【摘要】 目的 探讨腺相关病毒载体基因转染的有效途径和方法。方法 我们设计肝动脉、门静脉、双重灌注3种途径和传统、循环、夹闭3种转染方法。通过观察肝脏灌注后颜色、检测肝脏功能和肝细胞转染率,确定灌注途径和转染方法,并探讨其安全性。结果 肝脏灌注后颜色无明显差别,无肿胀或花斑出现,血流开放以后肝脏颜色迅速恢复正常,3种灌注途径ALT比较差异无统计学意义(F=0.343,1.265,0.055,P>0.05);1周后肝脏组织均有免疫荧光染色,并且携带增强型绿色荧光蛋白基因的腺相关病毒载体,转染率比较差异无统计学意义(F=0.080,0.091,0.045,P>0.05)。传统法的转染率略低于循环法,而夹闭法的转染率在各时间段均高于前两种方法,差异有统计学意义(F=3.880,2.976,5.129,P<0.05)。3种方法转染率随时间逐渐升高,6周左右达到高峰,以后缓慢下降。结论 经肝动脉灌注是基因转染的有效途径,夹闭法可以提高目的基因的转染率,两者均无肝脏功能损害,腺相关病毒载体的转染呈缓慢、持续的过程。
【关键词】 腺相关病毒载体; 转染
【Abstract】 Objective To investigate the effective route and proper method in transfecting gene into liver graft mediated by adeno-associated virus vector. Methods Three routes including hepatic artery, portal vein and hepatic artery+portal vein, and 3 methods, i.e. routine, circulation and clamping were employed for infusion. The best infusion route and method of gene transfection into liver graft were determined by observing the color change of liver and detecting liv……
<<<<<全文未完,本文约3030个中文字,未计算英文字母、数字>>>>>