混合胶束递送siRNA的体外研究
In vitro study of a polymeric hybrid micelle acting as siRNA delivery vehicle
作者：唐瑶,孙逊
Author：
收稿日期： 年卷（期）页码：2020,35(06):-595-598
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：混合胶束;小干扰RNA;疫苗;树突状细胞;溶剂注入法;多分散系数;摄取能力;溶酶体逃逸能力;沉默能力;荧光定量
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基金项目：
中文摘要
目的制备混合胶束(PHM),考察其负载小干扰RNA(siRNA)的能力及所形成的复合物在树突状细胞(DC)上摄取、溶酶体逃逸及沉默的能力。方法采用溶剂注入法制备PHM,胶束亲水端带正电的聚乙烯亚胺与通过静电吸附作用复合siRNA形成载药胶束,考察载药胶束的粒径、多分散系数(PDI)和Zeta电位;通过核酸凝胶电泳确定PHM对siRNA的包载能力;考察复合物在小鼠树突状细胞系DC2.4上摄取、溶酶体逃逸和沉默的能力。结果 PHM-siRNA的平均粒径为123.7±3.4 nm,PDI为0.212±0.052,Zeta电位为13.4±2.3 mV。核酸凝胶电泳显示:氮磷比(N/P)为8:1时,PHM能完全包载siRNA。制剂在DC2.4细胞上的摄取率为50.79%±5.71%(N/P为8:1)和95.22%±1.27%(N/P为16:1),选择N/P为16:1进行后续实验。共聚焦显微镜观察结果显示:制剂可以在给药后15 h从溶酶体中逃逸出来。实时荧光定量PCR检测到PHM-siRNA对目标基因的沉默效率为66.9%±8.8%。结论 PHM-siRNA均一稳定,在DC2.4细胞上具有较高摄取、溶酶体逃逸及沉默的能力。
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