亚微米级荧光脂质微泡的制备及其肝脏成像的实验研究  

Preparation of DiO-labeled microbubbles and assessment of its liver enhancement

作　　者：乔璐[1] 高文宏[1] 张莉[1] 梁红敏[2] 李露[1] 朱梅[2] 李佩倞[1] 刘政[1] 

机构地区：[1]第三军医大学新桥医院超声科,重庆400037 [2]昆明医学院第一附属医院超声科,昆明650032

出　　处：《第三军医大学学报》2013年第9期858-861,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30860269);云南省应用基础研究计划面上项目(C0620056Q)~~

摘　　要：目的制备荧光标记的脂质微泡,检测一般物理性质、荧光标记情况以及评价在兔肝脏的显影效果。方法用二甲基亚砜(dimethyl sulfoxide,DMSO)溶解细胞膜绿色荧光分子探针(3,3 dioetadeeyloxaearboeyanineperehlorate,DiO)后,抽取少量溶解液加至脂质冻干粉内与溶媒液相溶,全氟丙烷置换瓶内空气,最后机械振荡仪高速振荡制成带有荧光标记的脂质微泡,检测其微泡的一般物理性质、荧光标记情况;随机选取6只健康新西兰大白兔,雄性,体质量2.0～2.5 kg,经兔耳缘静脉团注荧光微泡后动态观察微泡在兔肝实质的显影效果,手术切取部分肝脏,冰冻切片后在荧光显微镜下观察荧光素在肝脏内的分布情况。结果荧光标记的脂质微泡形态呈圆形,大小均一,微泡浓度范围(4～7)×109/mL,微泡粒径为(429±119)nm,激光共聚焦显微镜下微泡外壳被绿色光圈环绕。兔肝脏造影可见持续饱满的增强显影效果,荧光显微镜下肝脏内荧光素面积分布较少。结论荧光标记的脂质微泡大小均一,粒径大小达到进入肺循环标准,在兔肝脏内显影效果显著。Objective To prepare DiO-labeled microbubbles, and to investigate their physicochemical property, fluorescent labeling conditions and the enhance effects for liver contrast imaging. Methods The DiO-labeled microbubbles were prepared with a freeze-dried powder and mixed Dimethyl sulfoxide （DMSO） dissolved DiO. Then we replaced the air in the bottles with perfluoropropane. Finally the DiO-labeled micro- bubbles were made by high speed mechanical agitation. The physicochemical property and fluorescent labeling conditions of DiO-labled rnicrobubbles were investigated. Six healthy male New Zealand white rabbits which weighted between 2.0 and 2.5 kg were injected through auricular vein with the DiO-labeled microbubbles. Then we dynamically observed the microbubbles enhanced ultrasound images of the rabbit liver parenchyma. Fluorescence microscopy was used to observe the deposition of green fluorescein in a small portion of frozen section liver tissues. Results DiO-labeled microbubbles were round in shape and uniform in size, with a concentration of （4 -7） x 109/mL amt a size of 429 4- l l9 nm. The outer shell of DiO-labeled microbubbles was green under confoeal laser scanning microscope. Liver imaging using DiO-labeled mierobubbles showed significant and persistent enhancement. Little fluorescein was distributed in the liver under fluorescence micro- scope. Conclusion DiO-labeled microbubbles are uniform in size and can access the pulmonary circulation. They significantly enhance the liver images in contrast enhanced ultrasound.

关 键 词：DiO标记物 脂质微泡 超声 肝脏 

分 类 号：R445.1[医药卫生—影像医学与核医学] R943[医药卫生—诊断学]