Multiple Efflux Pumps Are Involved in the Transepithelial Transport of Colchicine: Combined Effect of P-Glycoprotein and Multidrug Resistance-Associated Protein 2 Leads to Decreased Intestinal Absorption Throughout the Entire Small Intestine
Multiple Efflux Pumps Are Involved in the Transepithelial Transport of Colchicine: Combined Effect of P-Glycoprotein and Multidrug Resistance-Associated Protein 2 Leads to Decreased Intestinal Absorption Throughout the Entire Small Intestine
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DOI：
10.1124/dmd.109.028282
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发表时间：
2009-10-01
影响因子：
3.9
通讯作者：
Amidon, Gordon L.
中科院分区：
文献类型：
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作者：
Dahan, Arik;Sabit, Hairat;Amidon, Gordon L.
本研究的目的是彻底表征参与口服微管聚合抑制剂秋水仙碱的肠渗透性的外排转运蛋白，并评估这些转运蛋白在限制其口服吸收中的作用。研究了P-糖蛋白（P-gp）、多药耐药相关蛋白2（MRP 2）和乳腺癌耐药蛋白（BCRP）抑制剂对跨Caco-2细胞单层的秋水仙碱双向渗透性的影响，同时抑制一种与多种转运蛋白。秋水仙碱的渗透性，然后在大鼠小肠的不同区域进行了研究，通过在原位单程灌注。免疫印迹法检测不同肠段P-gp/MRP 2表达水平的相关性。P-gp抑制剂[N-（4-[2-（1，2，3，4-四氢-6，7-二甲氧基-2-异喹啉基）乙基]-苯基）-9，10-二氢-5-甲氧基-9-氧代-4-吖啶甲酰胺（GF 120918）、维拉帕米和奎尼丁]和MRP 2抑制剂[3-[[3-[2-[3-[3-[2-[3-[3-[2-[3-[3-[2-[3-[3-[2-[3-[3-[2-[3-[3-[2-[3-[3-[2-[3-[3-[2-[3-[3 - 3-[3-[3 - 3-[3 - 3 - 3（7-氯喹啉-2-基）乙烯基]苯基]（2-二甲基氨基甲酰基乙基硫烷基）甲基硫烷基]丙酸（MK 571），吲哚美辛，和对氨基马尿酸（p-AH）]以浓度依赖性方式显著增加顶侧（AP）-基底侧（BL）转运和降低BL-AP Caco-2转运。BCRP抑制剂烟曲霉素C（FTC）和泮托拉唑无影响。P-gp/MRP 2抑制剂组合大大减少了秋水仙碱粘膜分泌，包括完全消除外排（GF 120918/MK 571）。秋水仙碱显示低（与美托洛尔）和恒定的渗透性沿着大鼠小肠。GF 120918显著增加了回肠中秋水仙素的渗透性，而在空肠中没有影响，而MK 571增加了空肠的渗透性，而不改变回肠转运。GF 120918/MK 571组合在空肠中引起与单独MK 571相似的作用，在回肠中引起与单独GF 120918相似的作用。P-gp表达从近端到远端呈梯度增加，而MRP 2从近端到远端小肠区呈下降趋势。总体而言，研究表明，P-gp和MRP 2（而非BCRP）的联合作用主导了秋水仙碱跨上皮转运，导致完全覆盖整个小肠，并使外排转运主导了肠道渗透性过程。
The purpose of this study was to thoroughly characterize the efflux transporters involved in the intestinal permeability of the oral microtubule polymerization inhibitor colchicine and to evaluate the role of these transporters in limiting its oral absorption. The effects of P-glycoprotein (P-gp), multidrug resistance-associated protein 2 (MRP2), and breast cancer resistance protein (BCRP) inhibitors on colchicine bidirectional permeability were studied across Caco-2 cell monolayers, inhibiting one versus multiple transporters simultaneously. Colchicine permeability was then investigated in different regions of the rat small intestine by in situ single-pass perfusion. Correlation with the P-gp/MRP2 expression level throughout different intestinal segments was investigated by immunoblotting. P-gp inhibitors [N-(4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2-isoquinolinyl)ethyl]-phenyl)-9,10-dihydro-5-methoxy-9-oxo-4-acridine carboxamide (GF120918), verapamil, and quinidine], and MRP2 inhibitors [3-[[3-[2-(7-chloroquinolin-2-yl)vinyl]phenyl](2-dimethylcarbamoylethylsulfanyl)methylsulfanyl] propionic acid (MK571), indomethacin, and p-aminohippuric acid (p-AH)] significantly increased apical (AP)-basolateral (BL) and decreased BL-AP Caco-2 transport in a concentration-dependent manner. No effect was obtained by the BCRP inhibitors fumitremorgin C (FTC) and pantoprazole. P-gp/MRP2 inhibitors combinations greatly reduced colchicine mucosal secretion, including complete abolishment of efflux (GF120918/MK571). Colchicine displayed low (versus metoprolol) and constant permeability along the rat small-intestine. GF120918 significantly increased colchicine permeability in the ileum with no effect in the jejunum, whereas MK571 augmented jejunal permeability without changing the ileal transport. The GF120918/MK571 combination caused an effect similar to that of MK571 alone in the jejunum and to that of GF120918 alone in the ileum. P-gp expression followed a gradient increasing from proximal to distal segments, whereas MRP2 decreased from proximal to distal small intestinal regions. Overall, it was revealed that the combined effect of P-gp and MRP2, but not BCRP, dominates colchicine transepithelial transport, leading to complete coverage of the entire small intestine, and makes the efflux transport dominate the intestinal permeability process.