尿激酶原基因疗法预防小血管血栓形成的实验研究

Experimental study on the prevention of the formation of thrombos is by Pro-uk gene therapy in small vessels

目的 探讨尿激酶原 (Pro uk)基因疗法对损伤和修复的小动、静脉血栓形成的影响。方法 大鼠颈总动脉和颈静脉剪切伤和挤压伤后间断缝合切口 ,向血管腔内注入含巨细胞病毒(CMV)启动子的腺病毒载体 (Adv5 CMV)溶液 (对照组 )或腺病毒重组体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血管壁组织的纤溶酶活力观察Pro uk基因的表达 ,并进行粘附于血管壁同位素标记的血小板计数和病理观察血管腔内有无血栓形成。结果 治疗组术后 2、7、14、90d颈总动脉和颈静脉均检测到纤溶酶活力 ,治疗组颈动脉壁血小板计数 ( 67± 3)× 10 6 明显少于对照组 ( 130± 9)× 10 6 (P <0 .0 1) ,治疗组颈静脉壁血小板计数 ( 5 9± 2 )× 10 6 ,明显少于对照组 ( 163± 14 )× 10 6 (P <0 .0 1) ,不论是颈总动脉还是颈静脉 ,治疗组血栓出现率均显著低于对照组。结论 将腺病毒载体介导的Pro uk基因转入损伤、修复的小血管壁中能有效地防止血栓形成。

Objective To study the possibility of p re ventive effects of Pro-uk gene therapy on the formation of thrombosis in small injured vessels.Methods Wistar rats' carotid arteries and jugular veins w ere injured and repaired.Replication-defective recombinant adenovirus ADV5-CMV solution (control group) or Adv5-CMV-Pro-uk solution (treatment group) were injected into the injured vascular lumen for 30 min.By means of pro-urokinase a ctivity assay plasmin activity at the vascular walls was measured,Pro-uk gene expression detected,the number of ^(51)Cr labelled platelets accumulating a t vas cular walls counted and the formation of thrombosis in the vessels pathologicall y observed.Results The pro-uk activity could be detected on 2nd,7 th,14th,90th day after operation in the carotid arteries and jugular vei ns of the treatment group,but could not be detected in the control group.The nu mber of platelets accumulating at the jugular walls were significantly fewer in the treatment group[(67±3)×10~6 and (59±2)×10~6 ]than in the contro l group [(130±9) ×10~6 and (163±14)×10~6 ],and the rate of thrombosis formation and occlusion rate of injured vessels were significantly fewer in the treatment group than in the control group.Conclusion When adenovi rus vector containing pro-uk gene was transferred into the injured artery and v ein wall,pro-uk gene could produce pro-uk in the vascular walls and effective ly prevent thrombosis.