摘要
目的探讨超声靶向微泡破碎（UTMD）的物理靶向和纳米粒的生物靶向联合的双靶向方法细胞内递送siRNAs的效果。方法制备RGD和非RGD生物可降解纳米粒；L16（4^5）正交设计筛选超声、微泡、纳米粒的优化参数；实验分成优化参数组、RGD纳米粒组、非RGD纳米粒组和空白对照组；倒置荧光显微镜和流式细胞仪观察并检测分子探针Cy3标记的siRNAs在细胞内的摄取。结果RGD纳米粒组的细胞摄取率和荧光强度分别为（93．49±1．37）％和34．28±2．06，优化参数组的细胞摄取率和荧光强度分别为（88．33±1．24）％和30．59±3．93．两组比较差异均无统计学意义（P〉0．05）。非RGD纳米粒组的细胞摄取率和荧光强度分别为（71．24±2．80）％和18．39±0．90，优化参数组的细胞摄取率和荧光强度分别为（84．78±2．13）％和27．18±0．91，两组比较差异均有统计学意义（P〈0．05）。结论UTMD物理靶向和纳米粒生物靶向联合的双靶向方法不能提高siRNAs的细胞内递送：
Objective To investigate the intracellular delivery of siRNAs through the applications of ultrasound targeted microbubbles destruction（UTMD）and biodegradable nanoparticles carriers. Methods Preparation of nanoparticles with and without RGD sequences, parameters optimization via L16, （4^5 ） orthogonaI design, control experiments in groups of optimization, RGD targeted nanoparticles, non RGD nanoparticles and blank control, and determinations by inverted fluorescence microscope and flow cytometry were performed. Results The uptake and fluorescence intensity of PC-3 cells in group of RGD targeted nanoparticle was （93. 49 ± 1. 37）% and 34.28 ± 2.06 respectively, and that in group of optimization was （88.33 ± 1.24） % and 30.59±3.93 respectively（ P 〉0. 05）. Whereas the uptake and fluorescence intensity of PC 3 cells in group of non-RGD nanoparticles was（71.24 ± 2.80）% and 18.39 ± 0. 90 respectively, and that in group of optimization was （84.78 ± 2.13） % and 27.18 ±0.91 respectively（ P 〈0.05）. Conclusions The applications of UTMD with RGD targted nanoparticles cannot increase the intracellular delivery of siRNAs.
出处
《中华超声影像学杂志》
CSCD
北大核心
2011年第5期445-450,共6页
Chinese Journal of Ultrasonography
基金
国家自然科学基金项目（30772369/H1205）
上海市级医院适宜技术联合开发推广应用项目（SHDC12010221）
973国家重点基础研究发展计划（2010CB5299002010CB5299（12）
上海市科委纳米专项计划（1052nm05900）
关键词
超声检查
微气泡
基因治疗
Ultrasonography
Microbubbles
Gene therapy
