靶向纳米材料作为大鼠单个核细胞基因载体的研究  

A Study of Targeted Nano Materials as PBMCs Gene Vector

作　　者：董亚贤[1] 尧慧燕 梁兵[3] 钟高贤[2] 刁芳明[2] 石红婷[2] 

机构地区：[1]广州医科大学附属第一医院神经内科,广东广州510000 [2]广州医科大学附属第四医院神经内科,广东广州510000 [3]广州医科大学附属第二医院神经内科,广东广州510000

出　　处：《赣南医学院学报》2016年第3期343-346,共4页JOURNAL OF GANNAN MEDICAL UNIVERSITY

基　　金：广东省自然科学基金(批文号:S2013010011760)

摘　　要：目的:探讨纳米材料聚乙二醇-聚乙烯亚胺(PEG-PEI)作为基因载体的可行性研究,研究其负载质粒DNA(p DNA)的能力,分析所形成纳米材料/p DNA复合物的特性,以及体外对细胞的毒性大小及其转染效率。方法:以PEI为对照,合成PEG-PEI,采用MTT法检测所形成纳米材料/DNA复合物对大鼠单个核细胞(PBMC)的细胞毒性,再采用倒置荧光显微镜及流式细胞仪检测转染效果。结果:N/P<10时,PEG-PEI和PEI的存活率分别为82.7%及81.3%(P﹥0.05);N/P=10时,PEG-PEI是81%,PEI为59%(P<0.05);N/P﹥10时,PEG-PEI的细胞存活率比PEI低(P<0.05)。N/P=10时,用倒置荧光显微镜观测PEG-PEI较PEI表达的绿色荧光多,荧光强度大(P<0.05);同时利用流式细胞仪检测PEG-PEI和PEI的转染效率,分别为28.9%和12.5%(P<0.05)。结论:纳米材料PEG-PEI具有强的负载能力,转染效率高,细胞毒性低的优点,为作为多发性硬化基因载体的可行性奠定了实践基础。Objective： To investigate the feasibility of targeted nanoparticles as gene vector,we synthesized polyethyleneglycol polyethylenimine（ PEG-PEI）,and studied its capability of carrying plasmid DNA,analyzed its characterization as nanoparticle/DNA complex,assessed its cytotoxicity in vitro and measured its transfection efficiency. Methods： using PEI as control,we first developed PEG-PEI,and tested its cytotoxicity to the rat PBMCs via MTT assay. The transfection efficiency of the nanoparticle/DNA complex was also measured by EGFP immunofluorescence and flow cytometry. Results： When the N/P 10,the viability of PEG-PEI and PEI transfected cells were 82. 7% and 81. 3% respectively（ P﹥ 0. 05）; when the N/P = 10,the viability of PEG-PEI transfected cells was 81%,but that of PEI transfected cells was59%（ P〈0. 05）; similarly,when the N/P ﹥ 10,MTT assay results showed that the toxicity of the PEG-PEI was lower than PEI（ P〈0. 05）. The EGFP intensity of PEG-PEI（ N/P = 10） was much higher than PEI（ N/P = 10,P〈0. 05）assessed by inverted microscope fluorescence observation method; the flow cytometry results also showed that the transfection efficiency of PEG-PEI（ N/P = 10） and PEI（ N/P = 10） was 28. 9% and 12. 5%,respectively（ P〈0. 05）. Conclusion： The nanopaticle（ PEG-PEI） exhibited lower toxicity and higher transfection efficiency,and therefore is the more suitable transfection carrier for gene therapy for multiple sclerosis.

关 键 词：纳米材料 PEG-PEI 体外基因转染 

分 类 号：R285.5[医药卫生—中药学]