CAJ | 학술논문

目的研究聚乙二醇—聚乙烯亚胺(PEG-PEI)作为非病毒纳米载体介导小干扰RNA(siRNA)体外转染C17.2 NSCs的效果。方法采用自行设计合成的PEG-PEI与靶向Nogo受体复合体基因的siRNA形成复合物，通过粒径与电位的测定、凝胶阻滞电泳实验等方法观察PEG-PEI/siRNA纳米复合物的表征及复合效果。以脂质体复合体系Lipofectamine 2000/siRNA为对照，采用流式细胞仪检测不同N/P比(PEG-PEI中氮原子和siRNA中磷原子的摩尔比)的PEG-PEI/siRNA复合物对NSCs的转染效率。结果 PEG-PEI和siRNA形成粒径为纳米级别的复合物，随着N/P比增大，复合物的粒径逐渐减小，而表面电位逐渐增大。凝胶阻滞电泳实验表明siRNA与PEG-PEI可以通过静电相互作用而稳定结合。流式细胞仪检测细胞转染率后发现，N/P=15时，PEG-PEI/siRNA复合物转染率最高，可达(78.72±8.18)％。结论 PEG-PEI是一种有良好发展前景的非病毒型siRNA转运载体，对神经干细胞基因治疗具有潜在的应用价值。
목적 연구취을이순—취을희아알(PEG-PEI)작위비병독납미재체개도소간우RNA(siRNA)체외전염C17.2 NSCs적효과。 방법 채용자행설계합성적PEG-PEI여파향Nogo수체복합체기인적siRNA형성복합물，통과립경여전위적측정、응효조체전영실험등방법관찰PEG-PEI/siRNA납미복합물적표정급복합효과。이지질체복합체계Lipofectamine 2000/siRNA위대조，채용류식세포의검측불동N/P비(PEG-PEI중담원자화siRNA중린원자적마이비)적PEG-PEI/siRNA복합물대NSCs적전염효솔。 결과 PEG-PEI화siRNA형성립경위납미급별적복합물，수착N/P비증대，복합물적립경축점감소，이표면전위축점증대。응효조체전영실험표명siRNA여PEG-PEI가이통과정전상호작용이은정결합。류식세포의검측세포전염솔후발현，N/P=15시，PEG-PEI/siRNA복합물전염솔최고，가체(78.72±8.18)％。 결론 PEG-PEI시일충유량호발전전경적비병독형siRNA전운재체，대신경간세포기인치료구유잠재적응용개치。
Objective To explore the effect of polyethylene glycol-polyethyleneimine (PEG-PEI) serving as a non-viral vector in delivering small interfering RNA (siRNA) into C17.2 neural stem cells (NSCs) in vitro. Methods Complexes of PEG-PEI and siRNA targeting Nogo receptor were prepared, and their characterizations were estimated by measurements of particle size and zeta potential,and the complex abilities of PEG-PEI/siRNA complexes were observed by gel retardation assay. In addition, with liposome complex system (Lipofectamine 2000/siRNA) as positive control, the transfection efficiency of PEI-PEG/siRNA complexes at different N/P ratios (cationic nitrogen/siRNA phosphate molar ratio) was detected by flow cytometry. Results The siRNA molecules were condensed by PEG-PEI to form nanoseale complexes. As the proportion of N/P ratio enhancing, the surface potential of nanoparticles gradually increased and the particle sizes of PEI-PEG/siRNA complexes showed a decreasing trend. Gel retardation electrophoresis suggested that siRNA could be fully composited with PEG-PEI as a result of the coulombic foree between them. Meanwhile, flow cytometry experiments revealed that the transfection efficiency of PEG-PEI mainly depended on N/P ratios of the nanoparticles,and the highest one was obtained at N/P=15 ([78.72±8.18)]％). Conclusion PEG-PEI might be a prospective candidate for siRNA delivery system, which enjoys its value in NSC gene therapy.