纳米血管生成素-2小干扰RNA质粒制备及功能研究

Encapsulation of Ang-2 Small Interfering RNA Shuttle Vector in Poly Lactic Acid /O-carboxy Methylated Chitosan Nanospheres and Its Functional Investigation

【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 small interfering RNA (siRNA) shuttle vector nanosphere and evaluate both its gene-delivery ability and its gene-silence effect. [Methods] The nanosphere was prepared by the solvent evaporation technique with Poly (lactic acid)/O-carboxymethylated and Ang-2 siRNA shuttle vector. The microscopic morphology of the nanosphere was examined by scanning electron microscope. The gene silence effect of nanosphere for transfected human umbilical vein endothelium cells (HUVEC) was observed. [Results] The nanospheres were spherical or ellipse shape. Their diameters were in the range of 150-200 nm. Its gene-delivery and RNA interfering ability were perfect, [Conclusion] We synthesized Ang-2 (siRNA) shuttle vector nanosphere successfully and proved both its gene-delivery ability and its RNAi effect. It laid the foundation vascular renovation about myocardial infarction.