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

摘要

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.%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 mV，静置96 h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／mL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。