固体脂质纳米粒作为核酸疫苗载体的效应研究

Transfection efficiency study on LDL like cationic SLN as carrier of DNA vaccine delivery

文章模拟低密度脂蛋白制作阳离子固体脂质纳米粒,并评价其作为核酸疫苗载体的实用性。利用改进的溶剂乳化法成功制备LDL样固体脂质纳米粒(SLN)。SLN组成成分为胆固醇油酸酯(47%,W/W),甘油三酯(2%,W/W),DOPE(9%,W/W)、未酯化胆固醇(12%,W/W)、DC-胆固醇(30%,W/W)。结果表明,固体脂质纳米粒的平均大小为(98.96±0.24)nm,表面zeta电位为(+177±14.14)m V。相比聚乙烯亚(PEI),SLN在同等浓度下表现出相当微弱的细胞毒性。在最佳质量比值条件下,SLN和p EGFP-C2载体复合物(8.4??1)对体外培养细胞的转染能力得到大幅度改善,相比单用p EGFP-C2载体转染时强约10倍。以(p Shuttle-H1-荧光素酶sh RNA)/SLN复合物处理转荧光素酶基因的人肺腺癌细胞PC9时,复合物成功抵达细胞内并通过si RNA干扰途径显示出实质性的荧光素酶基因沉默现象。试验合成的LDL样阳离子固体脂质纳米粒颗粒大小均匀、表面正电荷稳定、细胞毒性甚微,可与p EGFP-C2真核表达载体结合,有效转染到细胞内。

To investigate synthesized LDL like cationic solid lipid nanoparticles（SLN） as a carrier of DNA vaccine delivery. SLN were prepared by modified solvent-emulsification method and were tested as a carrier of DNA vaccine delivery. The SLN consisted of 47% （W/W） cholesteryl oleate, 2% （W/ W） triglyceride, and 12% （W/W） cholesterol, and 9% （W/W） L-alpha-dioleoyl phosphatidyl-ethanolamine （DOPE）, and 30% （W/W） 3/3-[N-（N＇, N＇-Dimethyl amino- ethane）-carbamoyl] Cholesterol Hydrochloride （DC-chol）. Results showed that the SLN had a mean diameter of （98.96 ± 0.24） nm and a surface zeta potential value of （＋ 177±14.14） mV. The SLN exhibited much lower cytotoxicity than that of polyethylenimine （PEI, 25K） when has been used in the same concentration. Under an optimal weight ratio （8.4： 1） of DC-chol of SLN and pEGFP-C2 plasmid conjugate, the complexes exhibited substan- tially enhancing gene expression than naked plasmid. When administered into PC9 human lung cancer cells expressing luciferase, （pShuttle-Hl-luciferase shRNA）/SLN complexes were successively transferred into the inner cell region with showing significant gene silencing. The synthesized LDL like cationic SLN had uniform particle size and stable positive charge on their surface, low cytotoxicity and could well transfect into eukaryotic cell when combined with pEGFP-C2 plasmid.