双叶酸靶向超声纳泡的制备及其体外寻靶能力的实验研究  

Experimental study on preparation and targeting in vitro of nanobubbles with two-fold amount of folate

作　　者：段素娟[1] 郭鲁[1] 时丹丹[1] 商蒙蒙 孟冬[1] 孙霄[1] 史本康[2] 李杰[1] Duan Sujuan;Guo Lu;Shi Dandan;Shang Mengmeng;Meng Dong;Sun Xiao;Shi Benkang;Li Jie(Department of Ultrasound,Qilu Hospital,Shandong University,Jinan 250012,China)

机构地区：[1]山东大学齐鲁医院超声科,济南250012 [2]山东大学齐鲁医院泌尿外科,济南250012

出　　处：《中华超声影像学杂志》2018年第11期995-999,共5页Chinese Journal of Ultrasonography

基　　金：国家自然科学基金资助项目(81771843);山东省重点研发计划(2017GSF18107).

摘　　要：目的制备新型双叶酸靶向纳泡[(FOL)2-NBs],探讨其物理表征及与高表达叶酸受体的乳腺癌MCF-7细胞的靶向结合能力。方法合成新型叶酸靶向材料DSPE-PEG2000-AD-(FOL)2,机械振荡法制备新型双叶酸纳泡(FOL)2-NBs,检测其粒径及增强显影效果,体外寻靶实验检测其对叶酸受体(folatereceptor,FR)过表达乳腺癌MCF-7细胞的靶向结合能力。结果核磁共振氢谱(1HNMR)提示DSPE-PEG2000-AD-(FOL)2缀合物的成功合成,纯度为90%。新型(FOL)2-NBs光镜下呈圆形,平均粒径为(286.87±22.96)nm,具备良好的增强显影效果。体外寻靶实验提示,与传统的单叶酸靶向纳泡相比,新型(FOL)2-NBs介导的MCF-7靶向结合能力显著提高。结论成功制备新型双叶酸纳泡(FOL)2-NBs,其与叶酸受体过表达的肿瘤细胞靶向结合能力更强,有望作为叶酸受体过表达肿瘤血管外分子诊疗的理想造影剂。Objective To investigate the selective targeting ability of a novel folate-modified nanobubbles with two-fold amount of folate [(FOL)2-NBs] . Methods DSPE-PEG2000-AD-(FOL)2with two-fold of folate per DSPE-PEG2000 chain was synthesized and then tested by 1 H nuclear magnetic resonance (1H NMR) . The novel (FOL)2-NBs was prepared using the mechanical shaking method based on lipid-stabilized perfluoropropane . The bubble size was measured by Malvern laser particle size analyzer and the contrast enhancement ability was also detected with imaging machine using a self-made agarose mold . The experiment of selective targeting ability was also carried out in human breast cancer MCF-7 cell with over-expression of folate receptor ( FR) using fluorescence activated cell sorting ( FACS) . Results The result of 1H NMR proved that DSPE-PEG2000-AD-( FOL )2was successfully synthesized ,and the purity reached up to 90% . The novel prepared ( FOL) 2-NBs showed superior contrast enhancement ability with a particle size of ( 286.87±22.96) nm . Compared with the conventional NBs ,the novel ( FOL) 2-NBs exhibited improved selective cellular targeting ability proven by FACS . Conclusions A novel nanobubble with improved selective targeting ability is successfully prepared and shows great potential in extravascular imaging and curation in FR overexpressed tumors .

关 键 词：超声造影 纳泡 叶酸受体 乳腺癌 

分 类 号：R445.1[医药卫生—影像医学与核医学]