基因转染的pH敏感脂质体的最优处方和制备工艺的研究

Resarch on the optimized recipe and preparation conditions of pH-sensitive liposomes of gene transfection with orthogonal design methods

目的 寻找用于基因转染的pH敏感脂质体的最佳处方和制备工艺。方法 采用正交设计法 ,考虑制备的pH值、组成摩尔比、制备方法三因素 ;即 1个四水平因素和 2个二水平因素混合状态。选用L16(4 1·2 12 )正交设计表安排试验 ,各水平组合条件下 ,通过基因转染实验 ,以基因转染荧光测定值作为观测指标 ,实验重复 4次。结果 在基因与脂质体重量比为 (1∶8)时 ,组成大豆磷脂与DC 胆固醇摩尔比为 (6∶4)和在超声法制备的特定条件下 ,pH5 .4与pH7.4基因转染荧光测定值之间差别非常显著 ;均值都较大 ,但是以 pH5 .4时基因转染荧光测定值均值最大。 结论 基因转染荧光测定值最高的脂质体为在 pH5 .4、组成大豆磷脂与DC 胆固醇摩尔比为 (6∶4)时采用超声法制备的脂质体。

OBJECTIVE To search the optimized recipe and preparation conditions of pH-sensitive liposomes.METHODS The orthogonal design methods were adopted through gene transfection test.The fluorescence intensity was used as the index of gene transfection.RESULTS The fluorescence intensity of gene transfection was the highest when the liposomes were constituted by soybean lecithin and DC- cholesterol (6∶4,molar ratio) at pH 5.4 condition and were prepared by using ultrasonication.The effects of different factors on the preparation of pH-sensitive liposomes were studied.There was significant difference in the mean fluorescence intensity between pH5.4 and pH7.4.The mean fluorescence intensity of gene transfection was the biggest at pH5.4.CONCLUSION The optimized preparation conditions of pH-sensitive liposomes with soybean lecithin and DC-cholesterol (6∶4, molar ratio) which had the highest fluorescence analysis value of gene transfection were acquired using ultrasonication methods at pH 5.4.