ASODN-Protamine-HSA-PLGA纳米粒的构建及体外细胞核靶向的初步研究

目的 研究新型非病毒载体ASODN-Protamine-HSA-PLGA纳米粒的制备方法及其对体外细胞核靶向的可能性.方法 用鱼精蛋白硫酸盐(protamine sulfate)和人血清白蛋白(HSA)缩合反义寡核苷酸(ASODN), 再以聚丙交酯乙交酯(PLGA)作为载体材料, 采用双乳化溶媒蒸发法制备载ASODN-Protamine-HSA-PLGA纳米粒(ASODN-P/H-PLGA-NP).用透射电镜观察其形态;用激光粒度仪测定其粒径、多分散性和zeta电位;用两步法测定其包封率;用MTT法测定其毒性;用激光共聚焦显微镜观察其进入人喉颈鳞癌细胞(Hep-2)和靶向细胞核的能力.结果 制备的纳米粒形态圆整,大小均匀,平均粒径为128 nm,多分散系数(PDI)为0.234,平均zeta电位为-23.3 mV,平均包封率为78.45%.PLGA 纳米粒对ASODN-P/H可起到较好的保护作用,ASODN-P/H-PLGA-NP能够顺利的进入Hep-2细胞,并具有较强的细胞核靶向作用.结论 ASODN-P/H-PLGA-NP是一种具有细胞核靶向潜力的非病毒载体系统.

Preparation of ASODN-Protamine-HSA-PLGA Nanoparticles and Initial Evaluation of Their Nucleus Targeting Property in vitro

Objectives To develop a novel non-viral gene delivery system-SODN-Protamine-HSA-PLGA(ASODN-P/H-PLGA-NP)and investigate its nucleus targeting potential in vitro.MethodsASODN-P/H-PLGA-NP was prepared by mixing the protamine sulfate and HSA.Then the PLGA nanoparticles were prepared using double-emulsion evaporation technique,followed by addition of ASODN to the prepared P/H complex.The morphology of ASODN-P/H-PLGA-NP was observed by transmission electron microscopy.The diameter,PDI,and surface charge of ASODN-...