郭 信,赵笑天,吴小雨,刘俊超,刘晓兵.新型PEI衍生物的体外表征及对巨噬细胞的转染效果研究[J].现代生物医学进展英文版,2019,19(14):2620-2625. |
新型PEI衍生物的体外表征及对巨噬细胞的转染效果研究 |
Characterization of Novel PEI Derivatives and Transfection Effects on Macrophages |
Received:March 07, 2019 Revised:March 30, 2019 |
DOI:10.13241/j.cnki.pmb.2019.14.004 |
中文关键词: GPEI 巨噬细胞 基因转染 |
英文关键词: GPEI Macrophage Gene transfection |
基金项目:上海市浦江人才计划项目(14PJ1406100) |
Author Name | Affiliation | E-mail | GUO Xin | Department of Vascular Surgery, The Ninth People's Hospital of Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China | guo_xin_2015@163.com | ZHAO Xiao-tian | School of Pharmacy Shanghai Jiaotong University, Shanghai, 200240, China | | WU Xiao-yu | Department of Vascular Surgery, The Ninth People's Hospital of Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China | | LIU Jun-chao | Department of Vascular Surgery, The Ninth People's Hospital of Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China | | LIU Xiao-bing | Department of Vascular Surgery, The Ninth People's Hospital of Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China | |
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中文摘要: |
摘要 目的:以PEI1.8kDa为基础连接戊二醛合成新型可降解PEI衍生物GPEI后,研究GPEI包裹质粒转染巨噬细胞后对巨噬细胞合成蛋白及表型的影响。方法:合成GPEI后,检测了GPEI质粒聚合物的粒径及电位,并通过透射电镜观察聚合物形态,通过免疫荧光、WB、RT-PCR、Elisa及Transwell实验检测巨噬细胞RNA、蛋白、终产物水平变化及表型改变。结果:GPEI能够包裹质粒,形成100 nm左右带有正电荷的聚合物,WB、RT-PCR结果表明GPEI转染较PEI25kDa有优势(P<0.05),巨噬细胞转染后6天持续分泌PGI2(P<0.05)。结论:以PEI1.8kDa为基础合成的GPEI能够包裹PTGIS质粒并将其转染进巨噬细胞后对巨噬细胞的表型有明显影响。 |
英文摘要: |
ABSTRACT Objective: Synthesizing a novel degradable PEI derivative(GPEI), and studying the protein level and phenotype of macrophage transfected by GPEI-plasmid. Methods: The particle size and potential of GPEI-plasmid were detected by Zeta potentiometer. The morphology of GPEI-plasmid was observed by transmission electron microscopy. The RNA, protein, end products and phenotypic changes were detected by immunofluorescence, WB, RT-PCR, Elisa and Transwell experiments. Results: Plasmid coated GPEI was positive charge and about 100 nm. GPEI transfection is superior to commercial PEI25kDa(P<0.05), and PGI2 was secreted for 6 days by transfected macrophage(P<0.05). Conclusion: The synthesized GPEI can encapsulate the PTGIS plasmid and successfully transfect it into macrophages, which has a significant effect on the phenotype of macrophages. |
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