Menez Cécile, Buyse Marion, Chacun Hélène, Farinotti Robert, Barratt Gillian
Laboratoire de Physico-chimie, Pharmacotechnie et Biopharmacie, UMR CNRS 8612, Faculté de Pharmacie, Université Paris-Sud, Châtenay-Malabry, France.
Biochem Pharmacol. 2006 Feb 14;71(4):486-96. doi: 10.1016/j.bcp.2005.11.008. Epub 2005 Dec 6.
Miltefosine (hexadecylphosphocholine, HePC) is the first effective oral agent for the treatment of visceral leishmaniasis. This study aimed to determine whether this oral administration alters the integrity and transport capacities of the intestinal barrier. The objectives of this study were: (i) to evaluate the cytotoxicity of HePC, (ii) to investigate the effects of HePC on paracellular and transcellular transport and (iii) to investigate the influence of HePC on three major transporters of the intestinal barrier, namely, P-glycoprotein, the human intestinal peptide transporter (PepT-1) and the monocarboxylic acid transporter (MCT-1) in Caco-2 cell monolayers, used as an in vitro model of the human intestinal barrier. We show that HePC reduced the transepithelial electrical resistance and increased D-[14C]mannitol permeability in a dose-dependent manner but had no effect on [3H]testosterone permeability, demonstrating that HePC treatment enhances paracellular permeability via an opening of the tight junction complex without affecting the transcellular route. Morphological studies using confocal fluorescence microscopy showed no perturbation of the normal distribution of ZO-1, occludin or E-cadherin but revealed a redistribution of the tight junction-associated protein claudin-1 and the perijunctional actin after incubation with HePC. Finally, HePC was found to inhibit the intestinal P-glycoprotein in the Caco-2 cell model after a single short exposure. These results suggest that HePC could modify the oral bioavailability of other therapeutic compounds absorbed via the paracellular route or which are substrates of the intestinal P-glycoprotein.
米替福新（十六烷基磷酰胆碱，HePC）是首个用于治疗内脏利什曼病的有效口服药物。本研究旨在确定这种口服给药方式是否会改变肠道屏障的完整性和转运能力。本研究的目的是：（i）评估HePC的细胞毒性，（ii）研究HePC对细胞旁转运和跨细胞转运的影响，以及（iii）研究HePC对作为人类肠道屏障体外模型的Caco-2细胞单层中肠道屏障的三种主要转运蛋白，即P-糖蛋白、人类肠道肽转运体（PepT-1）和单羧酸转运体（MCT-1）的影响。我们发现，HePC以剂量依赖的方式降低了跨上皮电阻并增加了D-[14C]甘露醇的通透性，但对[3H]睾酮的通透性没有影响，这表明HePC处理通过打开紧密连接复合物增强了细胞旁通透性，而不影响跨细胞途径。使用共聚焦荧光显微镜进行的形态学研究表明，ZO-1、闭合蛋白或E-钙黏蛋白的正常分布没有受到干扰，但显示与HePC孵育后，紧密连接相关蛋白claudin-1和连接周肌动蛋白发生了重新分布。最后，在单次短暂暴露后，发现HePC在Caco-2细胞模型中抑制肠道P-糖蛋白。这些结果表明，HePC可能会改变其他通过细胞旁途径吸收或作为肠道P-糖蛋白底物的治疗性化合物的口服生物利用度。
