Department of Clinical Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev, Beer-Sheva, Israel.
Arch Gynecol Obstet. 2010 Jun;281(6):1037-44. doi: 10.1007/s00404-009-1286-7. Epub 2009 Nov 19.
To determine the role of BCRP in nitrofurantoin (NF) transport in JAr cells and the possible contribution of OATP2B1, P-gp and MRPs to this transport.
Cells were incubated with various BCRP, P-gp, MRPs, organic anion transporting polypeptide (OAT) and OATP2B1 inhibitors for 15 min, followed by incubation for 30 min with NF, with or without the inhibitors mentioned earlier. NF cytotoxicity was examined using neutral red (NR) assay. Intracellular NF levels were analyzed by HPLC.
NR assay showed that incubation conditions with NF (as carried out in our experiments) were not cytotoxic. Incubation with specific inhibitors of BCRP (FTC, Chrysin and Novobiocin), showed a significant increase in NF accumulation in the cells. Inhibitors of OATP2B1 (EGCG and BSP) had no influence on NF accumulation. Specific inhibitors of P-gp and MRPs (Verapamil and Indomethacin, respectively) also had no influence on NF accumulation in JAr cells.
NF is probably a specific substrate of BCRP, and BCRP has a major active role in NF transport in JAr cells. For the first time, we showed, that P-gp, MRPs, and the OATP2B1, probably have a negligible contribution to NF transport in JAr cells.
确定 BCRP 在 JAr 细胞中对呋喃妥因（NF）转运的作用，以及 OATP2B1、P-糖蛋白和 MRPs 对这种转运的可能贡献。
用不同的 BCRP、P-糖蛋白、MRPs、有机阴离子转运多肽（OAT）和 OATP2B1 抑制剂孵育细胞 15 分钟，然后用 NF 孵育 30 分钟，有或没有前面提到的抑制剂。用中性红（NR）测定法检测 NF 的细胞毒性。用 HPLC 分析细胞内 NF 水平。
NR 测定表明，用 NF 进行孵育的条件（如我们实验中进行的那样）没有细胞毒性。用 BCRP 的特异性抑制剂（FTC、白杨素和新生霉素）孵育，NF 在细胞内的积累显著增加。OATP2B1 的抑制剂（EGCG 和 BSP）对 NF 积累没有影响。P-糖蛋白和 MRPs 的特异性抑制剂（维拉帕米和吲哚美辛）对 JAr 细胞中 NF 积累也没有影响。
NF 可能是 BCRP 的特异性底物，BCRP 在 JAr 细胞中对 NF 的转运中起主要的主动作用。我们首次表明，P-糖蛋白、MRPs 和 OATP2B1 可能对 JAr 细胞中 NF 的转运贡献微不足道。
