安石榴苷在MDCK细胞单层模型上的转运机制研究  

Study on the Transport Mechanism of Punicalagin in MDCK Cells Monolayer Model

作　　者：周本宏[1,2] 张红盼[1] 郭咸希[1] 李旷宇 

机构地区：[1]武汉大学人民医院,武汉430060 [2]武汉大学药学院

出　　处：《中国药师》2018年第1期1-5,共5页China Pharmacist

基　　金：国家自然科学基金项目(编号:31570349)

摘　　要：目的:采用MDCK细胞单层模型考察安石榴苷跨膜转运特性。方法:CCK8法筛选安石榴苷对MDCK细胞作用的安全浓度,Millicell-ERS测量细胞单层的TEER值确定细胞单层的完整性及致密性,考察给药浓度、方向、时间、温度、维拉帕米和EDTA-Na_2不同条件下安石榴苷的转运情况,采用HPLC法测定安石榴苷浓度,并计算表观渗透系数(P_(app))和外排率(ER)。结果:安石榴苷在MDCK模型上累积转运量具有时间和浓度依赖性,在100~300μg·ml^(-1)浓度范围内测得的顶侧(AP)到基底侧(BL)的Papp值为(6.13±0.12)×10^(-7)cm·s^(-1)、(6.96±0.26)×10^(-7)cm·s^(-1)、(5.94±0.10)×10^(-7)cm·s^(-1),未随浓度升高而增大,4℃时转运量降低,P-gp抑制药维拉帕米可以促进AP-BL方向的转运,加入EDTA-Na2后P_(app(AP-BL))显著增大。结论:安石榴苷以被动转运为主兼有主动转运参与,是P-糖蛋白(P-gp)的底物受到P-gp的外排作用,同时有细胞旁路转运途径。Objective： To investigate the transport mechanism of punicalagin MDCK monolayer model. Methods： The sate con- centration of punicalagin in MDCK cells was determined by CCK8 assay. MiLliecll - ElLS was used to measure cell monolayer TEER value to determine the integrity of the cell monolayer. The effects of direction, drug concentration, time, P-gp inhibitor and EDTA-Na2 on the absorption and transport of punicalagin were studied systematically. And then the drug concentration was analyzed by HPLC to calculate the apparent permeability coefficient （Papp） and efflux ratio（ER） . Results： Punicalagin transport in MDCK cells was time and concentration dependent. Puniealagin showed poor absorption in MDCK cells. P.ppfrom apical to basolateral side （AP-BL） within the concentration range of 100-300 μg ml-1 was （6.13±0. 12）×10-7cm·s-1 （6.96±0.26） ×10-7cm·s-1 and （5.94±0. 10） ×10-7cm·s-1 , respectively. P-gp inhibitor and EDTA-Na2 could significantly increase the transport of punicalagin in AP-BL direc- tion, while the transport decreased at 4℃. Conclusion： The transport mechanism of punicahgin might be passive diffusion as the dom- inating process involving active transportation. Punicalagin is one of P-gp substrates with exocytosis and absorbed via the paraeeUular route.

关 键 词：安石榴苷 MDCK细胞模型 跨膜转运 被动转运 

分 类 号：R945[医药卫生—微生物与生化药学]