摘要
目的:研究半乳糖基化壳聚糖(Galactosylated chitosan, GC)在狗体内的肝靶向性作用. 方法:GC与质粒pEGFP-N1混和制备成纳米微囊复合物,体外转染SMMC-7721细胞.杂种狗术前1 d静脉注射氯化钆(14 mg/kg)以清除Kupffer细胞.将含1 mg质粒的纳米微囊经肝动脉和门静脉注射入动物体内.48 h 后取主要脏器组织做冰冻切片,在荧光显微镜下观察绿荧光蛋白表达情况.以裸质粒作为对照. 结果:7721细胞中有绿荧光蛋白的表达,体内实验GC组肝组织中有绿荧光蛋白表达,其他脏器组织中仅有微量表达. 结论:半乳糖基化壳聚糖在体外有较高的转染率,在大动物体内有肝靶向性作用,肝动脉、门静脉联合给药是基因治疗的一种有效途径.
AIM: To investigate the hepatocyte-targeting effect of galactosylated chitosan (GC) in vivo. METHODS: GC was complexed with plasmid pEGFP-N1 to develop nanoparticles, which were then transfected into SMMC-7721 cells. Dogs were treated with gadolinium chloride (14 mg/kg) via intravenous injection 24 hours before operation to deplete Kupffer cells. The nanoparticles containing 1 mg plasmid were injected through hepatic artery and portal vein. Main organs were collected and used to prepare frozen sections. Green fluorescence was examined under the fluorescence microscope after 48 hours. Naked plasmid DNA was used as control. RESULTS: Green fluorescent protein was detected in SMMC-7721 cells transfected with pEGFP-N1 plasmid.The transfection efficiency was higher in GC group (60.4±2.2%), than the liposome group (55.6±1.6%) (P= 0.04). In vivo experiments showed that in the GC group, bright green fluorescence existed in the hepatic tissue, while only weak fluorescence was observed in other tissues. CONCLUSION: GC nanoparticle can be efficiently trans-fected in vitro. It is hepatocyte-specific in large animals. The administration via hepatic artery and portal vein is an effective approach for gene transfer.
出处
《世界华人消化杂志》
CAS
北大核心
2005年第7期848-851,共4页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.30070210~~
