MODULATION OF NO WITH PULMONARY GENE TRANSFER

通过肺部基因转移调节 NO

• 批准号: 6725425
• 负责人: Loretta G Que
• 金额: $ 12.49万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6725425
• 关键词: apoptosis; arginase; biotechnology; cell proliferation; chemokine; enzyme activity; free radical oxygen; gene therapy; histology; hyperoxia; immunocytochemistry; inflammation; laboratory rabbit; lung injury; nitric oxide; nitric oxide synthase; oxidative stress; superoxide dismutase; transfection; transfection /expression vector; western blottings

Acute lung injury in the intensive care unit is treated with administration of high inspired oxygen tensions which can be toxic to the lung. Pulmonary oxygen toxicity is generally attributed to the excessive production of reactive oxygen intermediates (RO1), e.g., superoxide (O2), hydrogen peroxide (H2O2), and hydroxyl radicals. Recently nitric oxide (NO) is being used to treat hyperoxic lung disease, but its effect is variable and its role remains controversial. We hypothesize that selective modulation of NO/RO1 in different cellular compartments regulates tissue responses to hyperoxia. In this proposal, we will employ state of the art adenoviral gene transfer technique in vitro and in vivo. The transgenes we have selected for these studies are neuronal nitric oxide synthase (nNOS), extracellular superoxide dismutase (ECSOD), and arginases I and II (AI and AII). Overexpression of these four genes are expected to affect NO and reactive oxygen intermediates (RO1) production in different cell compartments: nNOS (yields intracellular NO/yields 02-), ECSOD (yields extracellular NO/yields 02-), and AI and AII (yields intracellular NO/yields O2-). We will determine how these transgenes modulate, oxidative/nitrosative stress, and NO/ROI mediated inflammation, cellular proliferation, and apoptosis. The specific aims of this proposal are 1) characterize the transgene expression of nNOS, ECSOD, AI, and AII in vitro 2) characterize intratracheal adenoviral gene transfer in normal rabbit lung and 3) evaluate how adenoviral mediated transgenes nNOS, ECSOD, AI, and AII modulate effect of NO in hyperoxia. These studies should not only better unravedl the basic mechanisms of hyperoxic lung injury, but also provide a rationale basis for therapeutic gene transfer using NO. Ultimately we are hopeful that these results may also extend to other oxidant injuries in the lung.

重症监护病房中的急性肺损伤是通过高启发性氧气张力进行治疗的，可能对肺有毒。 肺氧毒性通常归因于活性氧中间体（RO1）的过度产生，例如，超氧化物（O2），过氧化氢（H2O2）和羟基自由基。 最近，一氧化氮（NO）用于治疗高氧化肺部疾病，但其作用是可变的，其作用仍然存在争议。 我们假设不同细胞室中NO/RO1的选择性调节可以调节组织对高氧的反应。在此提案中，我们将在体外和体内采用最先进的腺病毒基因转移技术。 我们为这些研究选择的转基因是神经元一氧化氮合酶（NNOS），细胞外超氧化物歧化酶（ECSOD）和精氨酸酶I和II（AI和AII）。 预计这四个基因的过表达会影响不同细胞室中的NO和活性氧中间体（RO1）产生：NNOS（nNOS（产量为细胞内NO/产量为02-），ECSOD（屈服于细胞外NO/产量为02-），以及AI和AII（ai and AII）（产生细胞内NO/no/no/no/no//not hart hart fulces O2 o2 o2 o2 o2 o2）。 我们将确定这些转基因如何调节，氧化/亚硝化应激以及无/ROI介导的炎症，细胞增殖和凋亡。 该提案的具体目的是1）表征NNOS，Ecsod，AI和AII的转基因表达在体外2）在正常兔肺中运动内腺病毒基因转移的特征和3）评估腺病毒如何介导的Transgenes NNOS NNOS NNOS NNOS，ECSOD，AI和AII调节效应。这些研究不仅应该更好地脱离过多氧肺损伤的基本机制，而且还应为使用NO提供治疗基因转移的理由基础。最终，我们希望这些结果也可以扩展到肺部其他氧化剂损伤。