人表皮生长因子受体2靶向新型高分子超声造影剂的制备及体外寻靶实验研究  

Reparation of human epithelial growth factor receptor 2-targeted polymer ultrasound contrast agents with liquid perfluorocarbons and in vitro experiment

作　　者：柯青兰[1] 王志刚[1] 何子朋[1] 岳媛媛[1] 李攀[1] 宋卫香[1] 

机构地区：[1]重庆医科大学超声影像学研究所重庆医科大学附属第二医院超声科,400010

出　　处：《中华超声影像学杂志》2014年第1期66-70,共5页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金（81227801,81130025,81110219,81371579）;重庆市高校创新团队计划（KJTD201303）

摘　　要：目的制备一种新型血管外靶向超声造影剂——包裹液态氟碳的人表皮生长因子受体2（HER2）靶向高分子纳米球，并观察其体外寻靶、显影能力。方法首先采用单乳化法制备包裹液态氟碳的高分子PLGA纳米球（PFH—PLGA），然后通过碳二亚胺法将靶向HER2的特异性抗体trastuzumab连接到纳米球表面，制成靶向纳米球（PFH—PLGA—tra）；检测其一般表征；免疫荧光法检测trastuzumab与纳米球连接情况；体外寻靶实验观察PFH—PLGA-tra与人乳腺癌细胞MCF-7的结合情况；体外加热法观察液态氟碳相变情况；体外观察纳米球增强超声显影效果。结果靶向液态氟碳造影剂平均粒径为（261±28）nm；免疫荧光法显示trastuzumab被成功地连接到了纳米球上；体外寻靶实验示，该靶向造影剂较多并牢固地聚集在表达HER2的MCF-7细胞表面，对照组未见造影剂和MCF-7的结合；体外热相变实验示，较多纳米球转变为微泡；体外显影实验示，造影剂回声强度和灰度均较双蒸水明显增强（P〈0．05）。结论通过单乳化法和碳二亚胺法成功制备了靶向HER2的液态氟碳高分子超声造影剂，其粒径较小，靶向能力较强，具有相变潜能，有望成为HER2阳性肿瘤血管外显影、治疗的理想造影剂。Objective To prepare a novel extravascular human epithelial growth factor receptor 2 （HER2）-targeted polylactic-co-glycolie acid （PLGA）-COOH ultrasound contrast agent with liquid perfluorocarbons（PFH-PLGA-tra）, and to observe it＇s general properties and itrs effects on ultrasound imaging in vitro. Methods Polymeric nanocapsules of liquid perfluorocarbons were prepared by the single emulsion technique and conjugated with trastuzumab monoclonal antibody by EDC/NHS. The general characteristics were observed. The conjugation was demonstrated by immunofluorescence and the targeting performance of the agent was checked in human MCF-7 cells line in vitro. The encapsulation of liquid fluorocarbons was detected by heating in vitro. The effects on ultrasound imaging were observed in vitro. Results The mean diameter of agents was （261 ＋ 28） nm. The targeted nanocapsules were positive in immunofluorescence. In the targeted study,it was shown that a number of targeted nanocapsules conjugated with MCF-7 cells in vitro. Droplet-to-bubble transition experiment in vitro showed that there were no nano/ microbubbles formed by heating in control groups at 80~C, while there were a lot of nano/microbubbles appeared in nanocapsules formulations at 80 ~C. The agents presented a good ultrasound imaging in vitro. Conchusions The HER2-targeted polymeric nanocapsules of liquid perfluorocarbons are successfully prepared,which may become a novel extravascular targeted ultrasound contrast agents.

关 键 词：超声检查 微气泡 纳米技术 液态氟碳 

分 类 号：R96[医药卫生—药理学]