CAJ | 학술논문

目的探讨一种新型可负载siRNA微泡的制备方法,以Skov-3细胞为实验对象,验证其体外siRNA转染的效能.方法利用声振法制备表面带负电的微泡,与聚合物修饰的siRNA孵育,获得携siRNA的微泡；用动态光散射法和透射电镜观察其形态结构；用流式细胞仪和倒置荧光显微镜检测体外转染Skov-3细胞的效果.结果携siRNA的微泡表面粗糙,直径为(332.0±20.4)nm；低频超声辐照后,细胞转染效率为(57.3±2.5)％,siRNA主要分布于胞浆.结论成功制备负载siRNA的微泡,在低频超声辐照后具有较高的体外转染效能,为下一步基因治疗研究提供siRNA递送系统.
목적 탐토일충신형가부재siRNA미포적제비방법,이Skov-3세포위실험대상,험증기체외siRNA전염적효능.방법 이용성진법제비표면대부전적미포,여취합물수식적siRNA부육,획득휴siRNA적미포；용동태광산사법화투사전경관찰기형태결구；용류식세포의화도치형광현미경검측체외전염Skov-3세포적효과.결과 휴siRNA적미포표면조조,직경위(332.0±20.4)nm；저빈초성복조후,세포전염효솔위(57.3±2.5)％,siRNA주요분포우포장.결론 성공제비부재siRNA적미포,재저빈초성복조후구유교고적체외전염효능,위하일보기인치료연구제공siRNA체송계통.
Objective To explore a method for preparing siRNA-loaded nanobubbles (siRNA-NBs) with a special structure and determine the siRNA transfection efficiency in vitro.Methods siRNA-NBs were formed by hetero-assembly of nanobubbles and polymeric siRNA-micelles.Dynamic light scatter and transmission electron microscope were employed to determine the novel structure of siRNA-NBs.Meanwhile,flow cytometery and inverted fluorescence microscope were used to investigate the in vitro siRNA transfection efficiency of siRNA-NBs with low frequency ultrasound exposure.Results The siRNANBs possessed uneven surfaces with the diameter of (332.0 ± 20.4)nm.After low-frequency ultrasound exposure,the in vitro siRNA transfection efficiency of siRNA-NBs was (57.3 ± 2.5)％,with the widely cytoplasmic siRNA distribution.Conclusions The successfully preparation of siRNA-NBs with high transfection efficiency in vitro indicates that the novel ultrasound contrast agent are of great potential in siRNA targeting delivery and gene therapy.