白蛋白纳米-超声微泡载组织型纤溶酶原激活物基因预防心脏瓣膜置换术血栓形成的实验研究  

Experimental study on an ultrasound microbubble crosslinked to albium nanoparticle packaged with tPA gene plasmid to prevent the thrombosis after heart valve replacement

作　　者：季军[1] 姬尚义[1] 何霞[1] 令文萍[1] 潘一峰[2] 

机构地区：[1]深圳市孙逸仙心血管医院,广东深圳518020 [2]中南大学卫生部肝胆肠外科研究中心,湖南长沙410008

出　　处：《中国现代医学杂志》2011年第26期3213-3218,3223,共7页China Journal of Modern Medicine

基　　金：国家863项目(No:2007AA021803;2007AA021809;2007AA021904)

摘　　要：目的观察构建的白蛋白纳米-超声微泡载组织型纤溶酶原激活物(tPA)基因预防心脏瓣膜置换术血栓形成的效果。方法以狗作为实验对象;构建高表达tPA基因质粒;以白蛋白为原料制备载tPA基因质粒纳米粒;用蔗糖和白蛋白制备糖蛋白超声微泡并与载tPA基因质粒纳米粒交联形成靶向超声微泡转基因载体。在给狗实施三尖瓣机械瓣膜置换术后行心脏超声靶向转基因并观察对术后血栓形成的预防作用。用免疫组织化学法检测心脏组织tPA抗原表达并用ELISA法检测血tPA和D-二聚体含量的变化。结果构建的白蛋白纳米tPA基因质粒超声微泡载体系统超声靶向转染心脏后4周,靶向心肌组织表达tPA抗原,同时伴有血tPA和D-二聚体含量增高及tPA功能增强,有效预防了心脏三尖瓣机械瓣膜置换术血栓的形成。结论成功建立了白蛋白纳米tPA基因质粒超声微泡载体系统,为血栓相关性疾病的预防提供了实验依据。【Objective】 To observe the effect of constructed ultrasound microbubble crosslinked to albium nanoparticle packaged with tPA gene plasmid on prevention of the thrombosis after heart valve replacement.【Methods】 A dog model of mechanical tricuspid valve replacement was constructed.A highly expressive gene plasmid for tPA was constructed and packaged into prepared namoparticle with bovine serum albumin.This albium nanoparticle packaged with tPA gene plasmid was crosslinked to an ultrasound microbubble prepared with sucrose and bovine serum albumin to form the targeting vector system for tPA gene transfection to the myocardium with a ultrasound therapy after the tricuspid valve replacement.The transfection and effective expression of tPA in myocardium were detected with polyclonal antibody to tPA using indirect immunohistochemical method and tPA level and D-dimer content of blood were tested with the method of enzyme linked immunosorbent assay before and 4 week after operation.【Results】 The effective expression of tPA vector system for tPA gene transfection could seen in myocardium with the increase in tPA level and D-dimer content of the blood and the thrombosis was prevented 4 week after operation.【Conclusions】 The vector system of ultrasound microbubble crosslinked to albium nanoparticle packaged with tPA gene plasmid was contructed successfully.This provides a experimental basis for the prevention of throbosis-related diseases.

关 键 词：组织型纤溶酶原激活物 白蛋白纳米粒 白蛋白超声微泡 瓣膜置换术 血栓形成 

分 类 号：R-332[医药卫生] R619.2