叶酸介导的雷公藤甲素外泌体靶向给药系统抗恶性黑色素瘤体内外评价

In vitro and in vivo evaluation of folate-mediated triptolide exosome targeted delivery system against malignant melanoma

本研究以人恶性黑色素瘤A375细胞来源的外泌体为载体,以肿瘤细胞表面高表达的叶酸受体为靶点,以中药雷公藤甲素为模型药物,制备外泌体靶向给药系统FA-Exo/TPL,并对其体外特性和体内抑瘤效果进行表征和评价。采用梯度离心法收集外泌体,叶酸靶向修饰后包载药物,制备成的FA-Exo/TPL粒径在100 nm左右,呈茶托状双层膜结构,对药物具有较高的包封率和载药量。体外研究显示,FA-Exo/TPL可被黑色素瘤细胞大量摄取,从而增强了药物抑制细胞增殖和促进细胞凋亡的作用。体内实验结果表明,FA-Exo/TPL能有效抑制肿瘤组织的生长,延长荷瘤裸鼠的生存期,且能显著降低药物的系统毒性,起到增效减毒的作用。动物福利和实验过程均遵循复旦大学附属肿瘤医院动物伦理委员会的规定。本研究为雷公藤甲素抗恶性黑色素瘤的制剂学研究提供了新的思路和方法。

In this study,exosomes were extracted from human malignant melanoma cell A375.Folic acid(FA)receptor was used as target and triptolide(TPL)was used as model drug to prepare exosome targeted drug delivery system,FA-Exo/TPL.The physicochemical properties and antitumor effect were evaluated in vivo and in vitro.Gradient centrifugation method was applied to collect exosomes.Then,exosome was modified with FA for loading TPL.The particle sizes of the FA-Exo/TPL were about 100 nm with a double-layer membrane structure like a tray.It is characteristic of high encapsulation efficiency and drug loading.In vitro experiments showed that FA-Exo/TPL could be effectively uptaken by A375 cells,thus significantly inhibiting proliferation and promoting apoptosis the cells.In vivo experiment results showed that FA-Exo/TPL could effectively inhibit the growth of tumor tissue,prolong the model mice life cycle,and significantly reduce the systemic toxicity of the free drug,playing a synergistic and toxic role.Animal welfare and experimental procedures follow the regulations of the Animal Ethics Committee of Fudan University Shanghai Cancer Center.This study provides a new strategies and methods for the preparation of TPL against malignant melanoma.