靶向ανβ3整合素超声造影剂的制备及其相对定量化的体外寻靶实验研究  

The Preparation of ανβ3 Integrin-targeted Contrast-Enhanced Ultrasound Agent and Relative Quantitative Targeted Assay in vitro

作　　者：赵捷颖 罗源 杨泽宏[2] 谢天骁 庄华[1] Zhao Jieying;Luo Yuan;Yang Zehong(Department of Ultrasound,West China Hospital of Sichuan University,Chengdu 610041,China;West China School of Basic Medical Sciences & Forensic Medicine,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院超声科,成都610041 [2]四川大学华西基础医学与法医学院,成都610041 [3]成都七中国际部高中,成都610041

出　　处：《华中科技大学学报（医学版）》2019年第4期382-387,共6页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.81101061);四川省科技计划资助项目(No.2017JY0074)

摘　　要：目的制备靶向ανβ3整合素的微泡超声造影剂,检测其物化特性并进行体外寻靶实验研究。方法采用生物素-链霉亲和素法制备靶向ανβ3整合素的微泡超声造影剂,检测其结合力与稳定性;制备细胞爬片,观察ανβ3整合素在受选细胞株的表达;相对定量地检测靶向微泡的体外寻靶潜能和结合特异性。结果靶向ανβ3整合素的微泡超声造影剂在液相的粒径均一,稳定性好,浓度高,抗体分子在微泡表面分布均匀。靶向ανβ3整合素的微泡较非靶向微泡有更高的结合微泡数,在每个HT29、诱导HUVECs、未诱导HUVECs和4T1细胞表面结合微泡数分别为(5.4±1.5)、(3.2±0.9)、(1.5±0.6)和(0.2±0.1),显示了良好的结合特异性和靶向性特征。结论成功制备了靶向ανβ3整合素超声造影剂,建立了相对定量化的体外寻靶实验方法,为进一步优化靶向微泡超声造影剂的体外寻靶策略提供了实验基础。Objective αvβ3 integrin-targeted microbubble was prepared to serve as contrast-enhanced ultrasound agent.The physicochemical characteristics were analyzed and targeted assays in vitro were performed.Methods αvβ3 integrin-targeted microbubble was prepared by biotin-streptavidin method and served as contrast-enhanced ultrasound agent.The related binding force and stability were tested.Cell plates were prepared and the distinct ανβ3 integrin expression was observed on cell surfaces.The relative quantitative in vitro targeting potential of ανβ3 integrin-targeted microbubble was detected and used to reflect the binding specificity.Results αvβ3 integrin-targeted microbubble had uniform size,stable shape,and high concentration in liquid phase.The biotinylated anti-CD51 or anti-CD61 molecules distributed evenly on the microbubble’s surface.Compared to the non-targeted microbubble,αvβ3 integrin-targeted microbubble showed significantly high binding microbubble number,and the number per cell was quantitatively calculated to be(5.4±1.5),(3.2±0.9),(1.5±0.6) and(0.2±0.1),respectively,on HT29,induced HUVECs,non-induced HUVECs and 4 T1 cell lines,which firmly demonstrated the binding specificity and relatively quantitative targeting features.Conclusion This assay successfully prepared a kind of αvβ3 integrin-targeted microbubble to serve as ultrasound-enhanced contrast agent and established relatively quantitative methodology to detect its targeting features in vitro.It provides the experimental research basis for optimizing the targeting strategies of αvβ3 integrin-targeted microbubble in vitro.

关 键 词：αvβ3整合素 造影剂 微泡 体外寻靶实验 

分 类 号：R319.2[医药卫生—基础医学]