何勤, 张志荣, 刘戟, 徐超群. 载TK基因聚丙交酯乙交酯纳米粒的性质及表达研究J. 药学学报, 2004, 39(4): 285-287.
引用本文: 何勤, 张志荣, 刘戟, 徐超群. 载TK基因聚丙交酯乙交酯纳米粒的性质及表达研究J. 药学学报, 2004, 39(4): 285-287.
HE Qin ZHANG Zhi-rong, LIU Ji, XU Chao-qun, . Study on the preparation of TK-gene nanoparticles and its expressionJ. Acta Pharmaceutica Sinica, 2004, 39(4): 285-287.
Citation: HE Qin ZHANG Zhi-rong, LIU Ji, XU Chao-qun, . Study on the preparation of TK-gene nanoparticles and its expressionJ. Acta Pharmaceutica Sinica, 2004, 39(4): 285-287.

载TK基因聚丙交酯乙交酯纳米粒的性质及表达研究

Study on the preparation of TK-gene nanoparticles and its expression

  • 摘要: 目的载pEGFP-TKAFB重组质粒纳米粒的性质及其表达研究。方法以无毒的可生物降解的高分子材料聚丙交酯乙交酯作为载体材料,采用双乳化溶媒蒸发法制备了载pEGFP-TKAFB重组质粒纳米粒,考察其形态学及包封率,琼脂糖电泳分析抗核酸酶抗超声的能力,MTT法测定GCV对细胞的抑制率,流式细胞仪测定报告基因EGFP的表达。结果制得的纳米粒,形态圆整,大小均匀,平均粒径(72±12) nm,平均包封率91.25%,质粒制成纳米粒后提高了质粒对抗超声剪切及核酸酶降解的能力,细胞转染效率也显著优于裸质粒。结论质粒DNA制成纳米粒可进一步研究基因药物的给药系统。

     

    Abstract: AimTo prepare TK-gene nanoparticles and investigate its expression. MethodsBiodegradable and biocompatible polymer polylactic-co-glycolic acid (PLGA) was used to prepare recombinant plasmid ppo nanoparticles by double-emulsion evaporation technique. The characteristics of the nanoparticles including morphology, entrapment efficiency was investigated. The expression of TK gene was also investigated by MTT assay, which could determine the dying cells after the addition of gancyclovir (GCV). The enhanced green fluorescent protein (EGFP) expression in human hepatocarcinoma SMMC-7221 cells and human normal parenchymal Chang liver cells were assessed by flow cytometric analysis. ResultsThe resulting plasmid-nanoparticles had regular spherical surface and a narrow particle size with a mean diameter of (72±12) nm, The average entrapment efficiency was 91.25%, the enhanced transfection efficiency and ability protecting plasmid DNA from degraded by nuclease or sonication due to nanoparticles encapsulation. ConclusionDNA-nanoparticles need further study as gene delivery system.

     

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