携P选择素单抗靶向微泡在高剪切应力下体外血栓超声分子成像的效果  

In Vitro Efficacy of Microbubbles Targeted to P-selectin on Ultrasound Molecular Imaging of Thrombus in High-shear Stress

作　　者：黄瑞珠[1] 杨莉[2] 胡广全[1] 廖旺军[3] 伍巍兰[1] 吴爵非[1] 滕中华[1] 原野[1] 宾建平[1] 

机构地区：[1]南方医科大学南方医院心内科,广州市510515 [2]南方医科大学南方医院药学部,广州市510515 [3]南方医科大学南方医院肿瘤科,广州市510515

出　　处：《中国超声医学杂志》2010年第11期966-969,共4页Chinese Journal of Ultrasound in Medicine

基　　金：国家"863"计划项目(No.2006AA02Z478);国家自然科学基金(No.30870722)

摘　　要：目的制备携P-选择素单抗的靶向超声微泡(MBp),通过体外流动腔模型评价其在高剪切应力下行血栓超声分子成像的效果。方法采用"亲和素-生物素"桥接法制备MBp和同型对照微泡(MB),应用自制琼脂糖流动腔模型模拟体内高剪切应力血流环境,将MBp和MB随机先后注入琼脂糖模型内,与小鼠血栓孵育30min后,PBS液15cm/s流速不间断冲洗血栓,分别在冲洗2、4、6、8和10min时行对比超声检查并测量声强度(VI)值。免疫组化评价小鼠血栓的P-选择素表达。结果冲洗前MBp组和MB组血栓VI值无明显差异(P>0.05);冲洗后VI值在各时间点MBp组均较MB组大(P<0.05)。冲洗10min后MBp组血栓仍有可视性对比增强,而MB组血栓在冲洗2min后已无可视性对比增强。免疫组化显示血栓表面有明显的P-选择素表达。结论以P-选择素为靶标的MBp在高血流剪切应力下与血栓的靶向结合稳定可靠,可用于动脉血栓的超声分子成像。Objective To prepare microbubbles targeted to P-selectin （MBp）and assess its efficacy on ultrasound molecular imaging of thrombus in high-shear stress in vitro. Methods The biotinylated anti-mice P-selectin （MBp）or isotype control antibody （MB）was conjugated to the biotinylated lipid-mierobubbles via a streptavidin bridge. An agarose flow-chamber model was homely made as an imitation of the system of arterial blood flow. Thrombus was developed in vitro and fixed in flow chamber model. After 30 min incubation with equivalent MB or MBp,the thrombus was washed with the continuous flow of PBS solution in high speed （15cm/s）. Contrast enhanced ultrasound imaging of thrombi was performed at 2,4,6,8 and 10 min of the washing and the video intensity （VI）of thrombi was measured. Results Before the washing, there was no significant difference in the VI between both MBp and MB group （P〉0.05）. At each time points of the washing,the VI in the MBp was bigger than that in the MB （P〈0.05）. The visual enhancement of thrombus was still noted in the MBp with the washing of 10 min, while it was not seen even at 2 min in the MB. The surface of thrombus was positive for P-selectin. Conclusions MBp provides a good targeting performance to the P-selectin of thrombus in high-shear flow and it could be used to achieve ultrasound molecular imaging of artery thrombus.

关 键 词：靶向超声微泡 对比超声 P-选择素 血栓 

分 类 号：R943[医药卫生—药剂学]