| 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].,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 |
| 投稿时间：2019-03-07 修订日期：2019-03-30 |
| 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） |
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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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