Cattaruzza Marco, Guzik Tomasz J, Słodowski Wojciech, Pelvan Ayşegül, Becker Jürgen, Halle Martin, Buchwald Arnd B, Channon Keith M, Hecker Markus
Department of Cardiovascular Physiology, University of Göttingen, Göttingen, Germany.
Circ Res. 2004 Oct 15;95(8):841-7. doi: 10.1161/01.RES.0000145359.47708.2f. Epub 2004 Sep 16.
Coronary heart disease (CHD) is based on the development of atherosclerosis in coronary arteries. Shear stress-induced endothelial nitric oxide (NO) release not only contributes to local blood pressure control but also effectively helps to retard atherosclerosis. Therefore, functionally relevant polymorphisms in the endothelial NO synthase (NOS-3) gene may contribute to the development of CHD. NOS-3 expression was analyzed in endothelial cells isolated from umbilical cords genotyped for the -786C/T single nucleotide polymorphism (SNP) of the human nos-3 gene. Moreover, NO-dependent relaxation was examined in segments of saphenous vein isolated from genotyped patients undergoing aortocoronary bypass surgery, and patients subjected to quantitative coronary angiography were genotyped to verify an association between this SNP and CHD. Shear stress-induced NOS-3 mRNA and protein expression was present in TT and CT genotype cells but absent in cells with CC genotype. Pretreatment of these cells with a decoy oligonucleotide comprising position -800 to -779 of the C-type nos-3 promoter reconstituted shear stress-induced NOS-3 expression. These results were confirmed by reporter gene analysis with the corresponding nos-3 promoter luciferase constructs. In addition, the NO-mediated relaxant response of vein grafts from CC genotype patients was significantly attenuated as compared with the CT or TT genotype, and in CHD-positive patients, the CC genotype was significantly more frequent (19.0%) than in CHD-negative patients (4.4%). The -786C/T SNP of the nos-3 gene thus constitutes a genetic risk factor for CHD, presumably due to binding of an inhibitory transcription factor to the C-type promoter blocking shear stress-dependent maintenance of NOS-3 expression.
冠心病（CHD）是基于冠状动脉粥样硬化的发展。剪切应力诱导的内皮型一氧化氮（NO）释放不仅有助于局部血压控制，还能有效帮助延缓动脉粥样硬化。因此，内皮型一氧化氮合酶（NOS-3）基因中功能相关的多态性可能促成冠心病的发展。对从脐带分离的内皮细胞进行分析，这些脐带针对人类nos-3基因的-786C/T单核苷酸多态性（SNP）进行了基因分型。此外，在接受主动脉冠状动脉搭桥手术的基因分型患者分离的大隐静脉段中检测了NO依赖性舒张，并且对接受定量冠状动脉造影的患者进行基因分型以验证该SNP与冠心病之间的关联。剪切应力诱导的NOS-3 mRNA和蛋白表达在TT和CT基因型细胞中存在，但在CC基因型细胞中不存在。用包含C型nos-3启动子-800至-779位置的诱饵寡核苷酸预处理这些细胞可重建剪切应力诱导的NOS-3表达。这些结果通过使用相应的nos-3启动子荧光素酶构建体的报告基因分析得到证实。此外，与CT或TT基因型相比，CC基因型患者静脉移植物的NO介导的舒张反应明显减弱，并且在冠心病阳性患者中，CC基因型的频率（19.0%）明显高于冠心病阴性患者（4.4%）。因此，nos-3基因的-786C/T SNP构成冠心病的遗传危险因素，可能是由于抑制性转录因子与C型启动子结合，阻断了剪切应力依赖性的NOS-3表达维持。
