组织型纤溶酶原激活剂基因逆转录病毒载体兔左心耳内靶向溶栓研究  

Study on target thrombolytic effect for the tissue-type plasminogen activator (tPA) gene retroviral vector in the left atrium chip implanted the Dacron thrombosis model in rabbits

作　　者：罗洪波[1] 向道康[1] 蔡鸿[2] 冯超[1] 向龙 

机构地区：[1]贵州省人民医院心外科,贵州贵阳550002 [2]贵州省人民医院神经内科,贵州贵阳550002

出　　处：《贵州医药》2017年第11期1132-1134,F0003,共4页Guizhou Medical Journal

基　　金：贵州省科技计划项目[黔科合LH号(2016)7181]

摘　　要：目的观察组织型纤溶酶原激活剂(tPA)基因局部转导对特氟隆(Dacron)片(与机械瓣瓣环同质)的溶血栓作用。方法 30只兔建立左心耳内Dacron片植入血栓模型,随机分为PLEGFP-N1-tPA治疗组(n=10)、PLEGFP-N1空载体对照组(n=10)、空白对照组(n=10),局部基因转导,于术后2,30d各组1/2动物取材(6个亚组A1、B1、C1、A2、B2、C2),聚光共聚焦(confocal)观察所取左心耳组织增强型绿色荧光蛋白(EGFP)表达,体视镜和电视镜观察Dacron片表面血栓情况,免疫印迹(Western blot)、血浆平板和酶联免疫吸附实验(ELISA)分别检测所取组织tPA表达、溶栓情况。结果术后2,30d取材,治疗组所取左心耳组织confocal均观察到多而强的EGFP表达,Plegfp-N1对照组也有EGFP表达,但空白对照组无EGFP表达。30个Dacron片加未植入组5个Dacron片共35个,在体视镜(×160)和电视镜(×500)下观察,未植入组和治疗组Dacron片表面均无血栓,对照组表面均有血栓。治疗组所取静脉Western blot检测均有外源tPA表达,对照组未检测到。血浆平板显示:治疗组均有大的溶解圈,有明显溶栓作用,对照组没有。结论 Plegfp-N1-tPA局部基因转导能有效阻止外源移植物表面血栓形成。Objective To investigate the tissue-type plasminogen activator （tPA） gene transduction on local Tel ion （Dacron） film （with mechanical prosthetic ring homogeneity） of the thrombolytic effect. Methods 30 rabbits to establish chip implanted in the left atrium Dacron thrombosis were randomly divided into PLEGFP-NI-tPA treatment group （n=10）,PLEGFP N1 empty vector control group （n=10）,the control group （n=10）,local gene transduction,in half of each group after 2,30 d animal derived （6 subgroups A1,B1,CI,A2,B2,C2）,condenser confocal observation of the left atrial tissue obtained enhanced green fluorescent protein （EGFP） expression,stereoscope mi croscope Dacron film and television thrombus surface conditions,immunoblotting （Western blot）, plasma flat-panel and ELISA （EI.ISA） of tissue were used to detect the expression of tPA,dissolved bolt. Results After 2 d and 30 d, taken from the treatment group were observed in left atrial tissue confocal to the many and strong EGFP expression, Plegfp-Nl control group also expressed EGFP,but no EGFP expression in the control group. Plus 30 Dacron film 5 is not implanted Dacron sheet of group 35,in the stereoscope （X160 times） and TV lens （X500 times） were observed, no implant group and the no treatment group Dacron film surface thrombus,were Groups are thrombus surface. Vein taken from the treatment group were detected by Western blot of exogenous tPA expression in the control group was not detected. Plasma flat panel display： the dissolution of the treatment group had a large circle, with a clear thrombolysis,there is no control group. Conclusion Plegfp-Nl-tPA local gene transfer can effectively prevent the surface of the foreign graft thrombosis.

关 键 词：逆转录病毒科 纤溶酶原 基因治疗 血栓 

分 类 号：R541.4[医药卫生—心血管疾病]