姜黄素纳米混悬液的抗癌活性及细胞摄取研究

In-vitro Studies on Anticancer Activity and Cellular Uptake of Curcumin Nanosuspensions

目的制备姜黄素纳米混悬液(CUR-NS)并进行体外抗癌活性及细胞摄取研究。方法采用反溶剂沉淀法制备CUR-NS。用粒度分析仪测定其粒径大小,同时用原子力显微镜进行形态学考察。采用MTT法检测姜黄素原药和CUR-NS对肿瘤细胞MCF-7的增殖抑制作用。同时,运用高效液相色谱法对药物细胞摄取进行定量研究。结果 CUR-NS的平均粒径为(69.65±0.50)nm,多分散系数为0.34±0.03,zeta电位为(-8.67±0.26)mV。原子力显微镜下观察发现,CUR-NS中粒子基本呈球形,粒径与粒度分析仪所测结果基本相符,且分散性良好,无聚合粘连。短期物理稳定性研究表明,CUR-NS在4℃下避光保存1个月后,粒径大小和分布基本保持稳定。MTT细胞毒性实验表明,24 h内姜黄素原药和CUR-NS的半数抑制浓度(IC50)分别为(44.09±0.93)μmol.L-1和(36.23±0.58)μmol.L-1,CUR-NS对肿瘤细胞MCF-7增殖的抑制作用明显强于姜黄素原药。定量摄取研究发现将姜黄素制成纳米混悬液,可在一定程度上改善其化学稳定性,增加肿瘤细胞对药物的摄取。结论纳米混悬技术是改善姜黄素类难溶性及化学不稳定药物的理化及生物活性的有效制剂方法。

Objective To develop curcumin nanosuspensions（CUR-NS）,and to study the anticancer activity and cellular uptake in vitro.Methods CUR-NS was prepared by anti-solvent precipitation method.Characterisation of the CUR-NS was investigated by dynamic laser light scattering and atomic force microscope.Cytotoxicity was evaluated by MTT assay in MCF-7 cells.High performance liquid chromatography（HPLC）was used for quantitative cellular uptake of curcumin solution and CUR-NS.Results CUR-NS with PVP as the stabiliser were successfully prepared.The mean particle size,polydispersion index and zeta potential values of CUR-NS were（69.65±0.50）nm,0.34±0.03,and（-8.67±0.26） mV,respectively.CUR-NS particles looked like sphere under atomic force microscope,and the particle diameter was in accordance with the results showed by grainsize analyzer,with good dispersibility while without polymerization or adhesion.A short-term stability study showed that CUR-NS were physically stable after storage at 4℃ for over one month.The half maximal inhibitory concentration（IC50）values of curcumin solution and CUR-NS were（44.09±0.93）and（36.23±0.58）μmol.L-1,respectively,indicating that CUR-NS were superior to curcumin solution in terms of in-vitro anticancer activity.Compared with curcumin solution,CUR-NS showed significantly higher cellular uptake.Conclusion CUR-NS can be used as a potential delivery formulation for curcumin with enhanced anticancer activity and cellular uptake.