Superoxide-mediated inactivation of nitric oxide and peroxynitrite formation by tobacco smoke in vascular endothelium: studies in cultured cells and smokers
Superoxide-mediated inactivation of nitric oxide and peroxynitrite formation by tobacco smoke in vascular endothelium: studies in cultured cells and smokers
复制标题
DOI：
10.1152/ajpheart.00930.2008
复制
发表时间：
2009-06-01
影响因子：
4.8
通讯作者：
Radi, Rafael
中科院分区：
文献类型：
--
作者：
Peluffo, Gonzalo;Calcerrada, Pablo;Radi, Rafael
张文辉，张文辉.烟草烟雾对血管内皮细胞中一氧化氮和过氧亚硝酸盐形成的超氧化物介导的失活：对培养细胞和吸烟者的研究。Am J Physiol Heart Circ Physiol 296：H1781-H1792，2009.首次发表于2009年4月10日; doi：10.1152/ajpheart.00930.2008。已知烟草烟雾会导致一氧化氮（（NO）-N-中心点）失活和内皮功能障碍。在这项工作中，我们评估了（NO）-N-中心点和超氧（O-2（中心点-））自由基之间的相互作用，以及随后的影响（NO）-N-中心点的生物利用度和氮氧化应激暴露于香烟烟雾提取物（CSE）的牛主动脉内皮细胞和吸烟者。如自旋捕获电子顺磁共振实验所示，在CSE存在下牛主动脉内皮细胞触发O-2（中心点-）产生。细胞外（3.4与1.0 nmol.h（-1）.mg（-1）; CSE与对照;细胞色素c（3+）还原试验）和细胞内（40%细胞溶质乌头酸酶抑制）均产生O-2（中心点-）。CSE还导致过亚硝酸盐的产生，如通过细胞上的二氢罗丹明氧化和蛋白质酪氨酸硝化所评估的。O-2（中心点-）和过氧亚硝酸盐的形成减少抗坏血酸和α-生育酚。此外，CSE导致内皮型一氧化氮合酶的氧化，增加内皮型一氧化氮合酶的单体失活形式。吸烟者和年龄匹配的健康志愿者口服补充500毫克抗坏血酸加400国际单位全消旋α-生育酚每12小时165天。通过血流介导的肱动脉扩张评估，吸烟者与对照组相比有内皮功能障碍（95%置信区间：2.5，8.3 vs. 10.6，14.2; P < 0.05），血浆蛋白3-硝基酪氨酸水平高1.4倍。吸烟者的血流介导的舒张功能丧失在长期补充抗氧化剂后恢复（95%置信区间：13.9，19.9; P < 0.05），达到与对照人群相当的值。我们的数据表明，烟草烟雾中的元素，最有可能通过氧化还原循环，转向（NO）-N-中心点对过氧亚硝酸盐诱导O-2（中心点-）生产在血管内皮细胞在体外和体内。
Peluffo G, Calcerrada P, Piacenza L, Pizzano N, Radi R. Superoxide-mediated inactivation of nitric oxide and peroxynitrite formation by tobacco smoke in vascular endothelium: studies in cultured cells and smokers. Am J Physiol Heart Circ Physiol 296: H1781-H1792, 2009. First published April 10, 2009; doi:10.1152/ajpheart.00930.2008.-Tobacco smoke is known to cause nitric oxide ((NO)-N-center dot) inactivation and endothelial dysfunction. In this work we evaluated the interplay between (NO)-N-center dot and superoxide (O-2(center dot-)) radicals and the consequent impact on (NO)-N-center dot bioavailability and nitroxidative stress in bovine aortic endothelial cells exposed to cigarette smoke extract (CSE) and in smokers. Bovine aortic endothelial cells in the presence of CSE triggered O-2(center dot-) production as indicated by spin-trapping electron paramagnetic resonance experiments. O-2(center dot-) was produced both extracellulary (3.4 vs. 1.0 nmol.h(-1).mg(-1); CSE vs. control; cytochrome c(3+) reduction assay) and intracellularly (40% inhibition of cytosolic aconitase). CSE also led to the production of peroxynitrite as evaluated by dihydrorhodamine oxidation and protein tyrosine nitration on cells. O-2(center dot-) and peroxynitrite formation were decreased by ascorbate and alpha-tocopherol. Additionally, CSE led to the oxidation of endothelial nitric oxide synthase increasing the monomeric inactive form of endothelial nitric oxide synthase. Smokers and age-matched healthy volunteers were supplemented orally with 500 mg ascorbate plus 400 IU all-rac-alpha-tocopherol every 12 h for 165 days. Smokers had endothelial dysfunction compared with control subjects (95% confidence interval: 2.5, 8.3 vs. 10.6, 14.2; P < 0.05) as assessed by flow-mediated dilation of the brachial artery, and plasma levels of protein 3-nitrotyrosine were 1.4-fold higher. The loss of flow-mediated dilation in smokers reverted after a long- term antioxidant supplementation (95% confidence interval: 13.9, 19.9; P < 0.05), reaching values comparable with the control population. Our data indicate that elements on tobacco smoke, most likely through redox cycling, divert (NO)-N-center dot toward peroxynitrite by inducing O-2(center dot-) production in vascular endothelial cells both in vitro and in vivo.