Inactivity of nitric oxide synthase gene in the atherosclerotic human carotid artery
Inactivity of nitric oxide synthase gene in the atherosclerotic human carotid artery
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DOI：
10.1007/s00395-007-0650-7
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发表时间：
2007-07-01
影响因子：
9.5
通讯作者：
Luescher, Thomas F.
中科院分区：
文献类型：
--
作者：
Tanner, Felix C.;van der Loo, Bernd;Luescher, Thomas F.
目的 一氧化氮 (NO) 抑制血栓形成、血管收缩和平滑肌细胞增殖。我们研究了动脉粥样硬化人颈动脉离体内皮NO合酶（eNOS）基因转移后NO释放是否增强。方法和结果 Western blotting 和免疫组织化学显示转导增强了 eNOS 表达；然而，卟啉微传感器测量的亚硝酸盐产生和一氧化氮释放都没有改变。相反，转导增强了非动脉粥样硬化大鼠主动脉和人乳内动脉中 NO 的产生。在转导的颈动脉中，钙依赖性 eNOS 活性极小，并且与对照条件没有差异。实验组之间的血管四氢生物蝶呤浓度没有差异。用 FAD、FMN、NADPH、L-精氨酸以及墨蝶呤或四氢生物蝶呤处理转导的颈动脉不会改变 NO 释放。转导颈动脉和对照中的超氧化物形成相似。用超氧化物歧化酶(SOD)、PEG-SOD、PEG-过氧化氢酶处理转导的颈动脉不影响NO释放。结论 eNOS 在动脉粥样硬化人颈动脉中转导导致高表达，但重组蛋白没有任何可测量的活性。动脉粥样硬化血管的缺陷既不是由辅因子缺乏引起的，也不是由一氧化氮分解增强引起的。由于血管成形术是在动脉粥样硬化的动脉中进行的，因此 eNOS 基因治疗不太可能提供临床益处。
Objective Nitric oxide (NO) inhibits thrombus formation, vascular contraction, and smooth muscle cell proliferation. We investigated whether NO release is enhanced after endothelial NO synthase (eNOS) gene transfer in atherosclerotic human carotid artery ex vivo. Methods and Results Western blotting and immunohistochemistry revealed that transduction enhanced eNOS expression; however, neither nitrite production nor NO release measured by porphyrinic microsensor was altered. In contrast, transduction enhanced NO production in non-atherosclerotic rat aorta and human internal mammary artery. In transduced carotid artery, calcium-dependent eNOS activity was minimal and did not differ from control conditions. Vascular tetrahydrobiopterin concentrations did not differ between the experimental groups. Treatment of transduced carotid artery with FAD, FMN, NADPH, L-arginine, and either sepiapterin or tetrahydrobiopterin did not alter NO release. Superoxide formation was similar in transduced carotid artery and control. Treatment of transduced carotid artery with superoxide dismutase (SOD), PEG-SOD, PEG-catalase did not affect NO release. Conclusions eNOS transduction in atherosclerotic human carotid artery results in high expression without any measurable activity of the recombinant protein. The defect in the atherosclerotic vessels is neither caused by cofactor deficiency nor enhanced NO breakdown. Since angioplasty is performed in atherosclerotic arteries, eNOS gene therapy is unlikely to provide clinical benefit.