多西紫杉醇纳米脂质体的制备及其对肝癌细胞的体内外治疗研究  

Studies on the preparation of docetaxel nano-liposomes and its treatment on liver cancer cells in vivo and in vitro

作　　者：马友龙[1] 祁海艳[1] 曹振东[1] 胡大为[1] 

机构地区：[1]承德医学院附属医院外四科,河北承德067000

出　　处：《中国生化药物杂志》2015年第6期43-47,共5页Chinese Journal of Biochemical Pharmaceutics

基　　金：河北省卫生厅科研项目(20122127)

摘　　要：目的制备多西紫杉醇(docetaxel,DOC)纳米脂质体颗粒(docetaxel nano liposome,L-DOC),研究其对肝癌细胞体内,体外的治疗作用。方法采用薄膜-超声分散法制备多西紫杉醇脂质体纳米颗粒;表征其粒度大小、Zeta电位随时间的变化情况以及包封率;采用CCK-8法检测L-DOC和DOC分别对肝癌细胞HepG2的体外抑制效果并且用流式细胞术双染法检测了细胞的死亡方式;每3天通过游标卡尺测量肿瘤体积变化,研究L-DOC和DOC分别在在体水平上对荷瘤鼠肿瘤的抑制作用;每3天称量小鼠体质量以及对小鼠主要脏器进行HE染色,研究其在体毒性。结果制备的L-DOC的平均粒度大约为104 nm,Zeta电位为-35.1 m V,静置96h后Zeta电位几乎没有变化,包封率为(71.2±1.6)%。CCK-8结果显示不同浓度的DOC分别处理细胞24 h后,均有不同程度的杀伤效果,但是相同浓度的L-DOC分别处理细胞相同时间后,抑制效果优于单独的DOC,尤其是20μg/m L的浓度,L-DOC显著比单独的DOC好(P<0.01)。此外发现L-DOC和DOC都能引起细胞凋亡。在体实验结果表明,6 mg/kg的L-DOC表现了很好的肿瘤抑制效果。6 mg/kg的L-DOC和DOC在30 d内均未对小鼠造成明显的不良反应。结论薄膜-超声分散法制备的L-DOC,粒径小,水溶性和稳定性好,在体内体外均能更好的抑制肝癌细胞,并且无不良反应。Objective To prepared the docetaxel nano liposome （L-DOC） for the therapy of liver cancer HepG2 cells in vitro and in vivo. Methods The film-ultrasonic dispersion method was used to prepare the L-DOC.The diameter and Zeta potential of L-DOC were determined by Nanosizer and the encapsulation efficiency was further measured.CCK-8 method was used to determine the cell viability of HepG2 cell after treating with various concentration of DOC and L-DOC respectively and the cell death type was detected by Flow cytometer.Next, we have studied the relative tumor volume change of tumor-bearing mice and the toxicity in vivo.Results The average diameter of L-DOC was 104 nm and the Zeta potential was about -35.1 mV.The Zeta potential of L-DOC was almost unchanged after standing for 96 hours.The encapsulation efficiency of L-DOC was （ 71.2 ±1.6 ）%.The CCK-8 results showed that the cell viability was decreased after treating with various concentration of DOC and L-DOC, but the inhibition effect of L-DOC was better than that of DOC after treating with the same dose, especially for 20μg/mL.It was found that the cell death was induced by apoptosis.The in vivo study results showed that 6mg/kg L-DOC could inhibit the tumor volume better than that of same dose of DOC.In addition, 6mg/kg L-DOC and DOC didn’ t induce in vivo toxicity.Conclusion The L-DOC is prepared by film-ultrasonic dispersion method which has small diameter, great biocompatibility.And it could inhibit the HepG2 cells in vitro and in vivo, especially for no in vivo toxicity.

关 键 词：多西紫杉醇 纳米脂质体 薄膜-超声分散法 HEPG2细胞 肿瘤治疗 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]