Effect of tumor necrosis factor-α and interferon-γ on intestinal P-glycoprotein expression, activity, and localization in Caco-2 cells
Effect of tumor necrosis factor-α and interferon-γ on intestinal P-glycoprotein expression, activity, and localization in Caco-2 cells
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DOI：
10.1002/jps.20072
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发表时间：
2004-06-01
影响因子：
3.8
通讯作者：
Leroy, C
中科院分区：
文献类型：
--
作者：
Belliard, AM;Lacour, B;Leroy, C
P-糖蛋白（Pgp）是一种药物外排泵，在肠上皮细胞中表达，在那里它构成了对外源性物质的屏障。在炎症性肠病中，已经报道了肿瘤坏死因子（TNF）α和干扰素（IFN）γ的产生失调以及Pgp表达和活性的改变。本研究的目的是研究TNF α和IFN γ对在滤器上生长的Caco-2细胞中肠Pgp表达、活性和定位的影响。TNF α，诱导强烈的时间依赖性减少（-56%）的MDR 1 mRNA（半定量逆转录聚合酶链反应）和显着减少的单向运输的罗丹明123后，48小时的暴露在10 ng/mL。通过共聚焦激光扫描显微镜，Pgp主要定位于对照和TNFa处理的细胞的顶端质膜。相反，IFN γ诱导MDR 1 mRNA和Pgp蛋白表达上调，而不影响Pgp活性。有趣的是，观察到Pgp与横向F-肌动蛋白的共定位。与TNF α相关，IFN γ对Pgp活性既不产生拮抗作用，也不产生协同作用。总之，我们的研究结果表明，TNF α的抑制作用和IFN γ对Pgp转运活性没有影响，使用罗丹明123作为底物。这些细胞因子的作用机制仍有待研究。(C)2004 Wiley-Liss，Inc.和美国药剂师协会
The P-glycoprotein (Pgp), a drug efflux pump, is expressed in intestinal epithelial cells, where it constitutes a barrier against xenobiotics. In inflammatory bowel disease, a dysregulation in the production of tumor necrosis factor (TNF)alpha and interferon (IFN)gamma, and an alteration of Pgp expression and activity have been reported. The aim of this study was to investigate the effects of TNFalpha and IFNgamma on intestinal Pgp expression, activity, and localization in Caco-2 cells grown on filters. TNFalpha, induced both a strong time-dependent diminution (-56%) of MDR1 mRNA (semiquantitative reverse transcription polymerase chain reaction) and a significant decrease of unidirectional transport of rhodamine 123 after 48 h of exposure at 10 ng/mL. By confocal laser scanning microscopy, the Pgp was mainly localized to the apical plasma membrane of both control and TNFalpha-treated cells. By contrast, IFNgamma induced up-regulation of both mRNA MDR1 and Pgp protein expression without incidence on Pgp activity. Interestingly, a colocalization of Pgp with lateral F-actin was observed. Associated with TNFalpha, IFNgamma produced neither an antagonist nor synergistic effect on Pgp activity. In conclusion, our results demonstrate an inhibitory effect of TNFalpha and no effect of IFNgamma on Pgp transport activity using rhodamine 123 as a substrate. Mechanisms of action of these cytokines remain to be studied. (C) 2004 Wiley-Liss, Inc. and the American Pharmacists Association.