青蒿琥酯纳米颗粒的制备及其体外抑制肿瘤细胞增殖的作用研究

目的：用壳聚糖（Chitosan，CS）和三聚磷酸钠（tripolyphosphate，TPP）交联制备包载青蒿琥酯纳米粒，并探讨其在体外对肿瘤细胞增殖的抑制作用。方法：以壳聚糖-三磷酸钠（CS／TPP）为基质并优化其比例，采用离子凝胶法制备包载青蒿琥酯（ART）纳米粒，对其进行表征包括粒径大小、Zeta电位、包封率、载药量和体外释放试验，以及红外光谱分析。用MTT法检测包栽青蒿琥酯的壳聚糖·三磷酸钠纳米粒对Hela、Caski、U251、MCF-7和HepG2细胞增殖的抑制作用。结果：成功构建青蒿琥酯一壳聚糖．三磷酸钠纳米颗粒（ARTnanoparticals，ART-NPs），平均粒径为166．8±0．2nnl，电位为10．2±0．79mV，红外光谱分析表明CS厂rPP成功连接并包裹ART，平均载药量和包封率分别为18％和74．82％；体外释放呈典型的双相分布，前24h呈暴发性释放（44．2％），其后缓慢释放，第9天累积释放度达67．4％。对不同肿瘤细胞的杀伤作用呈浓度和时间依赖趋势，且ART-NPs作用优于单-ART；相同浓度的ART-NPs在96h时对细胞增殖的抑制率明显高于ART组（P〈O．05）。结论：用壳聚糖和三磷酸钠交联可成功包载青蒿琥酯制成具有缓释性的纳米制剂，对肿瘤细胞的生长具有明显的抑制作用，有潜在的肿瘤治疗价值。

Preparation of Artesunate Nanoparticles and Its Inhibitory Effect on the Proferation of Cancer Cells in Vitro

Objective： To prepare the chitoson（CS）-tripolyphosphate（TPP）-Artesunate（ART） nanoparticles （ART-NPs） and inves- tigate its inhibitory effect on the proliferation of different cancer cells in vitro. Methods： Ionic gelation method was used to prepare Arte- stmate nanopartical using chitoson （CS） and tripolyphosphate （TPP） as carrier. Then the characteristics of CS/TPP nanoparticals and ART-NPs including size, zeta potential, distribution, drug-loading capacity, encapsulation efficiency and drug released were determined. Then the NPs were further characterized by FTIR. The inhibition effects of NPs on Hela, Caski, U251, MCF-7 and HepG2 cancer cells at different time （24 h, 48 h, 72 h, 96 h） were measured by MTT assay. Results： The result ofFTIR spectnun analysis showed Artesunate nanoparticles were successfully prepared. The NPs appear spherical and uniform with mean diameter of 166.8＋ 0.2 nm, its Zeta potential was 10.2＋ 0.79 mV. Their mean drug-loading amount and encapsulation efficiency were 18% and 74.82%, respectively. In vitro release test exhibited biphasic release character, that was, at the first 24 h, accumulative explosive drug release rate was 44.2% and sustained re- leased until the 9th day with accumulative release rate of 67.4%. The inhibitory effect on different cancer cells of ART-NPs was time and concentration- dependent. At 96 h, The inhibition rate of cancer cells by the NPs was higher than ones by the equate artesunate（P〈0.05）. Conclusion＂ CS/TPP nanoparticals were prepared by ionic gelation of chitosan and TPP. ART-NPs could inhibit the proliferation of dif- ferent types of cancer cells in vitro, suggesting that ART-NPs were potential candidate drug for cancer therapy.