壳聚糖纳米粒对小檗碱经Caco-2细胞单层膜转运的影响研究  

Influences of chitosan nanoparticles on the berberine transport through Caco-2 cell monolayer

作　　者：袁秀妍 徐慧慧 邵福平 尹登科 杨晔 YUAN Xiu-yan1 ,XU Hui-hui1 ,SHAO Fu-ping1 ,YIN Deng-ke1, 2,YANG Ye1, 2(1 School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012, China; 2 Institute of Pharmaceutical Preparations, Anhui Academy of Chinese Medicine, Hefei 230012, Chin)

机构地区：[1]安徽中医药大学药学院,合肥230012 [2]安徽省中医药科学院药物制剂研究所,合肥230012

出　　处：《中国新药杂志》2018年第6期708-713,共6页Chinese Journal of New Drugs

基　　金：国家自然科学基金资助项目(81303239)

摘　　要：目的:考察壳聚糖纳米粒对难吸收药物小檗碱经Caco-2细胞转运与吸收的影响。方法:采用离子凝胶法制备壳聚糖纳米粒,根据跨膜电阻和荧光素透过率验证Caco-2细胞模型的完整性,利用MTT法确定壳聚糖纳米粒和小檗碱在Caco-2细胞单层模型的合适浓度,采用高效液相色谱测定小檗碱含量,以计算小檗碱的表观渗透系数(apparent permeability coefficients,P_(app))。结果:壳聚糖纳米粒粒径为(577.07±14.89)nm,电位(24.7±0.8)mV。所建Caco-2细胞模型完整性与紧密性良好。以10μg·mL^(-1)壳聚糖纳米粒和10μg·mL^(-1)小檗碱进行细胞转运和摄取实验。纳米粒子组P_(app)(AP→BL)和P_(app)(BL→AP)分别为1.67×10^(-6)和1.48×10^(-6)cm·s^(-1);不加纳米粒子组P_(app)(AP→BL)和P_(app)(BL→AP)分别为0.55×10^(-6)和1.67×10^(-6)cm·s^(-1)。纳米粒子组AP→BL和BL→AP药物摄入量分别为(0.617 5±0.052 7)和(0.529 2±0.025 0)mg·g^(-1);不加纳米粒子组AP→BL和BL→AP药物摄入量为(0.604 7±0.061 6)和(0.406 3±0.075 6)mg·g^(-1)。结论:壳聚糖纳米粒有利于小檗碱在细胞AP→BL的转运,但不影响BL→AP的转运,这可能因为纳米粒吸附在细胞膜上,扰乱了细胞膜的稳定性,激活了膜上的离子通道或蛋白转运体,使小檗碱容易透过。Objective： To study the effect of chitosan nauoparticles on berberine transport through Caco-2 cells. Methods： Technique of ionic cross-linking was used to prepare the chitosan nanoparticles. The integrity of Caco-2 cell monolayer was investigated with the transepithelial electrical resistance of monolayer and the transmittance of fluorescein. Experiment of MTT was employed to confirm the appropriate concentration of chitosan and berberine. The concentrations of berberine in medium were analyzed by high performance liquid chromatography and the apparent permeability coefficients （Ppp） was calculated. Results： The size of chitosan nanoparticles was （577.07 ± 14.89） nm and the potential was （24.7 ±0.8 ） mV. The Caco-2 cell monolayer had good integrity and compactness. Ten microgramme per milliliter of chitosan and berberine were used for the subsequent experiment. In group with nanoparticles, the P pp and uptake of berberine from AP to BL were 1.67 ± 10^-6 cm·s^-1 and （ 0. 617 5 ± 0.0527） mg-g nanoparticles, the , and were 1.48 × 10^-6 cm-s-l and （0. 529 2 ±0. 025 0） mg·g-1 from BL to AP. In group without Papp and uptake of berberine from AP to BL were 0.55 × 10^-6 cm·s^-1 and （0. 604 7 ±0. 061 6 ）mg·g^-1 , and were 1.67×10^-6cm·s^-1 and （0.4063±0.0756） mg·g^-1 from BL to AP. Conclusion： Chitosan nanoparticles can promote the berberine transport form AP to BL through Caco-2 cell monolayer, but cannot affect the transport form BL to AP. We speculate that the absorption of nanoparticles on the cell membrane may disturb its stability, and then ion channel and protein transporter can be activated, which leads easy permeation of berberine.

关 键 词：壳聚糖纳米粒 小檗碱 CACO-2细胞 表观渗透系数 

分 类 号：R285[医药卫生—中药学] R932[医药卫生—中医学]