Annexin V微泡对凋亡组织寻靶的试验研究  

Experimental Study on Targeting of Apoptotic Tissue By Annexin V Microbubbles

作　　者：李梦琴 朱梅[1] 冯煜然 范旭 Li Mengqin;Zhu Mei;Feng Yuran;Fan Xu(Department of Ultrasound,the First Affiliated Hospital of Kunming Medical University,Kunming,Yunnan 650032,China)

机构地区：[1]昆明医科大学第一附属医院超声科,昆明市650032

出　　处：《中国超声医学杂志》2020年第4期359-361,共3页Chinese Journal of Ultrasound in Medicine

基　　金：云南省应用基础研究重点项目(No.2016FA031);云南省科技厅云岭技术产业人才项目;云南省卫计委医学学科带头人项目(No.D-201648)。

摘　　要：目的制备Annexin V靶向微泡,评价其理化性质,并进行初步体外寻靶试验探究。方法生物素-亲和素桥接法制备Annexin V靶向微泡,粒径分析仪、Malvern电位仪评价其理化性质,免疫荧光染色法评估靶向配体结合状况,用荧光显微镜观察其形态、大小及分布状态。建立人甲状腺未分化癌裸鼠模型,对其进行微波消融,诱导肿瘤组织凋亡。随机分组,A组为普通微泡,B组为Annexin V抗体预饱和+靶向微泡,C组为Annexin V靶向微泡,比较3组微泡与消融后凋亡肿瘤组织的结合情况。结果成功制备Annexin V靶向微泡,微泡形态大小一致,理化性质稳定;A组和B组,基本未见微泡与肿瘤组织切片稳定结合,而C组见Annexin V微泡与组织稳定结合。结论采用生物素-亲和素桥接法制成Annexin V靶向微泡;且在体外能与凋亡肿瘤组织稳定结合,即成功寻靶,从而实现识别检测凋亡组织的功能。Objective To prepare Annexin V targeted microbubbles(AVMBs),evaluate their physicochemical properties,and conduct preliminary targeting search in vitro.Methods The AVMBs were prepared by biotin-avidin bridge method.The physicochemical properties were evaluated by particle size analyzer and Malvern potentiometer.The binding status of targeting ligands was evaluated by immunofluorescence staining.The morphology,size and distribution of microbubbles were observed by fluorescence microscopy.A model of thyroid cancer was established on nude mice by human thyroid undifferentiated cancer cell line,and the tumor was ablated by microwave to induce apoptosis of tumor tissue.Randomly grouped,compared the combination of different microbubbles with apoptotic tumor tissues after ablation in group A,B,and C with apoptotic tumor tissue after ablation.Group A is common microbubbles,group B is Annexin V antibody presaturation+targeted microbubbles,and group C is AVMBs.Results Annexin V targeted microbubbles were successfully prepared with uniform size and stable physicochemical properties;In group A and group B,there was almost no microvesicles and tumor tissue sections were stably combined,while Group C showed AVMBs and tissue stable binding.Conclusions AVMBs were prepared by biotin-avidin bridge method.AVMBs can stably bind to apoptotic tumor tissues in vitro,that is,target is successfully sought,so as to realize the function of identification and detection of apoptotic tissue.

关 键 词：ANNEXIN V 靶向微泡 检测 肿瘤 凋亡 

分 类 号：R445.1[医药卫生—影像医学与核医学] R736.1[医药卫生—诊断学]