Gene therapy with inducible nitric oxide synthase protects against myocardial infarction via a cyclooxygenase-2-dependent mechanism
Gene therapy with inducible nitric oxide synthase protects against myocardial infarction via a cyclooxygenase-2-dependent mechanism
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DOI：
10.1161/01.res.0000065441.72685.29
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发表时间：
2003-04-18
影响因子：
20.1
通讯作者：
Bolli, R
中科院分区：
文献类型：
--
作者：
Li, QH;Guo, Y;Bolli, R
尽管诱导型一氧化氮合酶(INOS)亚型介导了晚期预适应(PC)，但iNOS基因转移是否能复制晚期PC的心脏保护作用尚不清楚，该蛋白在心肌缺血中的作用也存在争议。因此，将人iNOS基因克隆到Rous肉瘤病毒(RSV)启动子的后面，构建了缺失E1、E2a和E3区的腺病毒(Ad)5/iNOS。小鼠心肌内注射Ad5/iNOS可增加局部iNOS蛋白表达和活性，并显著缩小梗塞面积。Ad5/iNOS的心肌保护作用至少与PC缺血组相当。INOS表达增加与环氧合酶-2(COX-2)蛋白表达和前列腺素水平升高有关。COX-2选择性抑制剂NS-398可完全阻断Ad5/iNOS的心肌保护作用，表明COX-2是iNOS依赖的心肌保护的下游效应因子。我们的结论是，iNOS(一种通常被认为是有害的酶)的基因转移提供了强大的心脏保护，其程度与晚期PC相当。这是第一个关于上调iNOS本身就足以缩小梗塞面积的报告。这些结果为抗缺血/再灌注损伤的基因治疗提供了原理证据，该基因治疗可以提高局部心肌一氧化氮合酶水平，而不需要持续静脉输注NO供体，也不会改变全身血流动力学。数据还显示iNOS和COX-2之间存在紧密的偶联，前者的诱导导致后者的二次诱导，后者进而介导iNOS的细胞保护作用。我们认为iNOS和COX-2形成一个应激反应功能模块，减轻缺血/再灌注损伤。
Although the inducible isoform of NO synthase ( iNOS) mediates late preconditioning ( PC), it is unknown whether iNOS gene transfer can replicate the cardioprotective effects of late PC, and the role of this protein in myocardial ischemia is controversial. Thus, the cDNA for human iNOS was cloned behind the Rous sarcoma virus (RSV) promoter to create adenovirus (Ad) 5/iNOS lacking E1, E2a, and E3 regions. Intramyocardial injection of Ad5/iNOS in mice increased local iNOS protein expression and activity and markedly reduced infarct size. The infarct-sparing effects of Ad5/iNOS were at least as powerful as those of ischemic PC. The increased iNOS expression was associated with increased cyclooxygenase-2 (COX-2) protein expression and prostanoid levels. Pretreatment with the COX-2 -selective inhibitor NS-398 completely abrogated the infarct-sparing actions of Ad5/iNOS, demonstrating that COX-2 is an obligatory downstream effector of iNOS-dependent cardioprotection. We conclude that gene transfer of iNOS ( an enzyme commonly thought to be detrimental) affords powerful cardioprotection the magnitude of which is equivalent to that of late PC. This is the first report that upregulation of iNOS, in itself, is sufficient to reduce infarct size. The results provide proof-of-principle for gene therapy against ischemia/reperfusion injury, which increases local myocardial NO synthase levels without the need for continuous intravenous infusion of NO donors and without altering systemic hemodynamics. The data also reveal the existence of a close coupling between iNOS and COX-2, whereby induction of the former enzyme leads to secondary induction of the latter, which in turn mediates the cytoprotective effects of iNOS. We propose that iNOS and COX-2 form a stress-responsive functional module that mitigates ischemia/reperfusion injury.