摘要
目的:制备柚皮素壳聚糖纳米粒,初步探讨其对人肺腺癌细胞A549的细胞毒性和细胞摄取。方法:以壳聚糖和鱼精蛋白作为载体材料,采用离子胶凝法制备柚皮素壳聚糖纳米粒,透射电镜(TEM)观察其形态,马尔文激光粒度仪测定其粒径、分散度(PDI)和Zeta电位,离心法测定其包封率和载药量,采用恒温振荡水浴法对柚皮素壳聚糖纳米粒进行体外释放度研究,最后采用人肺癌细胞系A549细胞进行了细胞毒性、细胞摄取研究。结果:柚皮素壳聚糖纳米粒为球形或类球形粒子,结构完整,大小均一、球形度好,分散均匀,PDI、粒径、Zeta电位和包封率分别为0.268,139 nm、+15.7 mV和83.34%,柚皮素壳聚糖纳米粒体外释放呈缓释,24 h累积释放量达到了80%以上,体外释药过程用Higuchi方程拟合较好。MTT试验显示不同浓度的壳聚糖纳米粒和细胞作用72 h后,细胞活力均大于95%,本文所制备的壳聚糖纳米粒无细胞毒性。细胞摄取试验表明载FITC的壳聚糖纳米粒和A549细胞作用3 h后,可明显看到大量带绿色荧光的纳米粒穿过细胞膜进入细胞。结论:离子凝胶法成功制得粒径较小的柚皮素壳聚糖纳米粒,具有缓释性好,毒性小,壳聚糖纳米粒摄取率较高,可大大提高药物的利用率,具有广泛的应用前景。
OBJECTIVE To prepare naringenin chitosan nanoparticles and investigate its effect on A549 human lung adenocarcinoma cells and anti tumor activity in vitro.METHODS The naringenin chitosan nanoparticles were prepared by ionic gelation method with chitosan and protamine as carrier materials.The particle size,dispersion（PDI）,Zeta potential and morphology were investigated.The drug encapsulation efficiency of nanoparticles was determined.A549 cells of human lung adenocarcinoma were used as the model,and cytotoxicity of the nanoparticles was investigated by MTT assay.The cellular uptake of nanoparticles was investigated by fluorescence microscopy.RESULTS Nanoparticles were spherical or quasi spherical,had complete structure,uniform size with good sphericity,uniform dispersion,a PDI of 0.268,particle size of 139 nm,Zeta potential of＋15.7 mV,encapsulation efficiency of 83.34%.Naringenin chitosan nanoparticles had slow release rate in vitro,24 h cumulative release reached more than80%,which well fitted Higuchi equation.The cell viability was higher than95% and no cell toxicity was observed after different concentrations of nanoparticles acted on cells.Cellular uptake test indicated that,after FITC nanoparticles acted on A549 cells for 3 h,a large number of nanoparticles with green fluorescence entered into cells.CONCLUSION The naringenin chitosan nanoparticles prepared by ionic gelation method have small particle size,slow release rate,low toxicity,greatly improved drug uptake in tumor cells.The naringenin chitosan nanoparticles can greatly improve utilization of drugs,and have extensive application prospects.
出处
《中国医院药学杂志》
CAS
北大核心
2017年第20期2056-2060,共5页
Chinese Journal of Hospital Pharmacy
基金
辽宁省科技创新基金(编号:2014022037)
