Institute of Ultrasound Imaging, Department of Ultrasonography, the Second Affiliated Hospital of Chongqing Medical University, No. 76, Lin Jiang Road, YuZhong District, ChongQing City, 400010, China.
Acad Radiol. 2009 Dec;16(12):1457-65. doi: 10.1016/j.acra.2009.06.018. Epub 2009 Sep 24.
Ultrasound-targeted microbubble destruction is a promising technology for the targeted gene delivery. The purpose of the present study is to prepare a novel lipid ultrasound microbubble-carrying gene and transactivating transcriptional activator (Tat) peptide and to investigate its transfection effect in vivo.
Lipid ultrasound microbubbles were prepared using mechanical vibration, and the appearance, distribution, concentration, diameter, and zeta potential of the lipid ultrasound microbubbles were measured. The efficiencies of the microbubble carrying gene and Tat peptide were investigated using the fluorospectrophotometer. Contrast-enhanced ultrasonography was performed on normal rabbits to observe the duration and intensity of enhancement in myocardium. Quantitative analysis was detected using the DFY Ultrasound Image Analyzer. Transfection in vivo was performed using the CGZZ ultrasound gene transfection instrument. The expression of enhanced green fluorescent protein in the organs was observed using confocal laser scanning microscope.
The diameter of the lipid microbubbles carrying gene and Tat was (2.3 +/- 0.4) mum, the concentration was (3.1 +/- 0.4) x10(9)/mL, and Zeta potential was (2.0 +/- 0.1) mV. The gene encapsulation efficiency of the lipid ultrasound microbubbles was 32%, and the Tat encapsulation efficiency was 35%. In vivo experiment showed that lipid ultrasound microbubbles could enhance the echo intensity and transfection efficiency.
Lipid microbubbles containing gene and Tat peptide can be used as a new vehicle for gene transfection.
超声靶向微泡破坏是一种有前途的靶向基因传递技术。本研究的目的是制备一种新型脂质超声微泡载基因和转激活转录激活剂（Tat）肽，并研究其体内转染效果。
采用机械振动法制备脂质超声微泡，测量脂质超声微泡的外观、分布、浓度、直径和zeta 电位。采用荧光分光光度计研究微泡携带基因和 Tat 肽的效率。对正常兔进行超声造影检查，观察心肌增强的持续时间和强度。采用 DFY 超声图像分析仪进行定量分析。采用 CGZZ 超声基因转染仪进行体内转染。利用共聚焦激光扫描显微镜观察器官中增强型绿色荧光蛋白的表达。
携带基因和 Tat 的脂质微泡的直径为（2.3 ± 0.4）μm，浓度为（3.1 ± 0.4）×10（9）/mL，Zeta 电位为（2.0 ± 0.1）mV。脂质超声微泡的基因包封效率为 32%，Tat 的包封效率为 35%。体内实验表明，脂质超声微泡可以增强回声强度和转染效率。
载有基因和 Tat 肽的脂质微泡可以作为一种新的基因转染载体。
