联合转染NOS基因、反义ET核酸对移植静脉血管内皮细胞功能影响的研究

The Effects of Combined Treatment of ET Antisense Oligoneuleotide with eNOS Genetic Transfection on the Endothelial Function of Venous Autografts

目的 :从蛋白分子水平探讨联合转染内皮型一氧化氮合酶 (eNOS)基因、反义内皮素 (ET)核酸对移植静脉血管内皮细胞功能的保护作用。方法 :选择 2 0只大鼠 ,随机等分为实验组和对照组 ,均行自体颈静脉腹主动脉移植手术 ,实验组移植颈静脉在腺病毒介导eNOS基因转染后实施移植手术 ,吻合后移植血管周围应用反义ET核酸凝胶涂布 ;对照组移植静脉无eNOS基因转染 ,仅行凝胶涂布 ,无任何药物。利用放射免疫分析法和Griss方法动态观察血浆ET、NO水平 ,并比较两组之间的差别。结果 :实验组血浆ET水平明显低于对照组 ,而NO则高于对照组。实验组于术后 7天接近术前水平 ,而对照组则于术后 14天接近术前水平。结论 :联合转染eNOS基因和反义ET核酸可有效地保护移植血管内皮细胞 。

Objective To observe the effects of combined tredtment of endothelin (ET) antisense oligoneucleotide with adenovirus-mediated endothelial nitric oxide synthase (eNOS) genetic transfection on the endothelial function of venous autografts and any protection for endothlial function.Methods The external jugule veins were autografted into abdominal aorta arteries in 20 Wistar rats, which were divided evenly into experimental and control group at random. The transplanted veins of experimental group were immersed in the adenovirus-mediated eNOS cDNA solution for 15 minutes before anastomosis and coated with ET antisense olioneucleotide glue gel after anastomosis, while the control ones were only immersed in the adenovirus solution for 15 minutes with only plain gel coating postoperatively. The blood samples were taken at 0, 1, 3, 7, 14, 28 days post-operatively and the levels of ET, NO were determined by radioimmunoassay and Griss method. Results The plasma ET levels were significantly lower in the experimental group than those in the contrel. The condition was just opposite for the NO level. Borh levels returned to normal earlier in experimental group (7 days) than in controls (14 days).Conclusion The autograft venous vascular endothelium can be protected and is function restored by ET antisense oligoneuleotide and eNOS gene transfection.