线粒体靶向性的姜黄素TPP-PEG-PLGA脂质体的制备及促肝癌细胞凋亡研究

目的 研究姜黄素3-苯羧基-三苯基溴化膦-聚乙二醇-聚乳酸聚乙醇酸共聚物(TPP-PEG-PLGA)脂质体的线粒体靶向性及促肝癌细胞凋亡效果。方法 以薄膜水化法制备姜黄素TPP-PEG-PLGA脂质体,评价该载药系统的稳定性、溶血、体外释放及体内代谢情况;以荧光实验研究脂质体的肝癌细胞摄取、线粒体靶向和肝肿瘤组织靶向效果;在等剂量给药条件下,评价姜黄素TPP-PEG-PLGA脂质体促肝癌细胞凋亡效果。结果 姜黄素TPP-PEG-PLGA脂质体呈圆球型结构,具有较好的稳定性、较小的溶血率和良好的缓释药物性能;药动学结果显示,姜黄素TPP-PEG-PLGA脂质体能将药物的半衰期延长3倍,生物利用度提高4倍;荧光实验显示,TPP阳离子能促进脂质体的细胞摄取,靶向进入线粒体,还可以提高药物在肝肿瘤组织部位的靶向和滞留效果;细胞药效显示,姜黄素TPP-PEG-PLGA脂质体能明显升高肿瘤细胞内H₂O₂水平,降低线粒体膜电位,提高促凋亡蛋白Bax表达,明显降低抗凋亡蛋白BCl-2表达,这些细胞凋亡试验结果均明显优于姜黄素脂质体和姜黄素。结论 本研究表明姜黄素TPP-PEG-PLGA脂质体具有良好的线粒体靶向功能,能增强药物促肝癌细胞凋亡效果。

Preparation of Mitochondrial Targeting Curcumin TPP-PEG-PLGA Iposomes and Study on Promoting Hepatoma Cell Apoptosis

OBJECTIVE To study the mitochondrial targeting of curcumin TPP-PEG-PLGA liposomes and its effect on promotingapoptosis of hepatocellular carcinoma cells.METHODS Curcumin TPP-PEG-PLGA liposomes were prepared by the thin film hydration method. The stability, hemolysis, in vitro release and in vivo metabolism of the system were evaluated. Fluorescence assay was used to study the effects of liposomes uptake in hepatoma cells, mitochondrial targeting and liver tumor tissue targeting. Under the conditions of equal dose administration, the effect of curcumin TPP-PEG-PLGA liposomes on promoting apoptosis of liver cancer cells was evaluated.RESULTS Curcumin TPP-PEG-PLGA liposomeswere in regular round spheres, the liposomes had good stability, low hemolysis rate and good sustained release properties. Pharmacokinetic results showed that curcumin TPP-PEG-PLGA liposomes could prolong the half-life of the drug by 3 times and increase the bioavailability by 4 times. Fluorescence test showed that TPP cations could promote the uptake of liposomes into mitochondria, and also improve the targeting and retention effect of drugs in liver tumor tissue. Cell efficacy showed that curcumin TPP-PEG-PLGA liposomes could significantly increase H₂O₂ level, decrease mitochondrial membrane potential, increase the expression of pro-apoptotic protein Bax, and significantly reduce the expression of anti-apoptotic protein Bcl-2 in tumor cells. The results of curcumin TPP-PEG-PLGA liposomes on promoting apoptosis of liver tumor cells were significantly better than that of curcumin liposomes and curcumin.CONCLUSION This study showed that curcumin TPP-PEG-PLGA liposomes had a good mitochondrial targeting function and can enhance the drug promoting apoptosis of liver tumor cells.