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【目的】制备血管生成素-2小干扰RNA(Ang-2siRNA)质粒纳米微粒,并观察其基因转染能力和基因沉默效果。【方法】应用分子克隆的方法构建Ang-2siRNA质粒,将其与聚乳酸、O-羧甲基壳聚糖用水/油/水(W/O/W)双乳化溶剂蒸发法制备纳米微球,扫描电镜观察其形态和粒径。然后转染原代人脐带静脉内皮细胞, 观察纳米Ang-2siRNA微粒干扰Ang-2基因表达的效果及其细胞保护功能。【结果】扫描电镜观察到Ang- 2siRNA纳米微球呈球形和椭球形,粒径150—200 nm,大小分布均匀,包封完整,分散性良好。有较强转基因能力,良好的RNA干扰效果和血管内皮细胞保护功能。【结论】成功制备纳米Ang-2siRNA微粒。并证实其载基因能力和RNA干扰效果,对血管内皮细胞损伤因素有良好保护作用,为进一步研究心肌梗死的血管修复机制奠定了坚实的基础。
【Objective】 To prepare Ang-2 siRNA plasmid nanoparticles and observe its gene transfection ability and gene silencing effect. 【Method】 Ang-2 siRNA plasmid was constructed by molecular cloning. Nanospheres were prepared by double emulsion solvent evaporation of polylactic acid and O-carboxymethyl chitosan with water / oil / water (W / O / W) Scanning electron microscopy to observe its morphology and particle size. Then primary human umbilical vein endothelial cells were transfected to observe the effect of nano-Ang-2 siRNA particles interfering with Ang-2 gene expression and cell protection. 【Results】 Scanning electron microscopy showed that the Ang-2 siRNA nanospheres were spherical and ellipsoidal with the size of 150-200 nm. The size distribution was uniform, the encapsulation was complete and the dispersion was good. Strong transgene ability, good RNA interference effect and vascular endothelial cell protection function. 【Conclusion】 Nanoparticles of Ang-2 siRNA were successfully prepared. And confirmed its gene-carrying capacity and RNA interference effect on vascular endothelial cell injury factor has a good protective effect for further study of myocardial infarction vascular repair mechanism has laid a solid foundation.