靶向性脂质体P-MMA-DOSPER介导pEGFP-C2脑内转染  

Transfection of pEGFP-C2 in Brain Mediated by Targeting Liposome P-MMA-DOSPER

作　　者：唐劭年[1] 刘振华[1] 赵连旭[1] 邹志浩[2] 杜谋选[2] 

机构地区：[1]南方医科大学附属珠江医院神经内科,广州510282 [2]南方医科大学附属珠江医院神经医学研究所,广州510282

出　　处：《生物医学工程学杂志》2008年第5期1170-1174,共5页Journal of Biomedical Engineering

摘　　要：本研究以衍生化单克隆抗体偶合脂质体脑内寻靶提高基因转移效率,探讨将脂质体构建成更理想的脑内基因转移载体的方法。将MMA(抗Lex/SSEA-1单克隆抗体)衍生化后偶联于阳离子脂质体DOSPER上构建靶向性脂质体P-MMA-DOSPER,分别使用P-MMA-DOSPER与DOSPER包封pEGFP-C2(增强型绿色荧光蛋白质粒),分两组注入大鼠侧脑室后第1、3、7、14d取脑制备冰冻切片,抗nestin抗体免疫荧光染色,荧光显微镜下观察基因表达情况。结果显示两组大鼠侧脑室周围均可见绿色荧光蛋白(GFP)表达,各时间点P-MMA-DOSPER组的GFP+/nestin+细胞在nestin+细胞中比率均高于DOSPER组,差别具有统计学意义(P<0.01)。本研究表明P-MMA-DOSPER能够透过脑室管膜屏障高效安全地向脑内原位神经干细胞转移基因。This research tried improving the specificity and efficiency of gene transfection in gene therapy and tried making the liposome a better gene transfer vector to brain by use of the monoclonal antibody（ anti-Lex/SSEA-1 ）-mediated targeting of liposome. The derivatized monoclonal antibody was conjugated to the liposome DOSPER to form the targeting lipce, ome P-MMA-BOSPER. Then, the pEGFP-C2 encapsulated in P-MMA-DOSPER or DOSPER was injected into the lateral ventricle of SD rats respectively, and the brains were taken for frosted slice 1, 3, 7 or 14 days later. The expres- sion of GFP was observed under fluorescent microscope. There was a lot d expression of GFP around the lateral ventricle of rats in each group. But the indirect fluorescence antibody test showed the ratio of GFP＋/nestin＋ ceils to nestin＋ cells of every marking time point in the group of P-MMA-EOSPER was higher than the one in the group of DOSPER; the difference was found to be statistically significant （P 〈 0.01 ）. The results proved that the P-MMA-DOSPER can permeat the ependyma and can transfer gene into the nerve stem cells in vito safely and effectively.

关 键 词：侧脑室 脂质体 单克隆抗体 基因转染 绿色荧光蛋白 

分 类 号：Q782[生物学—分子生物学]