新型靶向纳米材料作为基因载体的评估  

Evaluation of a novel targeted nanoparticles as gene vector

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

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510150 [2]广州医科大学附属第四医院神经内科,广东广州511447 [3]广州医科大学附属第二医院神经内科,广东广州510000

出　　处：《现代预防医学》2015年第13期2405-2408,共4页Modern Preventive Medicine

基　　金：广东省自然科学基金;批文号:S2013010011760

摘　　要：目的合成可降解性多聚赖氨酸-聚乙烯亚胺-聚乙二醇(PLL-PEG-PEI)并研究其负载质粒DNA(p DNA)的能力,比较所形成纳米材料/DNA复合物在体外对细胞的毒性大小及其转染效率,为进一步体内基因治疗做好准备。方法以PEI 25 k D为对照,并化学方法合成可降解性PLL-PEG-PEI,检测所形成纳米材料/DNA复合物对PBMC的细胞毒性,并对细胞进行转染,分别用流式细胞仪、荧光素酶基因表达水平和倒置荧光显微镜检测转染效果。结果PLL-PEG-PEI复合p DNA后,采用MTT法检测出复合物在PBMC细胞中的细胞毒性,并证实PLL-PEG-PEI细胞毒性比常用的PEI 25ku低,在PBMC细胞中,转染效率从整体来看,N/P比=15时,PLL-PEG-PEI萤光素酶基因表达水平最高,类似的,通过流式细胞仪和倒置荧光显微镜更直观的检测了转染效果,表明PLL-PEG-PEI对PBMC细胞有靶向效果且在N/P比为15时靶向效果最好。结论此研究结果为多聚赖氨酸靶向基因治疗神经系统自身免疫性疾病寻找行之有效的基因载体提供了理论和实践基础,可进一步于动物体内进行联合基因治疗中枢神经免疫疾病。Objective The PLL-PEG-PEI which was degradable was synthesized to enhance its ability of transporting p DNA. The transfection efficiency and toxicity were compared for its advanced gene therapy. Methods The PLL-PEG-PEI which was degradable was synthesized according to the PEI 25 k D. The toxicity and transfection efficiency were tested in PBMC cell with MTT assay, EGFP/fluorescent image, reporter assay and flow cytometry. Results These derivatives reduced the cytotoxicity of PEI 25 ku in PBMC cell. Compared with PEI 25 ku, the transfection efficiency increased. Reporter assay and flow cytometry showed that PLL-PEG-PEI/p DNA complexes exhibited higher transgene activity than that of PEI/p DNA in PLL-receptor. Conclusion The results indicated that PLL-PEG-PEI might be a promising candidate for gene delivery with the characteristic of good biocom patibility and relatively high gene transfection efficiency.

关 键 词：纳米材料 PLL-PEG-PEI 体外基因转染 

分 类 号：R115[医药卫生—公共卫生与预防医学]