Channon K M, Qian H, Neplioueva V, Blazing M A, Olmez E, Shetty G A, Youngblood S A, Pawloski J, McMahon T, Stamler J S, George S E
Divisions of Cardiology, and Pulmonology, Departments of Medicine and Pharmacology, Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710, USA.
Circulation. 1998 Nov 3;98(18):1905-11. doi: 10.1161/01.cir.98.18.1905.
The vascular endothelium is anatomically intact but functionally abnormal in preatherosclerotic states, and an early deficit in the bioavailability of nitric oxide (NO) or related molecules has been described in both humans and animal models. We hypothesized that the targeted gene transfer of NO synthase (NOS) isoforms might ameliorate or reverse the deficit.
We constructed a recombinant adenovirus, Ad.nNOS, that expresses the neuronal isoform of NOS (nNOS) and used it for in vivo endovascular gene transfer to carotid arteries (CA) from normal and cholesterol-fed rabbits. Vessels were harvested 3 days after gene transfer. In CA from normal rabbits, Ad.nNOS generated high levels of functional nNOS protein predominantly in endothelial cells and increased vascular NOS activity by 3.4-fold relative to sham-infected control CA. Ad.nNOS gene transfer also significantly enhanced endothelium-dependent vascular relaxation to acetylcholine; at 3 micromol/L acetylcholine, Ad.nNOS-treated arteries showed an 86+/-4% reduction in precontracted tension, whereas control CA showed a 47+/-6% reduction in tension. Contraction in response to phenylephrine and relaxation in response to nitroprusside were unaffected in both control and Ad.nNOS-treated CA. To determine the effect of Ad.nNOS in atherosclerotic arteries, 10 male New Zealand White rabbits maintained on a 1% cholesterol diet for 10 to 12 weeks underwent gene transfer according to the same protocol used in normal rabbits. Ad.nNOS-treated arteries showed a 2-fold increase in NADPH-diaphorase staining intensity relative to sham-infected and Ad. betaGal-treated arteries. The CA from cholesterol-fed rabbits showed impaired acetylcholine-induced relaxation, but this abnormality was almost entirely corrected by Ad.nNOS gene transfer.
In vivo adenovirus-mediated endovascular delivery of nNOS markedly enhances vascular NOS activity and can favorably influence endothelial physiology in the intact and atherosclerotic vessel wall.
在动脉粥样硬化前期状态下，血管内皮在解剖结构上保持完整，但功能异常，并且在人类和动物模型中均已发现一氧化氮（NO）或相关分子的生物利用度早期存在缺陷。我们推测，一氧化氮合酶（NOS）亚型的靶向基因转移可能会改善或逆转这种缺陷。
我们构建了一种表达神经元型NOS（nNOS）的重组腺病毒Ad.nNOS，并将其用于正常和喂食胆固醇的兔子颈动脉的体内血管内基因转移。基因转移后第3天收获血管。在正常兔子的颈动脉中，Ad.nNOS主要在内皮细胞中产生高水平的功能性nNOS蛋白，并且相对于假感染对照颈动脉，血管NOS活性增加了3.4倍。Ad.nNOS基因转移还显著增强了对乙酰胆碱的内皮依赖性血管舒张；在3 μmol/L乙酰胆碱作用下，经Ad.nNOS处理的动脉在预收缩张力下降低了86±4%，而对照颈动脉降低了47±6%。对照和经Ad.nNOS处理的颈动脉对去氧肾上腺素的收缩反应和对硝普钠的舒张反应均未受影响。为了确定Ad.nNOS在动脉粥样硬化动脉中的作用，10只雄性新西兰白兔在1%胆固醇饮食下饲养10至12周，按照与正常兔子相同的方案进行基因转移。相对于假感染和Ad.βGal处理的动脉，经Ad.nNOS处理的动脉NADPH-黄递酶染色强度增加了2倍。喂食胆固醇的兔子的颈动脉显示乙酰胆碱诱导的舒张受损，但这种异常几乎完全通过Ad.nNOS基因转移得到纠正。
体内腺病毒介导的nNOS血管内递送显著增强血管NOS活性，并可对完整和动脉粥样硬化血管壁中的内皮生理产生有利影响。
