Davies M G, Berkowitz D E, Hagen P O
Department of Surgery, Duke University Medical Center, Durham, North Carolina 27710, USA.
J Surg Res. 1995 Jun;58(6):732-8. doi: 10.1006/jsre.1995.1116.
Although little is known about the endothelial cell function of human saphenous vein coronary artery bypass grafts, there is evidence to suggest that receptor-activated, endothelial-dependent relaxation mediated by nitric oxide is impaired. This study examines the expression and function of endothelial cell constitutive nitric oxide synthase (cNOS) of aortocoronary vein bypass grafts and human saphenous veins obtained from 10 patients undergoing repeat coronary artery bypass grafting for recurrent ischemic symptoms. Following precontraction with norepinephrine (10(-5) M), responses to acetylcholine (receptor-mediated, endothelium-dependent), calcium ionophore (A23187; receptor-independent, endothelium-dependent), and sodium nitroprusside (endothelium-independent) were assessed. Following total RNA extraction using phenol/guanidinium isothiocyanate from specimens of human saphenous vein and vein graft, a quantitative RNase Protection Assay (RPA) was performed using a cRNA riboprobe corresponding to a fragment of the human endothelial cell cNOS gene. Histologically, the vein grafts showed both intimal hyperplasia development and focal atherosclerosis formation compared to the saphenous veins. Scanning electron microscopy of the saphenous veins and the vein grafts showed an intact endothelium. Precontracted vein grafts did not relax in response to acetylcholine; in contrast, the saphenous vein relaxed in a dose-dependent manner to reach a maximal relaxation of 19 +/- 4% precontracted tension. Saphenous veins and vein grafts relaxed in response to A23187 with maximal relaxation of 92 +/- 5 and 73 +/- 13%, respectively. Both vessels relaxed in a dose dependent manner to sodium nitroprusside. RPA normalized to beta-actin showed similar levels of expression of endothelial cell cNOS equivalent to 1 pg of sense RNA in both the saphenous vein and vein graft.(ABSTRACT TRUNCATED AT 250 WORDS)
尽管人们对人体大隐静脉冠状动脉旁路移植血管的内皮细胞功能了解甚少,但有证据表明,由一氧化氮介导的受体激活的内皮依赖性舒张功能受损。本研究检测了取自10例因复发性缺血症状接受再次冠状动脉旁路移植术患者的主动脉冠状动脉静脉旁路移植血管和人体大隐静脉中内皮细胞组成型一氧化氮合酶(cNOS)的表达及功能。用去甲肾上腺素(10⁻⁵ M)预收缩后,评估对乙酰胆碱(受体介导的内皮依赖性)、钙离子载体(A23187;非受体介导的内皮依赖性)和硝普钠(非内皮依赖性)的反应。从人体大隐静脉和静脉移植血管标本中用苯酚/异硫氰酸胍提取总RNA后,使用与人内皮细胞cNOS基因片段对应的cRNA核糖探针进行定量核糖核酸酶保护分析(RPA)。组织学上,与大隐静脉相比,静脉移植血管显示出内膜增生和局灶性动脉粥样硬化形成。大隐静脉和静脉移植血管的扫描电子显微镜检查显示内皮完整。预收缩的静脉移植血管对乙酰胆碱无舒张反应;相反,大隐静脉以剂量依赖性方式舒张,达到预收缩张力的最大舒张为19±4%。大隐静脉和静脉移植血管对A23187有舒张反应,最大舒张分别为92±5%和73±13%。两种血管对硝普钠均呈剂量依赖性舒张。以β-肌动蛋白标准化的RPA显示,大隐静脉和静脉移植血管中内皮细胞cNOS的表达水平相似,相当于1 pg有义RNA。(摘要截短至250字)