携ApoPep-1纳米微泡靶向成像评价活体内肿瘤凋亡的实验研究  

ApoPep-1 Peptide Nanobubbles Targeted Imaging for Assessment of Tumor Apoptosis in vivo:An Experimental Study

作　　者：赵萍[1] 马晓菊[1,2] 杨恒丽 袁丽君 段云友[1] ZHAO Ping;MA Xiaoju;YANG Hengli;YUAN Lijun;DUAN Yunyou(Department of Ultrasonic Medicine,Tangdu Hospital,the Fourth Military Medical University,Xi’an 710038,China;不详)

机构地区：[1]第四军医大学唐都医院超声诊断科,陕西西安710038 [2]临潼康复疗养中心健康管理部特诊科,陕西临潼710600 [3]西安医学院第二附属医院门诊超声科,陕西西安710038

出　　处：《中国医学影像学杂志》2020年第12期901-906,共6页Chinese Journal of Medical Imaging

基　　金：国家自然科学基金(81201136);空军军医大学唐都医院苗子人才资助基金。

摘　　要：目的以ApoPep-1为分子探针设计一种新的靶向纳米微泡,研究其在活体肿瘤组织内靶向凋亡细胞的成像能力,探讨其用于早期抗肿瘤疗效评价的意义。材料与方法多肽固相合成法合成生物素化ApoPep-1,并分析其分子量和纯度;将生物素化ApoPep-1以“生物素-亲和素-生物素”系统与生物素化纳泡连接得到靶向纳米微泡(ApoPep-1-NB),并分析其粒径大小和稳定性;采用超声仪检测其体外显影效果;顺铂治疗裸鼠建立肿瘤凋亡模型,超声造影观察其靶向成像能力;体外免疫荧光验证分析。结果ApoPep-1-NB粒径为(588.61±19.89)nm,且具有较好的稳定性和体外显影效果。ApoPep-1-NB在肿瘤组织中(60 s:107.56±3.96;120 s:116.06±2.88;180 s:118.68±2.08)较AV-NB(60 s:73.39±0.60;120 s:76.20±0.86;180 s:63.14±1.09)表现出更好的造影增强效果(P<0.01),且造影增强时间更长,尤其能清楚地显示内部高增强的凋亡区。免疫荧光实验与超声造影结果一致。结论本研究设计的ApoPep-1-NB稳定性更好,在肿瘤组织凋亡区的靶向成像能力更强,为早期肿瘤疗效评价提供了更有力的实验证据。Purpose To prepare ApoPep-1-nanobubbles,research the imaging ability of targeting to apoptosis of tumor in vivo and discuss the significance of the early evaluation of therapeutic effects.Materials and Methods The bio-ApoPep-1 was synthesized by the method of solid phase synthesis of polypeptides,and then the molecular weight and purity were analyzed.The ApoPep-1-NBs were prepared by the“biotin streptavidin-biotin bridging chemistry”.The size and stability of the ApoPep-1-NBs were measured by a particle analyzer.The echogenicity of ApoPep-1-NBs in vitro was measured using an ultrasound machine.Tumor apoptosis model was established,the tumor-bearing nude mice were treated with cisplatin.Contrast-enhanced ultrasound was performed to observe the ability of targeted ultrasound imaging in vivo.Finally,immunofluorescence examination was performed to confirm the results of ultrasound imaging.Results The size of ApoPep-1-NBs was(588.61±19.89)nm,it was relatively stable and had a feature of high echogenicity in vitro.In addition,ApoPep-1-NBs had higher echogenicity(60 seconds:107.56±3.96;120 seconds:116.06±2.88;180 seconds:118.68±2.08)in vivo and significantly longer enhancement inside the tumors after chemotherapy compared with AV-NBs(60 seconds:73.39±0.60;120 seconds:76.20±0.86;180 seconds:63.14±1.09,P<0.01).Moreover,the highly enhanced apoptotic region in the tumor tissue was also clearly demonstrated.The immunofluorescence confirmed the result of the targeted contrast-enhanced ultrasound.Conclusion We developed a novel ApoPep-1-NBs with stable particle size,which has a good ability of targeted imaging for apoptosis in vivo.This study provides an alternative tool for the early evaluation of cancer management.

关 键 词：超声检查 造影剂 分子成像 微泡 小鼠 裸 疾病模型 动物 细胞凋亡 ApoPep-1 

分 类 号：R73[医药卫生—肿瘤] R445.1[医药卫生—临床医学]