疏水化阳离子透明质酸胶束介导的siRNA转染影响因素及细胞靶向性研究  

Influencing factors of hydrophobic modified hyaluronic acid-based cationic micelles and cell targeting in siRNA transfection

作　　者：沈雁[1] 李谦[2] 范达[1] 李瑞珺[1] 涂家生[1] 

机构地区：[1]中国药科大学药剂学教研室,南京210009 [2]中国药科大学生命科学与技术学院,南京210009

出　　处：《中国药科大学学报》2012年第2期124-129,共6页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.81072588);中央高校基本科研业务费专项资金资助项目(No.JKQ2011018)~~

摘　　要：研究疏水化阳离子透明质酸胶束HAPL9600介导基因转染的影响因素。通过测定HAPL9600胶束与siRNA形成复合物的粒径,分别考察透明质酸相对分子质量、HAPL9600浓度、HAPL9600与siRNA比例、pH、介质、孵育时间以及血清对疏水化阳离子透明质酸胶束体外转染效率的影响,筛选较优的转染条件。同时进行受体抑制实验,考察该载体的细胞靶向性。结果表明当透明质酸相对分子质量为9 600、载体与siRNA质量比为8,孵育时间为30 min、体系中HAPL9600浓度为15 mg/mL,转染介质为pH 6.8的PBS溶液时转染效率较高。游离透明质酸可抑制经CD44介导的siRNA的转染效率。HAPL9600可作为合成新型靶向非病毒载体的骨架,但必须控制有关因素才能得到可重复的最佳转染结果。The objective was to investigate the influencing factors on the siRNA delivery efficiency of hydrophobic modified hyaluronic acid with molecular weight 9 600 grafted with spermine and laurylamine cationic micelles（HAPL9600）.The transfection efficiency of HAPL9600/siRNA was determined by flow cytometry.The influence of the molecular weight of HA,complexes forming time,the concentration of the HAPL9600,pH,the transfection medium and the weight ratio of HAPL9600 to siRNA on transfection were investigated.The competitive inhibition experiment was studied to investigate the in vivo cell target ability.The results indicated that HAPL9600 would reach the optimal transfection efficiency when the molecular weight of HA is 9 600 with 30 min of complexes forming time,15 mg/mL of concentration of the HAPL9600,8 of the weight ratio of HAPL9600 to siRNA at PBS（pH 6.8） transfection medium.The transfection efficiency of the HAPL9600 was reduced by competitive binding of free HA onto the CD44 receptors,thereby lowering the extent of cellular uptake via CD44 receptor mediated endocytosis,which suggested its in vitro targeting ability on the tumor cells.In all,HAPL9600 could obtain optimal,reduplicate and targeting transfection results by controlling related factors.

关 键 词：SIRNA 疏水改性 透明质酸胶束 细胞靶向 转染效率 

分 类 号：R96[医药卫生—药理学]