木香内酯DSPE-PEG纳米胶束的制备、表征及对胶质瘤细胞凋亡的诱导作用

Preparation,characterization of micheliolide DSPE-PEG nanomicelles and its induction of apoptosis in glioma cells

目的:制备木香内酯(MCL)聚乙二醇衍生化磷脂酰乙醇胺(MCL@DSPE-PEG)纳米胶束,并对其进行表征和诱导胶质瘤细胞凋亡的研究。方法:采用薄膜水化法制备MCL@DSPE-PEG纳米胶束,观察其外观特征,检测其粒径、多分散系数(PDI)、载药量、包封率。比较MCL原料药和MCL@DSPE-PEG纳米胶束在pH 7.4枸橼酸盐缓冲液中的药物释药情况。将U87和U251胶质瘤细胞分为模型组和MCL@DSPE-PEG纳米胶束组;观察U87和U251胶质瘤细胞生长抑制情况和细胞内凋亡相关Caspase3、Caspase8、Bcl-2蛋白表达水平变化情况。结果:所制备的MCL@DSPE-PEG纳米胶束呈现大小比较均一的椭球形,其粒径为(123.8±0.5)nm,PDI为0.236±0.005,载药量为8.96%±0.56%,包封率为85.49%±3.66%,144 h的累积释放度为73.30%,而MCL原料药在48 h内已基本释放完全。MCL@DSPE-PEG纳米胶束可抑制U87和U251胶质瘤细胞生长及细胞克隆形成。与模型组比较,MCL@DSPE-PEG纳米胶束组U87和U251胶质瘤细胞的Bcl-2蛋白表达水平下降,Caspase3、Caspase8蛋白表达水平升高,差异均有统计学意义(P<0.05)。结论:成功制得MCL@DSPE-PEG纳米胶束,其具有诱导U87和U251胶质瘤细胞凋亡从而抑制肿瘤生长的作用。

Objective:To prepare and characterize micheliolide-loaded polyethylene glycol-derivatized phosphatidyle-thanolamine(MCL@DSPE-PEG)nanomicelles,and evaluate its anti-tumor effect on tumor cells.Methods:MCL@DSPE-PEG nanomicelles were prepared by film hydration method,and their appearance characteristics were observed.The particle size,polydispersion coefficient(PDI),drug-loading amount and encapsulation efficiency of the nanomicelles were detected.The drug release of MCL@DSPE-PEG nanomicelles in pH 7.4 phosphate buffer were compared within 0-144 h.The U87 and U251 glioma cells were divided into model group and MCL@DSPE-PEG nanomicelles group.Cell viability and apoptosis were evaluated.The expressions of apoptosis related protein including Caspase3,Caspase8 and Bcl-2 were detected.Results:The morphology of MCL@DSPE-PEG nanomicelles were uniform ellipsoidal shape.The particle size was(123.8±0.5)nm and PDI was 0.236±0.005.The drug-loading amount was 8.96%±0.56%,encapsulation efficiency was 85.49%±3.66%.Accumulative release rate was 73.30%within 144 h.MCL raw material was released completely within 48 h.MCL@DSPE-PEG nanomicelles inhibited the growth and cell cloning of U87 and U251 glioma cells.Compared with the model group,the expression of Bcl-2 protein was significantly decreased,while the expression of Caspase3 and Caspase8 protein were dramatically increased(P<0.05 respectively).Conclusion:MCL@DSPE-PEG nanomicelles are prepared successfully,which can induce apoptosis of U87 and U251 glioma cells and inhibit tumor growth.