Department of Pharmacology and Therapeutics, The University of Liverpool, Pembroke Place, Liverpool, UK.
Acta Pharmacol Sin. 2010 Jan;31(1):51-65. doi: 10.1038/aps.2009.187.
The aim of this study was to investigate the impact on expression of mRNA and protein by paradigm inducers/activators of nuclear receptors and their target genes in rat hepatic and intestinal cells. Furthermore, assess marked inter laboratory conflicting reports regarding species and tissue differences in expression to gain further insight and rationalise previously observed species differences between rodent and human based systems.
Quantitative real time-polymerase chain reaction (QRT-PCR) and immunoblots were used to assess messenger RNA (mRNA) and protein expression for CYP2B2, CYP3A1, CYP3A2, CYP3A9, ABCB1a, ABCB1b, ABCC1, ABCC2, pregnane X receptor (PXR), farnesoid X receptor (FXR) and constituitive androstane receptor (CAR) in rat hepatoma cell line H411E, intestinal cells, Iec-6, and rat primary hepatocytes, in response to exposure for 18 h with prototypical inducers.
Dexamethasone (DEX) and pregnenolone 16alpha carbonitrile (PCN) significantly induced PXR, CYP3A9, ABCB1a and ABCB1b. However, when co-incubated, DEX appeared to restrict PCN-dependent induction. Chenodeoxycholic acid (CDCA) was the only ligand to induce FXR in all three cell types. Despite previously reported species differences between PCN and rifampicin (RIF), both compounds exhibited a similar profile of induction.
Data presented herein may explain some of the discrepancies previously reported with respect to species differences from different laboratories and have important implications for study design.
本研究旨在探讨核受体的模式诱导剂/激活剂及其靶基因对大鼠肝肠细胞中 mRNA 和蛋白质表达的影响。此外，评估关于物种和组织表达差异的明显的实验室间相互矛盾的报告，以深入了解和合理化以前观察到的啮齿动物和人类基于系统之间的物种差异。
使用定量实时聚合酶链反应（QRT-PCR）和免疫印迹法评估 CYP2B2、CYP3A1、CYP3A2、CYP3A9、ABCB1a、ABCB1b、ABCC1、ABCC2、孕烷 X 受体（PXR）、法尼醇 X 受体（FXR）和组成型雄烷受体（CAR）在大鼠肝癌细胞系 H411E、肠细胞 Iec-6 和大鼠原代肝细胞中的信使 RNA（mRNA）和蛋白表达，以响应暴露于原型诱导剂 18 小时。
地塞米松（DEX）和孕烯醇酮 16α-腈（PCN）显著诱导了 PXR、CYP3A9、ABCB1a 和 ABCB1b。然而，当共同孵育时，DEX 似乎限制了 PCN 依赖性诱导。鹅去氧胆酸（CDCA）是唯一能诱导三种细胞类型中 FXR 的配体。尽管以前报道过 PCN 和利福平（RIF）之间的物种差异，但这两种化合物表现出相似的诱导谱。
本文提供的数据可以解释一些以前不同实验室报告的物种差异的差异，并对研究设计具有重要意义。
