内皮型一氧化氮合酶基因转染在兔颈静脉移植血管桥中的表达

Expression of transfected endothelial nitric oxide synthase transfer on carotid vein graft in rabbits

目的:研究腺病毒载体介导的人内皮型一氧化氮合成酶基因在兔自体移植颈静脉中的表达,寻找预防移植血管再狭窄的方法.方法:建立兔颈外静脉颈总动脉旁路移植模型,18只兔随机等分为3组,A:对照组,B:空载腺病毒液感染组,c:eN0s基因转染组.在兔颈外静脉颈总动脉旁路移植术中,分别应用空载腺病毒液或AdcMveN0s血管桥内注入法感染移植静脉1 h.术后4wk,应用多聚寡核普酸探针原位检测方法检测外源性基因mRNA的表达;免疫组织化学染色方法检测eN0s蛋白的定位;HE及弹力纤维染色后,应用计算机图像分析系统检测移植静脉内膜、中膜增生情况.结果:人内皮型一氧化氮合成酶基因在自体移植静脉中表达出相应的mRNA和蛋白质;术后4 wk,与对照组相比,空载腺病毒液感染组移植静脉内膜和中膜厚度无明显变化;eN0s基因转染组移植静脉内膜和中膜厚度分别减少42.3%,13.5%,内膜厚度冲膜厚度比值(l/M)减少34.6%.结论:腺病毒载体介导的人内皮型一氧化氮合成酶基因成功转染兔自体移植颈静脉中并有效表达,eN0s基因具有防治移植静脉内膜增生作用.

AIM: To study the expression of adenoviral-mediated endothelial nitric oxide synthase gene (AdCMVeNOS) on carotid vein graft in rabbits and to find the way to prevent vein graft restenosis. METHODS: External jugular vein-common carotid artery bypass graft model was established. Eighteen rabbits were randomly divided into 3 groups: Control group (A), simple adenovirus infection group (B) and eNOS gene transfection group (C). After four weeks, the expression of exogenous eNOS mRNA and protein was detected by in situ hybridization and immunohistochemistry respectively. The proliferation of intima and media in vein graft was measured by computer graph-analyzing system. RESULTS: The expressions of exogenous eNOS mRNA and protein were detected in vein graft. There was no obvious difference in the thickness of intima and media between control group and simple adenoviral infection group. Compared with that in control group, the thickness of intima and media in eNOS gene transfection group decreased by 42.3% and 13.5% respectively. The ratio of the thickness of intima to media (I/M ) decreased by 34.6%. CONCLUSION: The transfected eNOS gene can be expressed on carotid vein graft in rabbits. eNOS gene has some protective effect on vein graft restenosis.