The Antiapoptotic Effects on NO in Pulmonary Endothelium

描述（由申请人提供）：肺内皮是高氧肺损伤早期结构和功能变化的部位。部分还原的氧物质的产生和它们的消除之间的不平衡似乎解释了这种病理的发生和/或维持，并且存在证据表明这种损伤可能因部分还原的氮物质而加剧。然而，近年来，很明显，在许多情况下，一氧化氮（NO）实际上可以限制内皮细胞损伤，并作为一种抗炎细胞保护分子。从最近的报道和初步的数据，我们推测，诱导型一氧化氮合酶衍生的NO可能是通过其潜在的抗凋亡作用，通过翻译后机制，涉及S-亚硝基化肺内皮细胞的保护。因此，本研究申请的具体目的是确定：（1）iNOS衍生的NO在影响iNOS-/-小鼠或在其肺内皮中过表达iNOS的小鼠中的肺内皮细胞结构和高氧中的功能中的作用，所述肺内皮细胞结构和功能通过使用PECAM抗体靶向的含有人iNOS的cDNA的聚乙烯亚胺载体的体细胞基因转移来实现;（2）用共聚焦和多光子激光扫描显微镜观察培养的小鼠肺内皮细胞和离体灌流肺内皮细胞中NO信号的锌释放。

DESCRIPTION (provided by applicant): Pulmonary endothelium is the locus of early structural and functional changes in hyperoxic lung injury. An imbalance between the production of partially reduced oxygen species and their elimination appears to account for the genesis and/or maintenance of such pathology and evidence exists to suggest that such injury may be exacerbated by partially reduced nitrogen species. Nonetheless, in recent years, it is apparent that in many conditions, nitric oxide (NO) may actually limit endothelial cell injury and act as an anti-inflammatory cytoprotective molecule. From recent reports and preliminary data, we hypothesize that iNOS derived NO may be protective to lung endothelium by its potential antiapoptotic effect via posttranslational mechanisms involving S-nitrosylation. Accordingly, the specific aims of this fellowship application are to determine the role of: (1) iNOS-derived NO in affecting pulmonary endothelial cell structure and function in hyperoxia in iNOS-/- mice or in mice that overexpress iNOS in their pulmonary endothelium via somatic gene transfer using a PECAM antibody targeted polyethlyenimine vector containing cDNA to human iNOS; (2) zinc release in NO signaling in the pulmonary endothelium of cultured murine lung endothelial cells and isolated perfused lung using confocal and multiphoton laser scanning microscopy.