侧脑室注射免疫脂质体介导GFP基因脑内表达时程

目的探索以免疫脂质体携带基因侧脑室注射,透过脑室管膜屏障脑内寻靶。方法以衍生化的抗 Lex／SSEA—1单克隆抗体(P—MMA)偶联阳离子脂质体DOSPER构建免疫脂质体P-MMA-DOSPER。P-MMA- DOSPER包封绿色荧光蛋白质粒(pEGFP-C2)。注入SD大鼠侧脑室,于术后1、3、7、14、21d取脑,冷冻切片nestin抗体免疫荧光染色,荧光显微镜观察。结果注射脂质体质粒复合物一侧侧脑室周围均有大量GFP表达,并可见于对侧侧脑室及第三脑室周围,术后1d、21d GFP表达明显低于其他时间点,差异有统计学意义(P<0．01);3d、7d、14d GFP ／nestin 细胞在nestin 细胞中比例均高于1d、21d,差别均有统计学意义(P<0．01)。结论免疫脂质体P- MMA-DOSPER能够透过脑室管膜屏障高效安全的向脑内原位神经干细胞转移基因。

Expression of GFP gene in brain mediated by immunoliposome injected into lateral ventricle

Objective To make gene transferin targeting in brain by injecting gene encapsulated in immunoli-posome into lateral ventricle. Methods The derivatized monoclonal antibody was conjugated to the liposome DOSPER to form the immunoliposome P-MMA-DOSPER. Injecting the pEGFP-C2 encapsulated in P-MMA-DOSPER into lateral ventrcle of SD rats and take the brains 1,3,7,14,21d later. Observing the expression of GFP through the fluorescent microscope. Results There was a lot of expression of GFP around the lateral ventricle and the third ventricle. The expression of GFP on 1 and 21d after geng transferin was lower than the one of 3,7,14d. The indirect fluorescence antibody test showed the GFP /nestin cell's ratio of nestin cells in the group of 3.7, 14d after gene transferin was higher than the one in the group of 1 and 21d,the diference was found to be statiticlly significant (P<0. 001). Conclusions The immunoliposome P-MMA-DOSPER can permeat the blood brain barrier and transfer gene into the nerve stem cells in vivo safely and effectively.