中文摘要：

目的制备靶向卵巢癌的纳米超声微泡造影剂，鉴定其理化特性并观察其体外寻靶能力。方法采用冷冻干燥超声法制备非靶向纳米超声微泡造影剂（N-N.Mb），通过生物素一亲和素桥连法，将生物素化的促黄体生成素释放激素（1uteinzing hormone-releasing hormone，LHRH）与N-N-Mb连接，制备靶向卵巢癌的纳米超声微泡造影剂（LHRH-N-Mb）。观察微泡镜下形态以及室温下保存时间。zeta检测仪检测粒径范围、表面电位，流式细胞术检测二抗分别与LHRH-N-Mb、N-N-Mb结合率，光镜下观察LHRH-N-Mb与LHRH受体过表达的人卵巢癌OVCAR-3细胞结合情况。结果N-N-Mb、LHRH-N-Mb两种微泡成功制备，两者外观圆整，分布均匀，粒径大小分别为（372±45）、（5114±64）nm，集中于360nm与508nm，两者粒径大小比较有统计学差异（P〈0.05）。两种微泡电位分别为（-14.5±0.5）、（-14.6±0.8）mV。室温下保存16d后，LHRH-N-Mb理化特性与刚制备时无统计学差异（P〉0.05）。二抗与LHRH-N-Mb、N-N-Mb结合率分别为75.6％、0.83％，两者比较有统计学差异（P〈0.05）。光镜下OVCAR.3细胞周围可见LHRH-N-Mb围绕或粘附，呈花环样结构，这种靶向结合能被生物素化的LHRH抗体预先阻断。结论通过冷冻干燥超声法和生物素.亲和素桥连法，可成功制备靶向卵巢癌的纳米超声微泡造影剂，此微泡粒径小、稳定性高，且能体外靶向高效结合人卵巢癌OVCAR-3细胞。

英文摘要：

Objective To prepare luteinising-hormone releasing hormone （LHRH） nanoliposomal microbubbles specifically targeting ovarian cancer cells and characterize their physiochemical properties and in vitro targeting ability. Methods Lyophilization/sonication method was used to prepare non-targeting nanoliposomal microbubble （N-N-Mb）. By using the biotin-avidin bridge method, conjugated biotinylated LHRH antibodies to N-N-Mb generated biotinylated LHRH nanoliposomal microbubble （LHRH-N-Mb） targeting specifically ovarian cancer cells. The morphology and shelf life of microbubbles was detected by light microscopy, and the particle size range and the surface potential were measured by zeta detector. The binding affinity of the secondary antibody with LHRH-N-Mb or N-N-Mb was determined by flow cytometry. The binding of LHRH-N- Mb to human OVCAR-3 ovarian cancer cells was detected by light microscopy. Results The rounded and uniformly distributed N-N-Mb and LHRH-N-Mb were successfully generated. The particle size was 372 ± 45 nm with a mean of 360 nm for N-N-Mb, and 511± 64 nm with a mean of 508 nm for LHRH-N-Mb. There was significant difference in the size between the 2 groups （ P 〈 0. 05 ） , while the surface potential of the 2 microbubbles remained no significant difference （ - 14.5 ± 0.5 and - 14.6±0.8 mV, P 〉 0.05 ）. After being kept at room temperature for 16 d, no significant difference of the LHRH-N-Mb＇ s phvsiochemical orooerties was detected compared with that of freshly prepared microbubbles. The secondary antibody binding rate of LHRH-N-Mb or N-N-Mb was 75.6% or 0.83% respectively, with significant difference （ P 〈 0.05 ）. Moreover, the rosette formation surrounding OVCAR-3 cells was observed under light microscope after cells were incubated with LHRH-N-Mb while pre-incubated biotinylated LHRH antibody blocked the rosette formation. Conclusion The LHRH nanoliposomal microbubbles specifically targeting ovarian cancer cells are successfully prepared through biotin-avidin mediation