硬脂醇半乳糖苷修饰阿西替尼脂质体的制备及体外细胞毒性研究  

Preparation and in vitro cytotoxicity of stearyl alcohol galactosidase-modified axitinib liposomes

作　　者：方晓旭 郭伟英 FANG Xiao-xu GUO Wei-ying(College of Pharmacy, Jinzhou Medical University, Jinzhou 121 O01, China)

机构地区：[1]锦州医科大学药学院,锦州121001

出　　处：《中国新药杂志》2017年第3期343-350,共8页Chinese Journal of New Drugs

基　　金：辽宁省科技厅计划项目(2011402015)

摘　　要：目的:研究硬脂醇半乳糖苷修饰的阿西替尼脂质体对人肝癌SMMC-7721细胞株的增殖及凋亡的诱导作用。方法:采用冷冻干燥法制备硬脂醇半乳糖苷修饰的阿西替尼脂质体,以包封率为评价指标,采用葡聚糖凝胶过滤法和高效液相色谱法测定硬脂醇半乳糖苷修饰的阿西替尼脂质体的包封率。采用Box-Behnken响应面设计法优化制备工艺,并研究CCK-8法检测硬脂醇半乳糖苷修饰的阿西替尼脂质体对人肝癌SMMC-7721细胞株的抑制情况,采用Annexin V/PI流式细胞分析法检测细胞凋亡,Western blot检测凋亡蛋白Bax,Bcl-2,P53的表达。结果:最佳工艺:药脂比为1∶20,胆脂比为1∶8,硬脂醇半乳糖苷与磷脂比为1∶15,水合温度为55℃。CCK-8法检测不同浓度的硬脂醇半乳糖苷修饰的阿西替尼脂质体对SMMC-7721细胞株的增殖具有抑制作用,呈现时间和浓度依赖性。Annexin V/PI流式细胞分析法检测硬脂醇半乳糖苷修饰的阿西替尼脂质体对人肝癌SMMC-7721细胞株的抑制率优于人肺癌A549细胞株,Western blot检测发现随着硬脂醇半乳糖苷修饰的阿西替尼脂质体浓度的升高,Bax,P53蛋白的表达逐渐升高,Bcl-2蛋白的表达逐渐降低。结论:硬脂醇半乳糖苷修饰的阿西替尼脂质体在一定浓度范围内,可在体外抑制人肝癌SMMC-7721细胞株的增殖并诱导其凋亡,且随着硬脂醇半乳糖苷修饰的阿西替尼前体脂质体的浓度升高,凋亡率显著增加,可能的机制是增强促凋亡蛋白Bax,P53的表达,降低抗凋亡蛋白Bal-2的表达。硬脂醇半乳糖苷修饰的阿西替尼脂质体诱导SMMC-7721细胞株凋亡能力比A549强,初步判定硬脂醇半乳糖苷修饰的阿西替尼脂质体具有主动肝靶向性。Objective：To study the effect of stearyl alcohol galactosidase-modified axitinib liposomes in inducing proliferation and apoptosis of liver cancer SMMC-7721 cells. Methods：Stearyl alcohol galactosidase modified-axitinib liposomes were prepared by freeze-drying. The entrapment efficiency was measured by sephadex filtration and high performance liquid chromatography. The liposomes were optimized by Box-Behnken response surface design experiment. The inhibitory effect of stearyl alcohol galactosidase-modified axitinib liposomes on growth of SMMC-7721 cells was tested by CCK-8 assay. The apoptosis of SMMC-7721 cells and A549 cells was assayed by Annexin V/PI flow cytometry. The expressions of Bax,P53 and Bcl-2 in SMMC-7721 cells were detected by western blot. Results：The best prescription was as follows：axitinib to lecithin ratio was 1 ∶ 20,cholesterol to lecithin ratio was 1∶ 8,stearyl alcohol galactosidase to lecithin ratio was 1∶ 15,the optimum temperature was 55 ℃.The stearyl alcohol galactosidase-modified axitinib liposomes inhibited the growth of SMMC-7721 cells in a timeand concentration-dependent manner. The inhibition effect of stearyl alcohsidase-modified axitinib liposomes onSMMC-7721 cells were better than A549 cells under the same conditions. Western blot displayed that the expression levels of Bax and P53 proteins increased while that of Bcl-2 protein decreased with increasing concentration.Conclusion：The stearyl alcohol galactosidase-modified axitinib liposomes can inhibit the proliferation and induce the apoptosis of SMMC-7721 cells in vitro by up-regulating the expressions of Bax and P53 and down-regulating the expression of Bcl-2. The apoptosis rate of SMMC-7721 cells increases with the increasing concentration of stearyl alcohol galactosidase-modified axitinib liposomes. Stearyl alcohol galactosidase-modified axitinib liposomes have stronger apoptosis-inducing effect for SMMC-7721 cells than A549 cells. It is preliminarily concluded that stearyl alcohol galactosidase-modified axit

关 键 词：阿西替尼 脂质体 硬脂醇半乳糖苷 细胞毒性 

分 类 号：R979.1[医药卫生—药品]