携带eNOS基因的小口径人工血管在犬的体内研究

目的 观察转染内皮型一氧化氮合酶(eNOS)基因的内皮细胞(EC)种植小口径人工血管在实验犬体内的通畅效果.方法 采用"高压灌注两步法"将转染eNOS基因的EC种植于3 mm人工血管;并植入实验犬股动脉,分1、4、12、24周行数字减影血管造影(DSA)、扫描电镜和透射电镜观察植入后情况.结果 转染eNOS基因的EC种植3 mm人工血管,贴附率在90%以上;10 d左右延伸为一层完整连续的内膜.植入犬体内后1周,单纯人工血管组1/3(3/9)血管阻塞;术后4周,单纯人工血管组全部阻塞(9/9),未转染组近半数阻塞(5/10、4/9);术后12周,单纯人工血管组全部阻塞,未转染组几乎全部阻塞(9/9、9/10);术后24周,未转染组全部阻塞,eNOS转染组8/10通畅.电镜检查表明,eNOS转染组人工血管腔面EC排列紧密,形成一层连续完整的内膜,仅见少许红细胞、血小板及白细胞沉积.结论 转染eNOS基因的EC种植小口径人工血管效果满意,植入实验犬体内后通畅效果良好.

Study on small diameter artificial vascular graft with eNOS gene transfected endothelial cells in canine

Objective To investigate the patency efficacy of small diameter artificial vascular graft with eNOS gene transfected endothelial cells in canine. Methods By using the two steps high pressure injection, eNOS gene transfected endothelial cells were planted on artificial vascular graft with a diameter of 3 mm. The artificial vascular grafts were transplanted into canine femoral artery, and the patency of the artery was observed through digital subtracted angiography (DSA) and electron telescope 1, 4, 12 and 24 weeks after the operation. Results The adherence rate of eNOS gene transfected endothelial cells to artificial vascular grafts was up to 90%. For 10 days, the cells extended and formed a continuous intima. One week after the operation, 3/9 grafts were obstructed in the group of simple artificial vascular grafts; 4 weeks after, all of grafts (9/9) were obstructed in the group of simple artificial vascular grafts and almost half grafts (5/10,4/9) in untransfected groups were obstructed, 12 weeks after, all of the grafts were obstructed in the group of simple artificial vascular grafts and in untransfected groups (9/9,9/10); 24 weeks after, all of the grafts were obstructed in the groups of untransfected artificial vascular grafts, while nearly all of grafts (8/10) were open in eNOS groups. Electron microscope scanning showed that endothelial cells of artificial vascular grafts in eNOS transfected groups arranged closely and formed a continuous intima; only few red blood cells, leucocytes, platelets deposited at the surface of endothelial cells in the artificial vascular grafts. Conclusions The patency efficacy of eNOS gene transfected endothelial cells planting on artificial vascular graft is satisfactory. It could provide an experimental basis for further clinical application of the artificial vascular grafts.