载Survivin-siRNA和蛇床子素阳离子脂质体纳米粒的制备工艺优化  

Optimization of preparation process of cationic liposome nanoparticles containing Survivin siRNA and osthol

作　　者：孙起超 白皓天 李娅兰 孟健男[2] 郭雨薇 王锐[2] SUN Qi-chao;BAI Hao-tian;LI Ya-lan;MENG Jian-nan;GUO Yu-wei;WANG Rui(The First Affiliated Hospital of Heilongjiang University of Chinese Medicine;Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院 [2]黑龙江中医药大学,黑龙江哈尔滨150040

出　　处：《海南医学院学报》2022年第1期6-12,共7页Journal of Hainan Medical University

基　　金：哈尔滨市应用技术研究与开发项目(2016RAQXJ197)。

摘　　要：目的:利用脂质体可同时装载水溶性成分和脂溶性成分的特点,制备载Survivin-siRNA和蛇床子阳离子脂质体纳米粒。方法:以膜材比、药脂比、超声时间和旋蒸温度为考察因素,采用正交试验法优化蛇床子素阳离子脂质体制备工艺。采用控制变量法以电位和粒径为指标,对HA-siRNA与鱼精蛋白体积比和HA-siRNA-鱼精蛋白复合物与脂质体比例进行考察。电位粒径仪测定其粒径和Zeta电位,透射电镜观察其形状;采用酶标仪测定不同浓度FAM-Survivin-siRNA标准溶液吸光度,计算载FAM-Survivin-siRNA与蛇床子素阳离子脂质体纳米粒的包封率。结果:采用薄膜分散法,通过正交试验得载蛇床子素阳离子脂质体最佳工艺为:膜材比3∶1,药脂比1∶5,旋蒸温度30℃,超声时间70 min包封率可达78.34%,通过控制变量法得载FAM-Survivin-siRNA与蛇床子素阳离子脂质体纳米粒最佳工艺为:Survivin-siRNA与鱼精蛋白体积比1∶1,鱼精蛋白复合物25μg,加入50μL阳离子脂质体。经电位粒径仪测得粒径(132.3±0.2)nm,Zeta电位(43.15±0.05)mV,其形状呈不规则的类圆形;根据标准曲线测得共载Survivin-siRNA与蛇床子素阳离子脂质体纳米粒的包封率为(81.34±0.04)%。结论:经过工艺优化制备的载Survivin-siRNA和蛇床子素阳离子脂质体纳米粒具有较好的包封率、粒径和Zeta电位。Objective:To prepare cationic liposome nanoparticles loaded with Survivin-siRNA and Cnidium monnieri based on the ability of liposomes to contain both water-soluble and lipid soluble components.Methods:The preparation technology of osthol cationic liposomes was optimized by orthogonal test with membrane material ratio,drug lipid ratio,ultrasonic time and steaming temperature as factors.The volume ratio of HA-siRNA to protamine and the ratio of HA-siRNA protamine complex to liposome were investigated by control variable method with potential and particle size as indexes.The particle size and zeta potential were measured by potentiometric particle size analyzer,and the shape was observed by transmission electron microscope;the absorbance of different concentrations of FAM-Survivin-siRNA standard solution was measured by microplate analyzer,and the entrapment efficiency of cationic liposomes loaded with FAM-Survivin-siRNA and osthole was calculated.Results:The optimum preparation conditions of osthole loaded cationic liposomes were as follows:the ratio of membrane to material was 3∶1,the ratio of drug to lipid was 1∶5,the steaming temperature was 30℃,the ultrasonic time was 70 min,and the encapsulation efficiency was 78.34%.The optimum preparation conditions of osthole loaded cationic liposomes loaded FAM-Survivin-siRNA were as follows:the volume ratio of Survivin-siRNA to protamine was 1∶1,protamine complex was 25μg.A total of 50μL cationic liposomes was added.The particle size was(132.3±0.2)nm,zeta potential was(43.15±0.05)mV,and its shape was irregular round.According to the standard curve,the entrapment efficiency of cationic liposome nanoparticles co-loaded with Survivin-siRNA and osthole was(81.34±0.04)%.Conclusion:The prepared cationic liposome nanoparticles loaded with Survivin-siRNA and osthole have good encapsulation efficiency,particle size and zeta potential.

关 键 词：SIRNA 阳离子脂质体 薄膜分散法 制备工艺 

分 类 号：R943[医药卫生—药剂学]