靶向MAdCAM-1超声造影剂的制备及其体外寻靶实验研究  

Preparation of targeted MAdCAM-1 ultrasound contrast agent and targeting study in vitro

作　　者：陈文颖[1] 郑凯[1] 黄敏[1] 

机构地区：[1]南京医科大学附属苏州医院超声科,江苏苏州215001

出　　处：《南京医科大学学报（自然科学版）》2014年第6期745-749,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省自然科学基金(BK20131150);苏州市科技发展计划(SYS201148)

摘　　要：目的:制备适用于炎症性肠病超声诊断并靶向MAdCAM-1的长循环脂膜超声造影剂,并鉴定其物理性状及体外寻靶能力。方法:采用超声破碎法制备长循环脂膜超声微泡,通过"生物素-亲和素"法桥接超声微泡与抗MAdCAM-1单克隆抗体(MECA-367)。对制备出的靶向微泡造影剂进行物理性状检测;通过光镜和激光共聚焦显微镜观察该靶向造影剂对TNF-α诱导的炎症性肠病细胞模型的结合能力。同法制备携同型对照抗体IgG2a微泡作对照。结果:制备的靶向超声微泡形态好且粒径较均匀,平均粒径(464.5±85.7)nm,Zeta电位(-19.3±2.5)mV。SVEC4-10细胞MAdCAM-1的表达与TNF-α刺激时间成正相关。并且,当TNF-α浓度达到20 ng/ml时,MAdCAM-1的表达达到峰值。体外寻靶试验表明,该靶向造影剂较多并牢固地黏附到表达MAdCAM-1的细胞周围;携IgG2a的同型对照组未见明显结合。结论:成功制备出桥连MECA-367的靶向长循环脂膜超声造影剂,该造影剂在体外对高表达MAdCAM-1的炎性细胞模型有较强的特异性亲和力;TNF-α诱导SVEC4-10细胞建立炎症性肠病细胞模型的方法简便易行。Objective：To prepare long-circulating lipid membrane ultrasound contrast agent suitable for the ultrasonic diagnosis of inflammatory bowel disease（IBD） and targeted mucosal addressin cell adhesion molecule-1（MAdCAM-1）,and determine the physical characteristics of the microbubbles and investigate their affinity for SVEC4-10 cells in vitro.Methods：The long-circulating lipid membrane microbubbles were prepared by sonic dispersion,and bridged the microbubbles and anti-murine MAdCAM-1monoclonal antibody（MECA-367） by ＂biotin-avidin＂.The physical traits of prepared targeted microbubble contrast agent were detected.Its ability to target to IBD cell model induced by tumor necrosis factor-α（TNF-α） was determined under light microscope and confocal laser scanning microscope,and isotype control antibody IgG2a microbubbles were used as control.Results：The targeted ultrasound microbubbles had a good shape and uniform particle size,with the average diameter of（464.5 ± 85.7）nm,and the surface Zeta potential was（-19.3 ± 2.5）mV.MAdCAM-1 expression on cultured SVEC4-10 cells was positively correlated with the time of TNF stimulation.In addition,when the concentration of TNF-a was 20 ng/ml,MAdCAM-1 expression reached its peak.The targeting study in vitro showed that many targeted ultrasound contrast agents adhered more firmly to the surrounding cells expressing MAdCAM-1,while the control group had no significant binding.Conclusion：Bridging MECA-367 long-circulating lipid membrane ultrasound contrast agents were successfully prepared.The targeted ultrasound contrast agents can bind to the cell model of high expression MAdCAM-1 effectively in vitro,and the method of establishing IBD cell model on TNF-stimulated SVEC4-10 cells is simple and feasible.

关 键 词：靶向 超声造影剂 MADCAM-1 炎症性肠病 细胞模型 

分 类 号：R445.1[医药卫生—影像医学与核医学]