The Antiapoptotic Effects on NO in Pulmonary Endothelium

NO对肺内皮细胞的抗凋亡作用

• 批准号: 6694444
• 负责人: ANNETTE S WILSON
• 金额: $ 5万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6694444
• 关键词: apoptosis; biological signal transduction; confocal scanning microscopy; cytoprotection; fluorescence resonance energy transfer; genetically modified animals; immunocytochemistry; laboratory mouse; nitric oxide; nitric oxide synthase; northern blottings; postdoctoral investigator; respiratory epithelium; tissue /cell culture; western blottings; zinc

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.

描述（申请人提供）：肺内皮是高氧肺损伤早期结构和功能改变的位点。部分还原的氧的产生和消除之间的不平衡似乎解释了这种病理的发生和/或维持，有证据表明，部分还原的氮可能加剧这种伤害。尽管如此，近年来，很明显，在许多情况下，一氧化氮（NO）实际上可能限制内皮细胞损伤，并作为抗炎细胞保护分子。根据最近的报道和初步数据，我们假设iNOS衍生的NO可能通过涉及s -亚硝基化的翻译后机制具有潜在的抗凋亡作用，从而对肺内皮具有保护作用。因此，本研究申请的具体目的是确定iNOS来源的NO在高氧条件下对iNOS-/-小鼠或在肺内皮中过度表达iNOS的小鼠的肺内皮细胞结构和功能的影响，通过使用PECAM抗体靶向含有cDNA的聚亚胺载体向人iNOS进行体细胞基因转移；(2)利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注肺内皮细胞NO信号中锌的释放。