靶向VEGFR-3载阿霉素脂质微泡超声造影剂体外显影及寻靶能力的实验研究  

Experimental study on enhancement effect and targeting study in vitro of VEGFR-3-targeted lipid microbubble ultrasound contrast agents containing adriamycin

作　　者：叶鸣[1,2] 王志刚[3] 周鸿[1,2] 牛诚诚[4] 陈昶宇[1,2] 刘莹[1,2] 

机构地区：[1]成都市第三人民医院 [2]重庆医科大学附属成都第二临床学院,四川成都610010 [3]重庆医科大学超声影像研究所,重庆400010 [4]中南大学湘雅二院超声科,湖南长沙410000

出　　处：《中国临床医学影像杂志》2015年第7期509-511,共3页Journal of China Clinic Medical Imaging

基　　金：四川省卫生厅科研课题项目(120536)

摘　　要：目的:制备一种靶向淋巴管内皮细胞载阿霉素脂质微泡超声造影剂,并考察其体外显影及寻靶能力。方法:制备生物素化抗VEGFR-3单克隆抗体,检测其生物素化程度;用机械振荡法,及生物素-亲和素桥连方式构建靶向VEGFR-3载阿霉素的脂质超声微泡。检测其一般特性;行体外超声,观察其显影效果;并在荧光下观察靶向载药微泡在体外与细胞的结合能力,与载药非靶向微泡做比较。结果:每个单抗分子平均可与9个生物素分子结合,所制备的载药靶向脂质微泡超声造影剂分散度佳,其平均粒径为1.66μm;体外超声显影,载药靶向脂质微泡呈高回声,回声强度随浓度及机械指数减小而降低;体外寻靶试验显示,该靶向载药脂质微泡较多并牢固的聚集在淋巴管内皮细胞表面。结论:成功制备的靶向VEGFR-3载阿霉素脂质微泡超声造影剂,有较高的包封率及载药率,其在体外有增强超声显影效果,并对淋巴管内皮细胞亲和力有较强的特异性。Objective： To prepare VEGFR-3-targeted lipid microbubble ultrasound contrast agents containing adfiamycin, and investigate their enhancement effect and affinity on human lymphatic endothelial cells in vitro. Methods： The biotinylated degree of prepared biotinylated monoclonal antibody VEGFR-3 was determined. The microbubbles containing adriamycin were prepared by mechanical vibration. Then the biotinylated antibody was attached to the surface of the microbubbles by avidin- biotin interaction. The physical property was determined and the enhancement effect in vitro was observed with ultrasound, The conjugation of targeted lipid microbubble ultrasound contrast agents containing adriamycin with human lymphatic endothe- lial cells was proved by light microscope and fluorescence microscope, with non-targeted microbubblcs served as control group. Results： About 9 biotin molecules were coupled to each antibody in average. The disposition of the prepared microbubbles was uniform size, the mean diameter was 1.66 μm. In vitro, the prepared microbubbles showed hyperechogcnicity in US imag- ing. In the targeting study in vitro, it was shown that the conjugation of the targeted microbubbles containing adriamycin with human lymphatic endothelial cells was tight while control group was negative. Conclusions： VEGFR-3-targeted lipid uhra- sound microbubbles contrast agents containing adriamycin are successfully prepared, with higher entrapment efficiency and drug-loading amounts, which have strong enhancement effect and targeting function in vitro.

关 键 词：内皮细胞 血管内皮生长因子受体3 淋巴管 造影剂 

分 类 号：R445.1[医药卫生—影像医学与核医学]