白蛋白纳米-超声微泡载组织型纤溶酶原激活物基因靶向体系制备和转染方法  

Preparation of ultrasound microbubble cross-linked to albumin nanoparticles packaged with tPA gene plasmid and method of in vivo transfection

作　　者：季军[1] 姬尚义[1] 何霞[1] 令文萍[1] 

机构地区：[1]广东省深圳市孙逸仙心血管医院,广东深圳518020

出　　处：《中国现代医学杂志》2010年第23期3543-3548,共6页China Journal of Modern Medicine

基　　金：国家863计划资助(No:2007AA021803;2007AA021809;2007AA021904)

摘　　要：目的观察构建的白蛋白纳米组织型纤溶酶原激活物(tPA)基因质粒超声微泡载体的体内转染效果。方法选择兔为实验对象,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基因质粒纳米粒进行交联形成靶向超声微泡转基因载体。免疫组织化学法检测心脏、肝脏、腿部骨骼肌和肋骨组织的靶向转染及tPA的有效表达,同时用ELISA法检测血tPA含量和D-二聚体含量。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、肝脏、骨骼肌和肋骨组织后可见有效tPA表达,同时伴有血tPA和D-二聚体含量增高和tPA功能增强,两者从术前的(0.20±0.05)μg/L和(81.76±9.84)μg/L增高至术后4周的(0.44±0.05)μg/L和(669.28±97.74)μg/L。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,为纳米靶向转基因提供了理论和实验方法。【Objective】 To observe the effect of constructed ultrasound microbubble cross-linked to albumin nanoparticles packaged with tPA gene plasmid on the in vivo transfection.【Methods】 The rabbits were chosen for all experiments.A highly expressive gene plasmid for tPA was constructed and packaged into prepared nanoparticles with bovine serum albumin.The albumin nanoparticles packaged with tPA gene plasmid were cross-linked to ultrasound microbubbles prepared with sucrose and bovine serum albumin to form the vector system for tPA gene transfection.The transfection and effective expression of tPA in heart,liver,skeleton muscle on the leg and the cervical rib were detected with polyclonal antibody to tPA using immunohistochemical method,and tPA level and D-dimer content of blood were tested at the same time.【Results】 The effective expression of tPA vector system for tPA gene transfection could be seen in the tissues mentioned above with the increase in tPA level and D-dimer content of the blood from（0.20±0.05） μg/L and（81.76±9.84） μg/L before the operation to the higher levels of（0.44±0.05） μg/L and（669.28±97.74） μg/L after the transfection respectively.【Conclusions】 The vector system of ultrasound microbubble cross-linked to albumin nanoparticles packaged with tPA gene plasmid has been successfully constructed.This provides a new theory and an experimental method for the nanoparticle targeted gene transfection.

关 键 词：组织型纤溶酶原激活物 白蛋白纳米粒 超声微泡 靶向转染 

分 类 号：R543.1[医药卫生—心血管疾病]