聚乙烯亚胺增强Pluronic介导的BALB/c小鼠骨骼肌原位基因传递和表达效率  

Polyethyleneimine Enhances in situ Gene Delivery and Expression in Skeletal Muscles of BALB/c Mice in the Pluronic-Mediated System

作　　者：何雨桐 孙哲 王刚 HE Yutong;SUN Zhe;WANG Gang(Engineering Research Center in Biomaterials, Sichuan University, Chengdu 610064, China)

机构地区：[1]四川大学生物材料工程研究中心,成都610064

出　　处：《四川动物》2018年第6期620-627,共8页Sichuan Journal of Zoology

基　　金：国家自然科学基金项目(31370972)

摘　　要：目的在Pluronic L64介导的BALB/c小鼠骨骼肌基因传递体系中正确引入聚乙烯亚胺(PEI)以增强基因表达效率。方法以生理盐水组、质粒DNA (p DNA)组和Pluronic L64/p DNA (L/D)组为对照组,各比例PEI/p DNA与Pluronic L64混合物(L/P/D-0. 5、L/P/D-3、L/P/D-10)为实验组,将各组样品注射到小鼠胫骨前肌中。7 d后取组织,检测报告基因表达(β-半乳糖苷酶和荧光素酶)和肌肉组织病变情况。通过尾静脉注射考察L/P/D-0. 5组的体内急性毒性。最后,通过动态光散射仪和原子力显微镜测定L/P/D-0. 5组中PEI/p DNA复合物的粒径、电位及形貌。结果 1)报告基因表达评估:L/P/D-0. 5组外源基因表达效率最高;随着N/P增加,几乎检测不到外源基因的表达。2)生物相容性评估:无论原位注射还是尾静脉注射,L/P/D-0. 5组均没有显示出组织病变,而其他比例的L/P/D组出现了不同程度的组织学病变。3)物理特征表征:虽然在L/P/D-0. 5组中DNA未被完全压缩,但PEI/p DNA复合物已拥有良好的纳米粒径和形态结构,且表面电位为负。结论 L/P/D-0. 5体系能安全有效地增强外源基因在骨骼肌内的表达。Objective To correctly introduce cationic polyethyleneimine (PEI) into the Pluronic L64-mediated gene de- livery system of mice skeletal muscle to enhance the efficiency of gene expression. Methods In this study, different PEI samples were transferred into mice tibialis anterior muscles via intramuscular injection. The saline, plasmid DNA (pDNA) and Pluronie If34/pDNA (L/D) were used as control groups, and the mixtures of PEI/pDNA with different N/P ratios (L/P/D-0. 5, L/P/D-3, and L/P/D-10) and Pluronie If94 were used as experimental groups. Muscle tissues were taken out at 7 days after injection, and then the report gene expression of fl-galactosidase and lueiferase and the potential pathological changes of muscle tissues were measured. In addition, the in vivo acute toxicity experiment was further determined by injecting L/P/D-0. 5 through tail vein. Finally, the particle size, zeta potential and morphology of PEIZpDNA complexes in L/P/D-0. 5 system were detected by dynamic light scattering instrument and atomic force microscope (AFM). Results 1 ) The L/P/D-0. 5 group showed the highest report gene expression ; As N/P ratio increasing, the report gene expression could hardly be detected. 2) The result of bioeompatibility assessment showed that no tissue lesion was observed in L/P/D-0. 5 group irre- spective of muscular injection or intravenous injection, while other ratios of L/P/D showed histologic lesions with varying de-grees. 3) PEI/pDNA complexes were spherical shape with nanometer size, and this might contribute to the process of gene de- livery in skeletal muscle-based system, although pDNA was not fully compressed in L/P/D-0. 5 group. Conclusion The I.,/P/D-0. 5 system can safely and effectively enhance the expression of exogenous genes in skeletal muscle.

关 键 词：BALB/C小鼠 骨骼肌基因传递 聚乙烯亚胺 PLURONIC L64 

分 类 号：Q95-336[生物学—动物学]