PEG-PEI/ROCK-Ⅱ-siRNA复合物物理性能和转染C17.2神经干细胞效率的研究  

The characteristics of PEG-PEI/ROCK-Ⅱ-siRNA complexes and its transfection efficiency in C17.2 neural stem cells

作　　者：刘云云[1] 刘中霖[1] 王勇[2] 程度[2] 梁嫣然[1] 温小军[1] 刘军[1] 胡竞扬[1] 刘大成[3] 杨幸怡[3] 

机构地区：[1]中山大学孙逸仙纪念医院神经内科,广东广州510120 [2]中山大学化学与化学工程学院,广州510275 [3]中山大学中山医学院,广州510080

出　　处：《解剖学研究》2012年第6期401-405,共5页Anatomy Research

基　　金：国家自然基金(81000466);中山大学2012年国家大学生创新项目(201210558100)

摘　　要：目的探索PEG-PEI/ROCK-Ⅱ-siRNA复合物的物理性能及转染C17.2神经干细胞的效率。方法设计合成PEG-PEI共聚物基因载体,与ROCK-Ⅱ-siRNA进行复合;用凝胶阻滞电泳实验、粒径与电位的测定观察PEG-PEI/ROCK-Ⅱ-siRNA复合物的复合效果;采用透射电镜观察PEG-PEI/ROCK-Ⅱ-siRNA复合物的形态学;通过流式细胞仪技术定量检测复合物对C17.2神经干细胞的转染效果。结果 PEG-PEI/ROCK-Ⅱ-siRNA复合物在N/P=10时完全复合。随着N/P比的增大,复合物的粒径变小而电位增大。在N/P=50时,透射电镜观察到复合物为球形,很好的分散性,平均粒径约为100 nm。PEG-PEI/ROCK-Ⅱ-siRNA复合物在N/P=50时转染C17.2神经干细胞的转染效率最高,为(70.24±5.21)%。结论 PEG-PEI基因载体可以很好的与ROCK-Ⅱ-siRNA复合。PEG-PEI/ROCK-Ⅱ-siRNA复合物对于C17.2神经干细胞具有良好的转染效率。Objective To envaluate the characteristics of PEG-PEI/ROCK- Ⅱ-siRNA complexes and its transfection effi- ciency in C 17.2 neural stem cells. Methods The compound effect of PEG-PEI/ROCK- Ⅱ -siRNA complexes were tested by gel retardation assay, particle size and zeta potential analysis. Morphology of PEG-PEI/ROCK- Ⅱ-siRNA complexes were detected by transmission electron microscopy. The transfeetion efficiency were measured by flow cytometry. Results PEG-PEI/ROCK-Ⅱ -siRNA complexes were completely eompouned at N/P=10. When the N/P ratio increased, the size of the PEG-PEI/ROCK-Ⅱ-siRNA com- plexes decreased, but the zeta potential increased. Transmission electron microscopy demonstrated that the PEG-PEI/ROCK-Ⅱ-siR- NA complexes （N/P=50） were spherical and well dispersed. The mean diameter of the complexes was approximately 100 nm. At N/ P ratio=50, the transfection efficiency of PEG-PEI/ROCK- Ⅱ -siRNA complexes was reached （70.24 e 5.21 ）%. Conclusion PEG- PEI was a good gene carrier for delivering ROCK- Ⅱ -siRNA. PEG-PEI/ROCK- Ⅱ -siRNA complexes displayed excellent transfeetion efficiency in C17.2 neural stem cells.

关 键 词：聚乙二醇-聚乙烯亚胺 ROCK-Ⅱ RNA干扰 纳米医学 

分 类 号：R749.16[医药卫生—神经病学与精神病学]