INOS GENE TRANSFECTION IN PULMONARY HYPERTENSION

肺动脉高压（PH）是发病的重要原因， 影响广泛的患者的死亡率。 新生儿肺 高血压是新生儿入院的第二大原因， 重症监护室进行呼吸支持 在成年人中，PH导致 慢性阻塞性肺疾病患者的发病率和死亡率 肺部疾病 在所有患者中，PH的特征是细胞 肺血管床的增生和血管反应性改变。 本提案的目的是评价血管扩张剂的疗效 病毒介导的诱导型一氧化氮产生的NO和毒性 在肺中的iNOS基因转染，并确定 病毒转染的iNOS对PH发病机制的影响。 一般的假设是病毒转染的iNOS将导致 足够的NO形成以调节肺血管收缩， 减轻与肺动脉高压相关的肺血管变化 高血压，但NO形成不足导致毒性。 利用人iNOS基因，以E. colilac Z报告基因 我们构建了编码β-半乳糖苷酶（β-gal）的基因腺病毒， 本研究提出的目标是：1）优化iNOS基因的传递 和在大鼠肺中的表达，2）确定转染的 iNOS对肺动脉高压形成的影响; 3）比较 血管内和血管内递送iNOS基因， 基因表达、血管反应性和毒性。 这些目标是 具体目标1：评估 腺病毒介导的iNOS基因转染对人肝癌细胞增殖的影响 减弱急性肺血管收缩反应。 具体目标 编号2：评估iNOS基因转染介导的对 慢性缺氧诱导的肺动脉高压的发展。 具体 目标3：评估血管内和静脉内给药的疗效和毒性。 气管内施用腺病毒iNOS构建体。 该方法将涉及使用含有该基因的腺病毒构建体 用于血管内给药的iNOS或β-gal; 然后研究肺以确定血管反应性，NO 转染的iNOS的产生和定位。 有些老鼠会 转染并暴露于慢性缺氧。 最后，血管内和 将在基因定位方面比较肠内递送 和毒性。

Pulmonary hypertension (PH) is a significant cause of morbidity and mortality affecting a broad range of patients. Neonatal pulmonary hypertension is the second leading cause for admission to neonatal intensive care units for respiratory support. In adults, PH causes significant morbidity and mortality in patients with chronic obstructive pulmonary disease. In all patients, PH is characterized by cellular proliferation and altered vasoreactivity in the pulmonary vascular bed. The objectives of this proposal are to evaluate the vasodilator efficacy and toxicity of NO produced by virally mediated inducible nitric oxide synthase (iNOS) gene transfection in the lung and to determine the effect of virally transfected iNOS on the pathogenesis of PH. The general hypothesis is that virally transfected iNOS will result in sufficient NO formation to modulate pulmonary vasoconstriction and attenuate pulmonary vascular changes associated with pulmonary hypertension, but insufficient NO formation to result in toxicity. Utilizing human iNOS gene and, as a control, the E. coli lac Z reporter gene coding for beta-galactosidase (beta-gal) adenovirus constructs our goals set forth in this proposal are: 1) to optimize iNOS gene delivery and expression in the rat lung, 2) to determine the role of transfected iNOS on the development of pulmonary hypertension, and 3) to compare intravascular and intratracheal delivery of the iNOS gene in terms of gene expression, vascular reactivity and toxicity. These goals are addressed in the following specific aims: Specific Aim number 1: Assess the effectiveness of adenovirus-mediated iNOS gene transfection in attenuating acute pulmonary vasoconstrictor responses. Specific Aim number 2: Assess iNOS gene transfection-mediated effects on the development of chronic hypoxia-induced pulmonary hypertension. Specific Aim number 3: Assess the efficacy and toxicity of intravascularly and intratracheally administered adenoviral iNOS constructs. The methods will involve using adenovirus constructs containing the gene for iNOS or beta-gal that will be administered intravascularly; the lungs will then be studied to determine vascular reactivity, NO production, and localization of transfected iNOS. Some rats will be transfected and exposed to chronic hypoxia. Finally, intravascular and intratracheal delivery will be compared in terms of gene localization and toxicity.