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

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。

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

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.