Polarized location of SLC and ABC drug transporters in monolayer-cultured human hepatocytes
Polarized location of SLC and ABC drug transporters in monolayer-cultured human hepatocytes
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DOI：
10.1016/j.tiv.2015.03.019
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发表时间：
2015-08-01
影响因子：
3.2
通讯作者：
Fardel, Olivier
中科院分区：
文献类型：
--
作者：
Le Vee, Marc;Jouan, Elodie;Fardel, Olivier
以单层结构培养的人肝细胞代表了肝脏毒理学中成熟的体外模型，尤其用于药物转运蛋白研究。药物转运蛋白的极化状态，即，然而，在这些培养的肝细胞中，它们在窦状隙或小管膜上的协调位置仍然没有完全记录。因此，本研究的目的是分析转运蛋白的表达和定位在这样的细胞。大多数药物转运蛋白首先在单层培养的人肝细胞中以显著的mRNA水平存在。培养的人肝细胞形态学上表现出胆小管样结构，随后通过免疫荧光染色证明，在其正弦极表达内流转运蛋白有机阴离子转运多肽（OATP）1B1、OATP 2B1和有机阳离子转运蛋白（OCT）1以及外排转运蛋白多药耐药相关蛋白（MRP）3。此外，在单层培养的人肝细胞的小管极检测到外排转运蛋白P-糖蛋白和MRP 2。此外，观察到外排转运蛋白胆盐输出泵、MRP 2和P-糖蛋白的参考底物的小管分泌以及窦状隙药物转运蛋白活性。药物转运蛋白在单层培养的人肝细胞中的这种极化和功能性表达突出了在异生物质转运研究中使用这种人体外细胞模型的兴趣。(C)2015爱思唯尔有限公司版权所有。
Human hepatocytes cultured in a monolayer configuration represent a well-established in vitro model in liver toxicology, notably used in drug transporter studies. Polarized status of drug transporters, i.e., their coordinated location at sinusoidal or canalicular membranes, remains however incompletely documented in these cultured hepatocytes. The present study was therefore designed to analyze transporter expression and location in such cells. Most of drug transporters were first shown to be present at notable mRNA levels in monolayer-cultured human hepatocytes. Cultured human hepatocytes, which morphologically exhibited bile canaliculi-like structures, were next demonstrated, through immunofluorescence staining, to express the influx transporters organic anion transporting polypeptide (OATP) 1B1, OATP2B1 and organic cation transporter (OCT) 1 and the efflux transporter multidrug resistance-associated protein (MRP) 3 at their sinusoidal pole. In addition, the efflux transporters P-glycoprotein and MRP2 were detected at the canalicular pole of monolayer-cultured human hepatocytes. Moreover, canalicular secretion of reference substrates for the efflux transporters bile salt export pump, MRP2 and P-glycoprotein as well as sinusoidal drug transporter activities were observed. This polarized and functional expression of drug transporters in monolayer-cultured human hepatocytes highlights the interest of using this human in vitro cell model in xenobiotic transport studies. (C) 2015 Elsevier Ltd. All rights reserved.