线粒体靶向的载紫杉醇TPP-PEG-PE纳米胶束的体外评价及促肿瘤细胞凋亡研究

目的制备载紫杉醇TPP-PEG-PE纳米胶束,研究其体外释放行为,考察其线粒体靶向性及促A549肺癌细胞凋亡效果。方法采用薄膜水化法制备载紫杉醇TPP-PEG-PE纳米胶束,以载药量、包封率、粒径为考察因素,优选载紫杉醇TPP-PEG-PE纳米胶束的制备工艺参数,再对优选工艺的纳米载药系统进行表征,采用体外释药、线粒体靶向、肺癌细胞毒性和细胞凋亡实验对该载药系统进行评价。结果载紫杉醇TPP-PEG-PE纳米胶束粒径为(18.7±0.8)nm,Zeta电位为(13.4±0.5)m V,透射电子显微镜结果表明,载紫杉醇TPP-PEG-PE纳米胶束为大小较为均一的规则圆球型。线粒体靶向实验表明TPP-PEG-PE纳米胶束可以促进药物聚集在线粒体部位,肺癌细胞毒性实验显示载紫杉醇TPP-PEG-PE纳米胶束抗细胞凋亡效果良好,Hoechst染色提示凋亡肺癌细胞出现了大量形态学改变,载紫杉醇TPP-PEG-PE纳米胶束可以明显提高促凋亡Caspase-3活性以及减少抗凋亡蛋白Bcl-2和c-IAP1表达量,均显著优于载紫杉醇PEG-PE纳米胶束和紫杉醇组的实验结果(P0.01)。结论载紫杉醇TPP-PEG-PE纳米胶束具有良好的肺癌细胞线粒体靶向性和促肺癌细胞凋亡作用,是一种潜在高效的肺癌细胞线粒体靶向给药系统。

In vitro evaluation of mitochondrial-targeted taxol TPP-PEG-PE nanomicelles and promotion of tumor cell apoptosis

Objective To prepare taxol TPP-PEG-PE nanomicelles and study its in vitro release behavior, mitochondrial targeting and pro-apoptosis of A549 lung cancer cells. Methods The taxol TPP-PEG-PE nanomicelles were prepared by membrane hydration method. Based on the drug loading, encapsulation efficiency and particle size, the preparation parameters of taxol TPP-PEG-PE nanomicelles were optimized. The preferred nano-drug delivery system was then characterized. The drug delivery system was evaluated by in vitro drug release, mitochondrial targeting, lung cancer cell toxicity, and apoptosis assay. Results The diameter of taxol TPP-PEG-PE nanomicelles was (18.7 ±0.8) nm, Zeta potential was (13.4 ±0.5) mV, and the results of TEM electron microscopy showed that the taxol TPP-PEG-PE nanomicelles were regular spheres of uniform size. Mitochondrial targeting experiments showed that TPP-PEG-PE nanomicelles can promote drug accumulation in mitochondrial sites. Lung cancer cytotoxicity assay showed that taxol TPP-PEG-PE nanomicelles had good anti-apoptotic effect, and Hoechst staining suggested that a large number of morphological changes were observated in apoptotic lung cancer cells. Taxol TPP-PEG-PE nanomicelles could significantly increase the pro-apoptotic Caspase-3 activity and reduce the expression of anti-apoptotic protein Bcl-2 and c-IAP1. They were all significantly superior to that of taxol-PEG-PE nanomicelles and taxol group (P < 0.01). Conclusion The taxol TPP-PEG-PE nanomicelles had good mitochondrial targeting of lung cancer cells and promoted the apoptosis of lung cancer cells. It was a potential and efficient drug delivery system for lung cancer cell mitochondria.