Formation of nitric oxide, superoxide, and peroxynitrite in myocardial ischemia-reperfusion injury in rats
Formation of nitric oxide, superoxide, and peroxynitrite in myocardial ischemia-reperfusion injury in rats
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DOI：
10.1152/ajpheart.1997.272.5.h2327
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发表时间：
1997-05-01
影响因子：
4.8
通讯作者：
Wong, PYK
中科院分区：
文献类型：
--
作者：
Liu, PT;Hock, CE;Wong, PYK
在本研究中，一氧化氮（NO），超氧化物和过氧亚硝酸盐的心肌缺血-再灌注（MI/R）诱导的炎症反应的贡献进行了研究。将雄性Sprague-Dawley大鼠麻醉，结扎左冠状动脉主干20 min，再灌注5 h。MI/R诱导严重心律失常，表现为MI/R组心律失常评分显著高于假手术对照组。MI/R组左心室游离壁肌酸激酶活性显著降低38%。相反，MI/R组左心室游离壁髓过氧化物酶活性增加了140%。同样，心脏缺血区域的超氧化物和组织NO水平分别增加了140%和90%。非缺血区的超氧化物和NO值与假手术对照组相似。总NO合酶（NOS）活性升高了212%，此外，诱导型NOS（iNOS）活性增加6.7倍，在缺血与非缺血区域。MI/R还诱导全身和远端器官（肺）炎症反应。与假手术对照组相比，MI/R大鼠的循环中性粒细胞和血浆NO水平分别增加了163%和138%。肺组织中NO水平和超氧化物生成分别增加了90%和176%。用多克隆抗iNOS和单克隆抗硝基酪氨酸抗体进行免疫组化染色，分别显示iNOS的表达和过氧亚硝基阴离子的产生。MI/R动物的心室壁和肺组织的缺血区域的切片显示出与抗iNOS和抗硝基酪氨酸抗体的显著免疫反应性，表明iNOS和硝基酪氨酸的存在。我们的数据表明，NO，超氧化物和过氧亚硝酸盐的形成升高缺血心脏再灌注后，这表明这些炎症介质可能参与MI/R损伤。
In the present study, the contribution of nitric oxide (NO), superoxide, and peroxynitrite to the inflammatory response induced by myocardial ischemia-reperfusion (MI/R) was investigated. Male Sprague-Dawley rats were anesthetized, and the left main coronary artery was ligated for 20 min and reperfused for 5 h. MI/R induced severe arrhythmias, indicated by a significantly elevated arrhythmia score in the MI/R group compared with that in the sham control group. Creatine kinase activity in the left ventricular free wall of the MI/R group was significantly reduced by 38%. In contrast, myeloperoxidase activity in the left ventricular free wall of the MI/R group was increased by 140%. Similarly, superoxide and tissue NO levels in the ischemic region of the heart were increased by 140 and 90%, respectively. Superoxide and NO values in the nonischemic regions were similar to the sham control group. Total NO synthase (NOS) activity was elevated by 212%; moreover, inducible NOS (iNOS) activity increased 6.7-fold in the ischemic vs. nonischemic regions. MI/R also induced both systemic and remote organ (lung) inflammatory responses. Circulating neutrophils and plasma NO levels were increased by 163 and 138%, respectively, in MI/R rats compared with sham control animals. NO levels and superoxide generation were increased by 90 and 176%, respectively, in the lung tissues. The expression of iNOS and peroxynitrite generation were demonstrated by immunohistochemical staining with polyclonal anti-iNOS and monoclonal anti-nitrotyrosine antibodies, respectively. Sections of both the ischemic area of the ventricular wall and the lung tissue of MI/R animals exhibited a marked immunoreactivity with anti-iNOS and anti-nitrotyrosine antibodies, indicating the presence of iNOS and nitrotyrosine. Our data indicate that NO, superoxide, and peroxynitrite formation are elevated after reperfusion of the ischemic heart, suggesting that these inflammatory mediators may be involved in MI/R injury.