组织型纤溶酶原激活物基因疗法防治血管吻合口血栓栓塞.doc

文档介绍：组织型纤溶酶原激活物基因疗法防治血管吻合口血栓栓塞
作者:张杏泉王少东范清宇殷剑宁裘秀春张殿中
【关键词】组织型纤溶酶原激活物
关键词: 组织型纤溶酶原激活物;基因疗法;血管吻合口;血栓栓塞

摘要:目的探讨防止血管吻合口血栓栓塞的新方法,了解组织型纤溶酶原激活物基因疗法防止吻合口血栓栓塞的效果. 方法①将含有t-PA基因的重组质粒pAdCMVt-PA与缺陷型腺病毒大框架pJM17 共转染病毒包装细胞293细胞,获得有感染能力但复制缺陷的重组腺病毒AdCMVt-PA.②实验动物随机分为对照组和治疗组,均用11--PA溶液,对照组注射AdCMV(不含t-PA的空载体).提取吻合口组织的总RNA行RT-PCR,应用发色底物法检测吻合口组织的纤溶酶活力. 结果治疗组RT-,,3,5,7和14日均可检测到纤溶酶活力,但第3日活力最高,对照组未检测到纤溶酶活力. 结论克隆入腺病毒载体的t-PA基因可在吻合口组织中表达出有生物学活性的t-PA,可能有效地抑制吻合口血栓形成,提高吻合口通畅率.

Keywords:t-PA gene;gene therapy;vascular anastomosis;thrombo-embolism

Abstract:AIM To develop a new way to prevent vascular anastomotic sites from thrombo-embolism and to study the effect of gene therapy with tissue-type plasminogen activator(t-PA)gene on the formation of thrombo-embolism of vas-cular anastomotic 1:binant plas-mid pAdCMVt-PA was cotransfected into293cells with pJM17 ,and infectious but replication-deficient AdCMVt-PA was :Rats were randomly divided into control and treatment -0nylone medical suture was ap-plied to perform rat carotid artery end to end the treatment group,AdCMVt-PA solution was injected into vascular anastomotic site while AdCMV(no containing t-PA DNA)solution was injected into the control ┐SULTS When the isolated RNA was performed with RT-PCR, band appeared in t