尿激酶原基因疗法防治小血管吻合口血栓形成和内膜增生的实验研究

Preventive effection of gene therapy with pro uk gene on the formation of thrombi and intimal hyperplasia of vascular anastomotic site

目的 探讨尿激酶原 (Pro- uk)基因疗法对小血管吻合口血栓形成和内膜增生的影响。 方法 切断大鼠颈总动脉 ,进行原位端端吻合 ,向吻合口血管腔内注入复制缺陷的腺病毒载体 Adv5 -CMV溶液 (对照组 )或复制缺陷的腺病毒重组体 Adv5 - CMV- Pro- uk溶液 (治疗组 ) ,使之在腔内滞留 30min。应用纤维蛋白平板溶圈法检测吻合口组织的纤溶酶活力以观察 Pro- uk基因的表达 ,并进行粘附于吻合口同位素标记的血小板计数、吻合口同位素掺入量和病理形态学检测。 结果 治疗组术后 2、7、14和 90 d均检测到纤溶酶活力 ,术后第 7d活力最高 ,对照组未检测到纤溶酶活力。治疗组粘附于吻合口同位素标记的血小板数目、3 H- Td R掺入量、吻合口血栓出现率和内膜面积均明显少于对照组。 结论 将腺病毒载体介质的 Pro- uk基因转入吻合口血管壁中可减少吻合口血小板沉积和粘附 。

Objective TO study the possibility of preventive effection of gene therapy with pro uk gene on the formation of thrombi and intimal hyperplasia of vascular anastomotic site Methods End to end anastomoses were performed in rat carotid arteries Replication defective recombinant adenovirus ADV 5 CMV solution (control group) or Adv 5 CMV Pro uk solution (treatment group) were injected into anastomotic vascular lumen for 30 minutes By means of pro urokinase activity assay,the number detection of Cr 51 labelled platelts accumulating at anastomotic sites, 3 H TDR incorporation detection of anastomotic sites,pathological changes study,the following results were obtained Results The pro uk activity could be detected on the 2 nd,7 th,14 th,90 th day in the treatment group,but could not be detected in the control group The number of platelets accumulating at the anastomotic sites,thrombosis rate, the average intimal area and 3 H TDR incorporation of anastomotic sites were significantly fewer in the treatment group than that in the control group ( P <0 001) Conclusion When adenovirus vector containing pro uk gene were transferred into anastomotic vascular wall,the foreign gene can produce pro uk at anastomotic sites and effectively prevent the formation of thrombosis and intimal hyperplasia