Investigation of Regression of Atherosclerotic Lesion through Regulation of Nitric Oxide Related Responses -Treatment by Gene Transfer of Nitric Oxide Synthase-

目的：探讨内皮型和诱导型一氧化氮合酶(eNOS，iNOS)基因转染腺病毒载体对动脉粥样硬化晚期病变的影响。我们制备了内皮型一氧化氮合酶和诱导型一氧化氮合酶的腺病毒载体。用球囊导管损伤40只雄性和去卵巢雌性兔的腹主动脉，给予HCD[标准饲料+1%胆固醇]喂养12周。治疗结束后，用血管内超声(IVUS)确定胸主动脉(HCD诱导)和腹主动脉(内皮损伤+HCD)动脉粥样硬化的大小和性质。将eNOS、iNOS及eNOS+iNOS基因转移到胸、腹主动脉粥样硬化病变处，采用DISTH血流预留导管。以β-半乳糖苷酶基因表达载体为对照。在基因转导3天和7天后，用血管内超声检查胸、腹主动脉粥样硬化的大小和性质的变化，然后处死动物，取…更多和考查如下。通过NO选择性电极以及血管反应和血浆NO代谢产物来评估基础和刺激后的NO释放。此外，还评估了动脉粥样硬化病变的表面受累程度和面积。进行免疫组织化学研究。对巨噬细胞源性泡沫细胞占据的面积进行编号。HCD介导的血脂水平升高不受各载体基因转移的影响。内皮型一氧化氮合酶和诱导型一氧化氮合酶基因转移不仅可部分改善胸主动脉粥样硬化，还可部分改善腹主动脉重度动脉硬化。HCD致兔球囊损伤后，乙酰胆碱诱导的NO介导的松弛作用受到严重损害。内皮型一氧化氮合酶和诱导型一氧化氮合酶基因转移可轻微恢复球囊损伤和添加致动脉粥样硬化饲料的兔腹主动脉松弛功能。以上结果提示，eNOS和iNOS基因转移可以逆转严重机制的晚期病变，更有效的联合病毒载体有待于进一步阐明。较少

We determined the effect of endothelial and inducible nitric oxide synthase (eNOS, iNOS) gene transfer using adenovirus vector on the advanced lesion of atherosclerosis. We prepared adenovirus vector of eNOS and iNOS cDNA. The abdominal aortae of 40 male and oophorectomized female rabbits were injured by balloon catheter and they were fed with HCD [standard diet and 1% cholesterol] for 12 weeks. After treatment, we made sure the size and character of atherosclerosis in thoracic (HCD induced) and abdominal (endothelial injury plus HCD) aorta using IVUS (intra vascular ultra sonography). Then, we transferred gene of eNOS, iNOS and eNOS plus iNOS onto atherosclerotic lesion of thoracic and abdominal aorta using dispatch blood flow reserved catheter. As the control, cDNA vector of β-galactosidase was transferred. After 3 and 7 days of gene transfer, the change of size and character of atherosclerosis in thoracic and abdominal aorta was examined using IVUS, and then, animals were sacrificed … More and examined as below. Basal and stimulated NO release was estimated by an NO selective electrode as well as vascular response and the plasma NO metabolites. Surface involvement and area occupied by atherosclerotic lesion were also evaluated. An immunohistochemical study was done. The area occupied by macrophage derived foam cells was numbered. HCD mediated increased plasma lipid levels were not affected by gene transfer of each vector. Not only atherosclerosis in the thoracic aorta but also severe atherosclerosis in the abdominal aorta was partially improved by eNOS and iNOS gene transfer. The acetylcholine-induced NO-mediated relaxation was severely impaired in HCD supplemented balloon injured rabbits. The impaired abdominal aortic relaxation of balloon injured and atherogenic diet supplemented rabbits was slightly restored by eNOS and iNOS gene transfer. The above results suggest that gene transfer of eNOS and iNOS could regress the advanced lesion of severe mechanisms and more effective combined virus vector will be elucidated from now. Less