Barbee R W, Stapleton D D, Perry B D, Ré R N, Murgo J P, Valentino V A, Cook J L
Department of Internal Medicine, Alton Ochsner Medical Institutions, New Orleans, Louisiana 70121.
Biochem Biophys Res Commun. 1993 Jan 15;190(1):70-8. doi: 10.1006/bbrc.1993.1012.
We determined the time course of gene expression following DNA/Lipofectin transfection of normal or previously injured arterial segments using direct intraluminal infusion following surgical exposure. Constructs possessing the firefly luciferase cDNA regulated by Simian virus 40, Rous sarcoma virus, or alpha-actin promoter were incubated together with Lipofectin for 30 minutes. Arterial segments were assayed for luciferase activity following harvest at 2-21 days. Without prior injury, luciferase activity was only 2.5-fold greater than background two days following gene transfer. Arterial injury three days before gene transfer resulted in luciferase activity 12.5-fold over background levels. This observation has clinical implications with regard to gene therapy following angioplasty, a procedure that is associated with endothelial cell denudation and smooth muscle cell proliferation. Maintenance of gene expression for several days could ameliorate the early smooth muscle migration and proliferation following arterial injury.
我们通过手术暴露后直接腔内灌注，确定了正常或先前损伤的动脉节段经DNA/脂质体转染后基因表达的时间进程。将含有由猿猴病毒40、劳氏肉瘤病毒或α-肌动蛋白启动子调控的萤火虫荧光素酶cDNA的构建体与脂质体一起孵育30分钟。在2至21天收获后，对动脉节段进行荧光素酶活性测定。在没有先前损伤的情况下，基因转移两天后荧光素酶活性仅比背景高2.5倍。基因转移前三天的动脉损伤导致荧光素酶活性比背景水平高12.5倍。这一观察结果对于血管成形术后的基因治疗具有临床意义，血管成形术与内皮细胞剥脱和平滑肌细胞增殖有关。基因表达维持数天可改善动脉损伤后早期平滑肌的迁移和增殖。
