响应面法优化多烯紫杉醇外泌体的载药工艺  

Optimization of Docetaxel Loaded Exosomes Preparation by Response Surface Methodology

作　　者：郭密密 王婴[1] 梁达均 林定梅 王瑜桑 杨佳敏 王岩[1] GUO Mimi;WANG Ying;LIANG Dajun;LIN Dingmei;WANG Yusang;YANG Jiamin;WANG Yan(Guangdong Pharmaceutical University,Guangzhou 510006 Guangdong,China)

机构地区：[1]广东药科大学,广东广州510006

出　　处：《中药新药与临床药理》2021年第8期1189-1196,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81673608)

摘　　要：目的构建一种多烯紫杉醇的外泌体递药系统。方法采用超速离心法提取A549肿瘤细胞外泌体,电穿孔法制备多烯紫杉醇外泌体(DTX-EXO)。以包封率为考察指标,采用星点设计-响应面法考察制备工艺,动态透析法测定DTX-EXO的体外释药行为。以体外细胞摄取实验和体内小鼠活体成像实验考察外泌体的靶向性。结果DTX-EXO的最优制备工艺参数为:多烯紫杉醇浓度120μg·mL-1,电压0.75 V,电击次数5次。透射电镜下DTX-EXO呈现杯碟状,平均粒径为151 nm。DTX-EXO在体外模拟正常体液环境和肿瘤环境下24 h累计释放度分别为40.5%和56.9%。细胞摄取实验中,香豆素-6(C6)-EXO组细胞摄取荧光强度高于C6组。小鼠活体成像实验中,生理盐水组在肿瘤部位没有荧光,Cy5.5-N-羟基琥珀酰亚胺脂(Cy5.5)-DTX-EXO组在肿瘤部位的荧光最强,Cy5.5-EXO组次之,Cy5.5-DTX组最弱。结论外泌体可以用于多烯紫杉醇的传递载体,并且具有良好的缓释性和靶向性。Objective To construct an exosome delivery system of docetaxel.Methods Exosomes of A549 tumor cells were extracted by ultracentrifugation,the DTX-EXO were prepared by electroporation.The encapsulation efficiency was used as an evaluation index,the preparation process was investigated by response surface methodology,and in vitro release behavior of DTX-EXO was analyzed by dynamic dialysis.In vitro cell uptake assay and in vivo mouse imaging assay were chosen to evaluate the targeting of exosomes.Results Optimal preparation process for DTX-EXO were:concentration of DTX was 120μg·mL-1,voltage was 0.75 V,number of electric shocks was 5.Transmission electron microscope(TEM)observation showed that DTX-EXO had a typical cup-shaped structure,Nanoparticle Tracking Analysis indicated the mean diameter was 151 nm.The cumulative release of DTXEXO in 24 hours was 40.5%and 56.9%in normal environment and in tumor environment,respectively.In the cell uptake experiment,the fluorescence intensity of cells in the C6-EXO group was higher than that in the C6 group.In the in vivo imaging experiment of mouse,the normal saline group showed no fluorescence in the tumor site,while the fluorescence in the Cy5.5-DTX-EXO group was the strongest,followed by the Cy5.5-EXO group and the Cy5.5-DTX group.Conclusion Exosomes can be used to delivery DTX and have good slow-release property and targeting.

关 键 词：多烯紫杉醇 外泌体 药物传递系统 缓释性 靶向性 

分 类 号：R283.6[医药卫生—中药学]