van de Water Femke M, Boleij Johanna M, Peters Janny G P, Russel Frans G M, Masereeuw Rosalinde
Department of Pharmacology and Toxicology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
Eur J Pharm Sci. 2007 Jan;30(1):36-44. doi: 10.1016/j.ejps.2006.09.008. Epub 2006 Oct 4.
The activity of P-glycoprotein (Pgp/MDR1/ABCB1) and multidrug resistance proteins (MRP/ABCC) influence the pharmacokinetics and bioavailability of many drugs. Few suitable cell lines for the study of drug transport exist. Additional non-human cell lines may help clarify species differences and contribute to the current knowledge of drug transport. The aim of the present study was to characterize three rat epithelial cell lines for transporter expression and activity. Transporter expression was assessed in intestinal IEC-6 and renal GERP and NRK-52E cells using RT-PCR and Western blot analysis. Pgp and Mrp transport activity were analyzed by measuring calcein accumulation and glutathione-S-bimane efflux, respectively. The three cell lines showed Pgp expression and Pgp-dependent transport, both decreasing with culture time after reaching confluency. Besides Pgp, cells expressed Mrp1, Mrp3, Mrp4, and Mrp5, while Mrp2 and Mrp6 were absent. In addition, they showed temperature- and Mrp-dependent efflux of glutathione-S-bimane. Exposure to a panel of different inhibitors showed that this efflux was probably mediated by Mrp4. In conclusion, the three rat epithelial cell lines investigated showed Pgp and Mrp expression and transport. Mrp dependent transport was most likely mediated by Mrp4. In future, these cell lines may be used as in vitro models to study drug transport.
P-糖蛋白（Pgp/MDR1/ABCB1）和多药耐药蛋白（MRP/ABCC）的活性会影响许多药物的药代动力学和生物利用度。目前用于药物转运研究的合适细胞系很少。其他非人类细胞系可能有助于阐明物种差异，并为当前药物转运知识做出贡献。本研究的目的是对三种大鼠上皮细胞系的转运体表达和活性进行表征。使用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹分析对肠IEC-6细胞、肾GERP细胞和NRK-52E细胞中的转运体表达进行评估。分别通过测量钙黄绿素蓄积和谷胱甘肽-S-双硫醚外排来分析Pgp和Mrp的转运活性。这三种细胞系均显示有Pgp表达和Pgp依赖性转运，二者在达到汇合后均随培养时间而降低。除Pgp外，细胞还表达Mrp1、Mrp3、Mrp4和Mrp5，而不表达Mrp2和Mrp6。此外，它们显示出谷胱甘肽-S-双硫醚的温度依赖性和Mrp依赖性外排。暴露于一组不同的抑制剂表明，这种外排可能由Mrp4介导。总之，所研究的三种大鼠上皮细胞系均显示有Pgp和Mrp表达及转运。Mrp依赖性转运很可能由Mrp4介导。未来，这些细胞系可作为体外模型用于研究药物转运。
