噻吩诺啡在Caco-2细胞上的转运特征  

Transport of thiophenorphine across Caco-2 monolayer model

作　　者：李峥[1] 庄笑梅[1] 李素云[2] 张振清[1] 阮金秀[1] 

机构地区：[1]军事医学科学院毒物药物研究所药物代谢与药代动力学重点实验室,北京100850 [2]北京世纪坛医院,北京100038

出　　处：《中国药理学与毒理学杂志》2010年第1期64-68,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：国家重大新药创制科技重大专项资助项目(2009ZX09301-002);国家重大新药创制科技重大专项资助项目(2009ZX09304-004)~~

摘　　要：目的探讨噻吩诺啡在Caco-2细胞上的转运特征及其对P-糖蛋白(P-gp)功能和表达的影响。方法采用高效液相色谱-质谱-质谱法测定噻吩诺啡浓度,研究噻吩诺啡在单层细胞中的双向转运,考察时间及转运蛋白抑制剂对噻吩诺啡在Caco-2细胞上转运的影响;流式细胞仪检测胞内钙黄绿素-AM浓度,评价噻吩诺啡对P-gp的抑制作用;采用Western蛋白印迹法检测P-gp表达。结果噻吩诺啡通过Caco-2单层细胞的转运量在1.5h内随时间延长呈线性增加,表观渗透系数(Papp)2.338×10-6cm.s-1;加入P-gp及多药耐药相关蛋白2(MRP2)抑制剂环孢素A和MK571后分别提高2.8和2.3倍;在噻吩诺啡作用下,胞内钙黄绿素-AM浓度无显著变化,P-gp表达无显著增加。结论噻吩诺啡在Caco-2细胞吸收中等偏差,是P-gp和MRP2的共同底物,噻吩诺啡对P-gp无诱导或抑制作用。OBJECTIVE To explore the absorption mechanism of thiophenorphine,and its effect on P-glycoprotein（P-gp） expression by using Caco-2 cell monolayer model.METHODSThe LC-MS-MS method was applied to determine thiophenorphine concentration in millicell system.The bi-directional permeability studies were performed to investigate the potential involvement of efflux carriers in the intestinal absorption.P-gp inhibition was studied by flow cytometry using calcein-AM as P-gp substrate.The expression of P-gp was evaluated using Western blotting.RESULTSThiophenorphine transport in Caco-2 cells was in time-dependent manner.Its average apparent permeability coefficient（Papp） was 2.338×10^-6 cm·s^-1.Papp was increased 2.8 folds by P-gp inhibitor ciclosporin A,and 2.3 folds by mulitdrug resistance-associated protein2（MRP2） inhibitor MK571.The accumulation of calcein-AM and the expression of P-gp in Caco-2 cell line wasn＇t changed noticeably by thiophenorphine.CONCLUSION Thiophenorphine is a common substrate of P-gp and MRP2 and it shows normal transport in millicell system.The expression of P-gp doesn＇t induce by thiophenorphine.

关 键 词：噻吩诺啡 色谱法 高压液相 细胞 培养的 糖蛋白类 多药耐药相关蛋白类 

分 类 号：R969.1[医药卫生—药理学]