Xenopus laevis oocytes expressing human P-glycoprotein: Probing trans- and cis-inhibitory effects on [3H]vinblastine and [3H]digoxin efflux
Xenopus laevis oocytes expressing human P-glycoprotein: Probing trans- and cis-inhibitory effects on [3H]vinblastine and [3H]digoxin efflux
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DOI：
10.1016/j.phrs.2009.07.002
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发表时间：
2010-01-01
影响因子：
9.3
通讯作者：
Endou, Hitoshi
中科院分区：
文献类型：
--
作者：
Jutabha, Promsuk;Wempe, Michael F.;Endou, Hitoshi
P-糖蛋白（P-gp; MDR 1）识别并主动转运许多结构不同的化合物（疏水性、中性和阳离子）。我们使用高通量（96孔）卵母细胞表达系统研究了MDR 1介导的药物转运。表达MDR 1的卵母细胞含有足够的ATP水平进行基本的外排研究;最佳实验温度为25 ℃。[H-3]可检测到表达MDR 1的卵母细胞的长春碱流出，得出的Km为145.5 +/- 25.4 μ M。将[H-3]长春碱（5.6 +/- 0.3 μ M）和[H-3]地高辛（1.0 +/- 0.1 μ M）分别注射到表达MDR 1的卵母细胞中，并监测其外排。奎尼丁和维拉帕米是已知的MDR 1底物/抑制剂，对MDR 1介导的[H-3]长春碱和[H-3]地高辛外排具有反式抑制作用。相反，多柔比星对MDR 1介导的[H-3]长春碱外排表现出顺式抑制而无反式抑制。表达MDR 1的卵母细胞系统为研究人员提供了一种替代的体外方法来筛选化合物，并可能允许人们探测P-gp药物-药物和/或药物-抑制剂相互作用。(C)2010爱思唯尔有限公司版权所有。
P-glycoprotein (P-gp; MDR1) recognizes and actively transports many structurally diverse compounds (hydrophobic neutral and cationic). We studied MDR1-mediated drug transport using a high-throughput (96-well) oocyte expression system. MDR1-expressing oocytes contained sufficient ATP levels to conduct fundamental efflux studies; the optimal experimental temperature was 25 degrees C. [H-3]Vinblastine efflux by MDR1-expressing oocytes was detectable and afforded a K-m of 145.5 +/- 25.4 mu M. [H-3]Vinblastine (5.6 +/- 0.3 mu M) and [H-3]digoxin (1.0 +/- 0.1 mu M) were individually injected into MDR1-expressing oocytes and their efflux monitored. Quinidine and verapamil, known MDR1 substrates/inhibitors, showed trans-inhibition on MDR1-mediated [H-3]vinblastine and [H-3]digoxin efflux. Conversely, doxorubicin demonstrated cis-inhibition without trans-inhibition on MDR1-mediated [H-3]vinblastine efflux. The MDR1-expressing oocyte system offers researchers with an alternative in vitro method to screen compounds and may allow one to probe P-gp drug-drug and/or drug-inhibitor interactions. (C) 2010 Elsevier Ltd. All rights reserved.