盐酸小檗碱/叶酸-壳聚糖纳米粒的制备及其对CNE-1细胞的抑制作用  

Preparation and inhibition effects on CNE-1 cells of folate-conjugated chitosan nanoparticles loaded with berberine

作　　者：温宝芳 李文涛[1] 庄宝祥[1] 张维芬[2] 赵立民[3] 

机构地区：[1]潍坊医学院临床医学院,山东潍坊261053 [2]潍坊医学院药学院,山东潍坊261053 [3]潍坊医学院附属医院耳鼻喉头颈外科,山东潍坊261031

出　　处：《中草药》2016年第20期3594-3601,共8页Chinese Traditional and Herbal Drugs

基　　金：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)

摘　　要：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 folio acid active ester and chitosan molecules; BIUFA-CTS-NPs were prepared using ion cross-linking technique with BH as a mode/ drug. The morphology, particle size, and physicochemical characteristics such as entrapment efficiency （EE）, drug-loading, and release in vitro ofnanoparticles were studied. The effect of cell anti-migratory and anti-invasive actions of BH/FA-CTS-NPs was investigated using MTT assays, wound healingassays 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.

关 键 词：叶酸 壳聚糖 盐酸小檗碱 纳米粒 偶联 离子交联法 MTT法 划痕实验 Annexin—V-FITC单染法 抗肿瘤 抗增殖侵袭能力:凋亡 

分 类 号：R283.6[医药卫生—中药学]