负载连翘苷的外泌体递药系统制备及体外评价  

Preparation and in vitro evaluation of phillyrin-loaded exosomal drug delivery system

作　　者：吕田田 余海艳 薛丙权 于小玉 张燕 余亚辉 黄海英[1] LYU Tian-tian;YU Hai-yan;XUE Bing-quan;YU Xiao-yu;ZHANG Yan;YU Ya-hui;HUANG Hai-ying(Henan University of Chinese Medicine,Zhengzhou 450046,China;Agriculure and Rural Affairs of Lushi County,Sanmenxia 472100,China;Lushi County Hospital of TCM,Sanmenxia 472100,China)

机构地区：[1]河南中医药大学,郑州450046 [2]河南省卢氏县农业农村局,三门峡472100 [3]河南省卢氏县中医院,三门峡472100

出　　处：《中国新药杂志》2022年第1期89-94,共6页Chinese Journal of New Drugs

基　　金：国家自然科学基金资助项目(81904021);河南省2019年骨干教师项目(2019GGJS109);河南中医药大学研究生科研创新类项目(2020KYCX023)。

摘　　要：目的:制备负载连翘苷(phil)的外泌体(exos)递药系统(phil-exos),并考察其对人肺上皮腺癌细胞A549迁移能力的影响。方法:采用差速-超高速离心法结合超滤法获取小鼠肺泡巨噬细胞(MHS)来源的外泌体,超声法制备phil-exos并测定其粒径及电位,采用透射电子显微镜观察其形态,蛋白质免疫印迹法鉴定标志蛋白CD63和Alix,激光共聚焦显微镜观察A549细胞对phil-exos的摄取,划痕实验考察其对A549细胞迁移能力的影响。结果:phil-exos的平均粒径为(139±1)nm,聚合物分散系数(PDI)为(0.237±0.023),Zeta电位为-(19.33±0.17)mV;外观呈类球状,具有茶托状膜结构;外泌体及phil-exos对Alix高表达,CD63低表达。A549细胞8 h时对phil-exos摄取良好,且24 h时phil的迁移抑制率为46.30%,phil-exos的迁移抑制率为81.99%,phil-exos对A549细胞迁移能力具有效抑制作用。结论:phil-exos制备成功,其可显著降低A549细胞的迁移能力。Objectives:To prepare phillyrin(phil)-loaded exosomal delivery system(phil-exos) and investigate the effect on the migration of A549 cells. Methods: MHS-derived exosomes(exos) were obtained by differential ultra-high-speed centrifugation combined with ultrafiltration. phil-exos was prepared by sonicated, after which the particle size and zeta potential were measured and morphology was observed by transmission electron microscope. The marker proteins, CD63 and Alix, were identified by Western blotting. The uptake of phil-exos by A549 cells was determined using laser confocal microscopy. The effect of phil-exos on the migration of A549 cells was investigated by scratch assay. Results: The average particle size of phil-exos was(139±1) nm, PDI was(0.237±0.023) and zeta potential was(-19.33±0.17) mV. The exosomes were spherical in appearance with a distinct saucer-like membrane structure. Alix was highly expressed, and CD63 expression was low in exos and phil-exos. phil-exos can be well uptaken by A549 cells, and the inhibition of migration at 24 h was 46.30% for phil and 81.99% for phil-exos, showing phil-exos effectively inhibited the migration of A549 cells. Conclusion:The exosomal drug delivery system loaded with phillyrin was successfully prepared and the exosomes significantly inhibited the migration of A549 cells.

关 键 词：连翘苷 外泌体 小鼠肺泡巨噬细胞(MHS) 人肺上皮腺癌细胞(A549) 

分 类 号：R943[医药卫生—药剂学]