共载盐酸阿霉素和水飞蓟宾口服肝靶向脂质体机制评价

Targeting mechanism evaluation of oral hepatic targeting liposomes co-encapsulated doxorubicin hydrochloride and silybin

目的:研究基于胆酸转运体的DSPE-PEG-胆酸修饰脂质体体外细胞转运和摄取及其机制。方法:构建共载盐酸阿霉素(DOX)和水飞蓟宾(silybin)的DSPE-PEG-胆酸修饰脂质体(CA-LP-DOX/silybin),采用表面等离子体共振(surface plasmon resonance,SPR)技术考察CA-LP-DOX/silybin与细胞的相互作用;采用Caco-2细胞转运考察CA-LP-DOX/silybin经肠吸收机制;采用HepG2细胞摄取考察CA-LP-DOX/silybin经肝细胞摄取机制。结果:所建立的CA-LP-DOX/silybin与Caco-2细胞和HepG2细胞均可相互作用,且与HepG2细胞相互作用更强。与未修饰脂质体(LP-DOX/silybin)和游离药物(DOX/silybin)相比,其经Caco-2细胞转运和Hep G2细胞摄取均增强,其摄取量受温度、浓度和内吞抑制剂影响。结论:CA-LP-DOX/silybin能够增强DOX和silybin的细胞转运和摄取,其机制为胆酸转运体介导,且有网格蛋白和小窝蛋白参与的内吞过程。

Objective:To study the in vitro cell transport,uptake by HepG2 cells and uptake mechanism of DSPE-PEG-cholic acid modified liposome co-encapsulated silybin and doxorubicin hydrochloride(DOX).Methods:Preparing the DSPE-PEG2000-cholic acid modified liposomes co-encapsulated silybin and doxorubicin hydrochloride(CA-LP-DOX/silybin).Using surface plasmon resonance(SPR)technology to determine the interaction of CALP-DOX/silybin with HepG2 cells and Caco-2 cells.To study the gastrointestinal absorption mechanism of CA-LPDOX/silybin by in vitro Caco-2 cells transport experiment.To study hepatic uptake characteristics and mechanism of CA-LP-DOX/silybin by in vitro HepG2 cells uptake experiment.Results:The CA-LP-DOX/silybin built has good interaction with Caco-2 cells and HepG2 cells,and the interaction with HepG2 cells is stronger than Caco-2 cells.Compared with the unmodified liposomes(LP-DOX/silybin)and raw materials(DOX/silybin),the CA-LP-DOX/silybin’s transport and uptake are both decreased across Caco-2 cells monolayer and by HepG2 cells in vitro.What’s more,the uptake is affected by temperature,drug concentration and endocytosis inhibitors.Conclusion:CA-LPDOX/silybin can improve the transport and uptake of DOX and silybin,and the mechanism involves cholic acid receptor mediated reaction and caveolin and clathrin protein mediated endocytosis pathway.