酸敏阿霉素脂质纳米粒的制备及其体外抗肿瘤活性研究

目的:本研究旨在制备具有被动靶向和酸敏特性的脂质混合纳米粒,以期提高阿霉素(doxorubicin,DOX)的靶向递药效率,降低DOX的毒副作用,提高抗肿瘤活性。方法:采用微乳法制备磷酸钙纳米粒核,薄膜分散法制备脂质混合纳米粒,硫酸铵梯度法包封DOX。采用透射电镜观察外观形态,用zeta电位及纳米粒度分析仪测定纳米粒的粒径及zeta电位,透析法评价阿霉素脂质纳米粒体外释药特征。用MTT方法研究阿霉素脂质混合纳米粒对A549细胞的细胞毒性。采用流式细胞仪和激光共聚焦显微镜观察A549细胞对阿霉素脂质纳米粒的摄取。结果:体外释药结果显示阿霉素脂质纳米粒具有酸敏特性。流式结果说明A549细胞对阿霉素脂质纳米粒的摄取具有明显的时间依赖性,激光共聚焦显示阿霉素脂质纳米粒能将阿霉素递送至细胞核中。结论:阿霉素脂质体对A549细胞有明显的细胞毒性,为进一步进行体内实验提供了基础。

The Preparation and Antitumor Activity of DOX-loaded Lipid Hybrid Nanoparticle with Acidic Sensitivity

Objective:In order to improve drug delivery efficiency, enhance the anti-tumor activity and reduce toxicity of DOX, a passive targeting lipid hybrid nanoparticle with acidic sensitivity was prepared in this study.Methods:Calcium phosphate nanoparticle was prepared by reverse micro-emulsion method, the lipid hybrid nanoparticle was prepared by film dispersion method, and the DOX was loaded by ammoniumsulfate gradients. The morphology of lipid nanoparticle was observed by transmission electron microscopy, the zeta potential and particle size were detected by Laser Particle Size Analyzer. The dialysis method was used to evaluate the characteristic of drug release of DOX-loaded lipid hybrid nanoparticle (DOX-loaded LNPS). To estimate the anti-tumor activity of DOX-loaded LNPS, the MTT method was applied for cell viability experiment in vitro. The flow cytometry and the laser confocal microscope were used to observe the uptake of the DOX-loaded LNPS in A549 cells.Results:The drug release of DOX-loaded lipid hybrid nanoparticle exhibited an acidic sensitivity property. The result of the flow cytometry indicated that the uptake of DOX-loaded LNPS in A549 cells showed time-dependent manner. The confocal microscopy also confirmed that DOX-loaded LNPS was able to deliver the DOX to the nucleus of A549 cells.Conclusion:The DOX-loaded LNPS showed significant cytotoxicity on A549 cells. It is worthy of study in vivo.