ecNOS and COX-1 gene expression. There was no change in the ratios of either COX-1 or ecNOS to vWF. There was also a marked reduction in ecNOS protein after heart failure, estimated at 70%. A marked reduction in nitrite production, a measure of enzyme activity, from thoracic aortas in response to stimulation by either acetylcholine or bradykinin also occurred. To determine whether ecNOS and COX-1 could be independently regulated, an orally active NO-releasing agent, CAS 936, was given to 7 normal dogs for 7 days, and aortic ecNOS and COX-1 mRNAs were analyzed. The ratio of ecNOS to GAPDH was depressed by 52% (P < .05) in aortas from these dogs, whereas the ratio of COX-1 to GAPDH was unchanged. Similar results were found when data were normalized to vWF. These results suggest that at least two endothelial vasodilator gene products are reduced in heart failure, as opposed to a selective defect in NO synthase gene expression.