超声微泡造影剂介导pEGFP-N1转染小鼠角膜的实验研究  

SonoVue and ultrasound mediated pEGFP-N1 transfection to mouse cornea in vivo study

作　　者：伍瑛[1] 杜联芳[1] 陈永东[2] 王慧萍[3] 王丰[3] 

机构地区：[1]上海交通大学附属第一人民医院超声科,200080 [2]上海交通大学附属第一人民医院眼科 [3]上海交通大学附属第一人民医院中心实验室

出　　处：《中华超声影像学杂志》2008年第4期350-353,共4页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金（30772369）,上海市自然科学基金（05ZR14087）

摘　　要：目的探讨微泡造影剂声诺维联合超声介导绿色荧光蛋白质粒转染小鼠角膜的转染效率和安全性。方法小鼠眼前房分别注射生理盐水、质粒＋生理盐水、质粒＋造影剂、脂质体＋质粒等，以50Hz，2W／cm。的脉冲波超声辐照质粒＋生理盐水和质粒＋造影剂眼10min。术后第1d、第3d、第7d、第14d和第21d荧光体视镜观察眼表EGFP表达出现情况，冰冻切片荧光显微镜观察阳性细胞的分布，病理切片观察组织有无损伤。结果造影剂联合超声辐照组小鼠眼表出现绿色荧光蛋白的弥漫性表达，明显高于单纯超声辐照组和脂质体转染组，其余对照各组眼表均未见绿色荧光。第3d时的病理切片各组均无异常。结论声诺维联合超声辐照在体能安全、有效地介导基因转染眼组织。Objective To investigate the efficiency and safety of transfection green fluorescent protein plasmid to mouse cornea mediated by SonoVue and ultrasound. Methods Saline, plasmid ＋ saline, plasmid ＋ SonoVue and liposome ＋ plasmid were injected respectively to mouse eye anterior chamber. Then the mouse eye of plasmid ＋ saline and plasmid ＋ SonoVue injection group were exposed to pulse wave ultrasound under 50 Hz pulse repetition frequence,2 W/cm^2 intensity and 10 minutes duration time. Fluorescence stereomicroscope was used to observe the expression of EGFP in the eye at the 1st day,3rd day,Tth day, 14th and 21st day after injection. Two mice were taken randomly from each group and were sacrificed at the 3rd day after injection. Their eyes were enucleated and made into frozen coronal sections. And fluorescence microscopy was performed to observe the type and distribution of EGFP positive cell. Tissue damage was observed in pathological section. Results EGFP was expressed over the ocular surface in SonoVue and ultrasound group, and it obviously higher than only ultrasound exposure group and liposome group. The expression of EGFP was not detected over the ocular surface in other groups. Pathological sections were not found any difference in each group. Conclusions SonoVue and ultrasound can successfully and safely transfer gene to ocular in vivo study.

关 键 词：超声检查 造影剂 基因转移技术 角膜 

分 类 号：R686[医药卫生—骨科学]