摘要
目的研究制备磁性吉西他滨隐形纳米脂质体（MGSL）的最佳条件并考察其理化性质以及在体外的抑瘤效应。方法通过逆相蒸发法制备MGSL，采用扫描电镜和原子力显微镜对其形态进行观察；利用激光粒度分析仪测定MGSL粒径大小和粒度分布；通过高效液相色谱法（HPLC）检测药物的载药量和包封率；使用专业磁性测试仪进行体外磁响应性测定。观察MGSL诱导的乳腺癌MCF-7细胞凋亡过程中的细胞形态学改变，MTT法获得MGSL作用乳腺癌细胞生长抑制率，流式细胞术分析MGSL引起的细胞凋亡率和细胞周期分布。结果MGSL为圆形或椭圆形，大小均匀-致，其平均粒径为206．6纳米，粒度分布窄，大小均匀。MGSL载药量为（10．4±0．7）％，包封率为（81．7±5．1）％。在体外，MTT实验结果与MGSL及游离的吉西他滨相对照，MGSL对人乳腺癌细胞有明显的杀伤作用，而且该作用随浓度升高有上升趋势；作用2h时在相同的浓度下较裸药组的抑瘤作用低（P〈O．05），但作用48h后两组的抑制率无差别（P〈0．05）；另外MGSL比磁性吉西他滨纳米脂质体（MGL）的抑制率略低，但无统计学差异（P〉0．05）；乳腺癌细胞周期分布结果显示，MGSL是主要作用于s期的细胞周期药物；另外流式细胞术也证实了MGSL还有促乳腺癌细胞凋亡的作用，其诱导凋亡率达到51．62％。结论本法制备的MGSL符合作为纳米磁靶向给药系统的条件，其在体外显示了较好的抑瘤效应，可望成为一种有效的抗肿瘤物质。
Objective To explore a new approach to prepare magnetic gemcitabine stealth nano-liposomes（MGSL）, and evaluate its quality and its antitumor effects in vitro. Methods MGSL was prepared by reverse-phase evaporation method. The coated rate, shape, diameter, distribution of MGSL were examined by high performance liquid chromatography （HPLC）, microscopy and laser light scattering. Then the stability of MGSLs was also evaluated. The antitumor effect on breast cancer cell line MCF-7 in vitro was investigated. Electron microscopy was employed to observe morphological features and uhrastrueture changes of apoptosis of MCF-7 cells. The effect of MGSL on proliferation inhibitory rate of MCF-7 cells was measured with MTr method. The FCM analysis was carried out to examine the cell cycle distribution and cell apoptotie rate. Results The electron microscope and atomic force microscope photographs of MGSL showed that the shape of liposome was round or elliptical. It was showed that 97.62 percent of the liposome optimized was 206.6nm with above the coated rate 81.7 % . Moreover, it possessed a good magnetic response and a high stability. MCF-7 cell line was sensitive to MGSL and the eytotoxity was correlated with the loaded drug dose. After treated for 2h, the effect of MGSL on proliferation inhibitory rate of MCF-7 cells was lower than that of gemeitabine group（ P 〈 0.05）, while treated for 48h, the effect on proliferation inhibitory rate was similar（ P 〉 0.05） . Moreover, the effect on proliferation was also similar between MGSL group and MGL group（ P 〉 0.05）. The effect of apoptosis of MGSL on MCF-7 was obvious and the rate of apoptosis was 51.62% . Conclusion MGSL prepared by reverse-phase evaporation method could meet with the demand of targeted delivery system, and it might be an effective anti-tumor agent.
出处
《解剖学报》
CAS
CSCD
北大核心
2009年第1期7-12,共6页
Acta Anatomica Sinica
基金
“十五”期间国家高技术研究发展计划(863计划)(2001AA218051)资助
