Adenoviral-mediated transfer of the human endothelial nitric oxide synthase gene reduces acute hypoxic pulmonary vasoconstriction in rats
Adenoviral-mediated transfer of the human endothelial nitric oxide synthase gene reduces acute hypoxic pulmonary vasoconstriction in rats
复制标题
DOI：
10.1172/jci118795
复制
发表时间：
1996-07-15
影响因子：
15.9
通讯作者：
Collen, D
中科院分区：
文献类型：
--
作者：
Janssens, SP;Bloch, KD;Collen, D
一氧化氮(NO)是一种参与肺血管张力调节的血管扩张剂，由一氧化氮合酶(NOS)家族合成。为了研究腺病毒介导的内皮型一氧化氮合酶(CeNOS)过表达是否能减轻缺氧性肺血管收缩反应，我们将含有ceNOS基因的重组腺病毒(AdCMVceNOS)的3×10(9)个空斑形成单位气雾化注入大鼠肺内。感染4天后，用免疫印迹技术证实转基因表达。在AdCMVceNOS转导肺的肺泡和中、小肺血管内可见弥漫性的ceNOS免疫染色。与对照组相比，AdCMVceNOS组大鼠肺组织[H-3]精氨酸到[H-3]瓜氨酸的转化率增加了86%，肺组织cGMP水平从7+/-1增加到59+/-9pmol/mg蛋白(P<；0.05)。在急性低氧(FiO2=0.10)25min时，雾化吸入生理盐水的大鼠平均肺动脉压(PAP)从17+/-1升至27+/-1 mm Hg(n=4)，而携带核靶向β-半乳糖苷酶基因的腺病毒载体(AdCMV beta Gal，n=8)的大鼠平均肺动脉压(PAP)从18+1升至28+/-1 mm Hg。相反，在AdCMVceNOS转导的大鼠(n=8)，低氧引起的PAP升高显著减弱(18+/-1至23+/-2毫米汞柱)。全身血压不受气雾剂基因转移的影响。因此，腺病毒介导的ceNOS基因转移到大鼠肺中增加了ceNOS的表达和活性，并减少了急性缺氧性肺血管收缩。雾化高表达血管扩张蛋白的重组腺病毒可以作为一种选择性的肺血管扩张剂，有望成为未来治疗肺动脉高压的策略。
Nitric oxide (NO), a vasodilator involved in the regulation of pulmonary vascular tone, is synthesized by a family of enzymes, nitric oxide synthases (NOS). To investigate whether adenoviral-mediated overexpression of constitutive endothelial NOS (ceNOS) would attenuate hypoxic pulmonary vasoconstriction, we aerosolized 3 x 10(9) plaque forming units of a recombinant adenovirus containing the ceNOS gene (AdCMVceNOS) into rat lungs. Four days after infection, transgene expression was confirmed using immunoblot techniques. Diffuse ceNOS immunostaining was detected in alveoli and medium-sized and small pulmonary vessels of AdCMVceNOS-transduced lungs. AdCMVceNOS-transduction was associated with an 86% increase in [H-3]arginine to [H-3]citrulline conversion and a rise in pulmonary cGMP levels from 7+/-1 to 59+/-9 pmol/mg protein in lungs from AdCMVceNOS versus control rats, (P < 0.05). During acute hypoxia (FIO2 = 0.10) for 25 min, mean pulmonary artery pressure (PAP) increased significantly from 17+/-1 to 27+/-1 mmHg in rats aerosolized with saline (n = 4) and from 18+1 to 28+/-1 mmHg in rats given an adenoviral vector expressing a nuclear-targeted beta-galactosidase gene (AdCMV beta gal, n = 8). In contrast, in AdCMVceNOS-transduced rats (n = 8) the hypoxia-induced increase in PAP was significantly attenuated (18+/-1 to 23+/-2 mmHg). Systemic blood pressure was not affected by aerosol gene transfer. Thus, adenoviral-mediated ceNOS gene transfer to rat lungs increases ceNOS expression and activity, and reduces acute hypoxic pulmonary vasoconstriction. Aerosolized recombinant adenovirus overexpressing vasodilatory proteins can act as a selective pulmonary vasodilator and may hold promise as a future therapeutic strategy for pulmonary hypertension.