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目的载小鼠白介素18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乙二醇/乙交酯丙交酯作为载体材料,采用双乳化溶剂蒸发法制备了载聚阳离子-DNA复合物纳米粒(polymer-polycationic peptide-DNA,PPD),用透射电镜观察该多聚复合物的形态,用激光粒度仪测定其粒径和Zeta电位,琼脂糖电泳分析抗核酸酶和抗超声的能力,用Westernblot检测在HepG2肝癌细胞中的表达情况。结果制得的纳米粒,形态圆整,大小均匀,平均粒径(155±2.97)nm,平均Zeta电位为(-27.31±2.02)mV,平均包封率91.12%,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,细胞转染表达效率也显著优于裸质粒。结论PPD纳米粒是基因治疗的临床应用中一种很具前景的多聚复合物载体系统。
AIM: To study the properties and expression of mouse interleukin - 18 (IL - 18) recombinant plasmid nanoparticles. Methods Polyethylene glycol / glycolide lactide, a non-toxic biodegradable macromolecule material, was used as a carrier material to prepare polymer-polycationic peptide- DNA, PPD). The morphology of the polyplexes was observed by transmission electron microscopy. The particle size and zeta potential were measured by laser particle sizer. The ability of anti-nuclease and anti-ultrasound to analyze by agarose electrophoresis was analyzed by Western blot in HepG2 hepatoma cells The expression of the situation. Results The nanoparticles prepared were round and uniform in size with an average diameter of (155 ± 2.97) nm and an average zeta potential of (-27.31 ± 2.02) mV, with an average entrapment efficiency of 91.12% The ability of plasmids to resist ultrasonic shear and nuclease degradation was also significantly improved compared with naked plasmid. Conclusion PPD nanoparticle is a promising multimeric complex system in the clinical application of gene therapy.