目的 制备载盐酸小檗碱（berberine hydrochloride，BH）的叶酸（folic acid，FA）偶联壳聚糖（chitosan，CTS）纳米粒（BH/FA-CTS-NPs），优化制备工艺并考察BH/FA-CTS-NPs的理化特性及其对CNE-1细胞的抑制作用。方法 利用FA活性酯与CTS分子上的氨基反应，制得FA偶联CTS（FA-CTS）；BH为模型药物，通过离子交联法制备BH/FA-CTS-NPs。考察其形态、粒径、包封率、载药量及体外释放等理化特性；分别通过MTT法、划痕实验和Annexin-V-FITC单染法进行检测，验证BH/FA-CTS-NPs对CNE-1细胞的抑制作用、抗增殖侵袭能力以及诱导凋亡作用。结果 透射电镜下观察所得BH/FA-CTS-NPs形态外观圆整，大小均匀，无粘连，平均粒径为（282.4±4.5）nm；包封率为（89.82±2.91）%；载药量为（9.16±1.01）%；5 h内累积释药率为（80.32±3.24）%，随后缓慢释放，24 h内累积释药率为（90.92±5.21）%。CNE-1细胞实验显示，BH/FA-CTS-NPs能够显著抑制CNE-1细胞的生存和增殖能力，诱导细胞CNE-1凋亡，具有剂量和时间依赖性。结论 离子交联法成功制备具有体外缓释作用的BH/FA-CTS-NPs，形态圆整，粒径大小均一，对抑制CNE-1细胞增殖及促进凋亡效果好，为开发抗肿瘤给药系统提供了理论依据。

Objective To prepare foliate-conjugated chitosan nanoparticles loaded with berberine hydrochloride (BH/FA-CTS-NPs) and investigate the optimizing technology, physicochemical characterizations, and inhibitory effect on CNE-1. Methods Folate-conjugated chitosan (FA-CTS) was prepared by amino reaction of folic acid active ester and chitosan molecules; BH/FA-CTS-NPs were prepared using ion cross-linking technique with BH as a model drug. The morphology, particle size, and physicochemical characteristics such as entrapment efficiency (EE), drug-loading, and release in vitro of nanoparticles were studied. The effect of cell anti-migratory and anti-invasive actions of BH/FA-CTS-NPs was investigated using MTT assays, wound healing assays and Annexin-V-FITC single staining assays, respectively. Results The prepared BH/FA-CTS NPs were round, and the size uniformity adhesion was not found. The results of mean particle size, EE, and drug-loading amount were (282.4±4.5) nm, (89.82±2.91)%, and (9.16±1.01)%, respectively. (80.32±3.24)% of BH in nanoparticles was released within 5 h and then released slowly, and the accumulative release rate within 24 h was (90.92±5.21)%. These results by MTT assay and wound healing assay indicated that BH/FA-CTS NPs not only inhibited the proliferation of CNE-1 cells in a concentration-and time-dependent manner, but can induced apoptosis. Conclusion BH/FA-CTS NPs with the sustained release effect could be prepared successfully by the ionic crosslinking method. Considering these properties, block proliferation and impair the migration of the CNE-1 cell line, BH/FA-CTS NPs could be an important compound for consideration in the treatment of nasopharyngeal carcinoma.

国家科技部科技计划项目（2013GA740103）；山东省自然科学基金资助项目（ZR2012CM025）；山东省科技攻关项目（2012YD18063）；山东省潍坊市科技计划项目（2014WS045）