Rifampin Regulation of Drug Transporters Gene Expression and the Association of MicroRNAs in Human Hepatocytes.
Rifampin Regulation of Drug Transporters Gene Expression and the Association of MicroRNAs in Human Hepatocytes.
复制标题
DOI：
10.3389/fphar.2016.00111
复制
发表时间：
2016
影响因子：
5.6
通讯作者：
Skaar TC
中科院分区：
文献类型：
--
作者：
Benson EA;Eadon MT;Desta Z;Liu Y;Lin H;Burgess KS;Segar MW;Gaedigk A;Skaar TC
膜药物转运体有助于许多药物的处置。在人类肝脏中，药物转运由两个主要转运蛋白超家族控制，即溶质转运蛋白（SLC）和ATP结合盒转运蛋白（ABC）。由于药物-药物相互作用，这些转运蛋白表达的改变可能导致药物暴露和可能的效果的差异。在这项研究中，我们确定了利福平对数百种膜转运蛋白以及所有临床相关药物转运蛋白基因表达的影响。方法：培养原代人肝细胞（n = 7个供体），用利福平和对照处理24 h。从肝细胞中分离RNA，采用RNA-seq法测定mRNA表达量，采用Taqman OpenArray法分析miRNA表达量。利福平对选定转运蛋白表达的影响也在肾细胞系中进行了测试。比较利福平对19个不同转运基因家族410个转运基因表达的影响。结果：利福平可改变12个临床相关药物转运蛋白基因的表达模式（FDR < 0.05）。例如，ABCC2、ABCB1和ABCC3的表达分别增加了1.9倍、1.7倍和1.2倍。利福平对四种摄取药物转运体（SLCO1B3、SLC47A1、SLC29A1、SLC22A9）的影响与利福平对特异性microRNA表达（SLCO1B3/miR-92a、SLC47A1/miR-95、SLC29A1/miR-30d#和SLC22A9/miR-20）的影响呈负相关，r < - 0.79, p < 0.05)。7种肝脏药物转运基因（SLC22A1、SLC22A5、SLC15A1、SLC29A1、SLCO4C1、ABCC2和ABCC4）在肾近端小管细胞系中也存在，它们在肝细胞中的表达被利福平改变，但在肾细胞中利福平没有改变它们的基因表达。肾细胞中PXR表达极低；这也许可以解释为什么利福平以组织特异性的方式诱导基因表达。结论：利福平改变了许多临床相关的肝脏药物转运体的表达，这可能为理解利福平诱导的体内药物相互作用提供了合理的基础。它对许多其他转运蛋白的影响的相关性仍有待研究。
Membrane drug transporters contribute to the disposition of many drugs. In human liver, drug transport is controlled by two main superfamilies of transporters, the solute carrier transporters (SLC) and the ATP Binding Cassette transporters (ABC). Altered expression of these transporters due to drug-drug interactions can contribute to differences in drug exposure and possibly effect. In this study, we determined the effect of rifampin on gene expression of hundreds of membrane transporters along with all clinically relevant drug transporters. Methods: In this study, primary human hepatocytes (n = 7 donors) were cultured and treated for 24 h with rifampin and vehicle control. RNA was isolated from the hepatocytes, mRNA expression was measured by RNA-seq, and miRNA expression was analyzed by Taqman OpenArray. The effect of rifampin on the expression of selected transporters was also tested in kidney cell lines. The impact of rifampin on the expression of 410 transporter genes from 19 different transporter gene families was compared with vehicle control. Results: Expression patterns of 12 clinically relevant drug transporter genes were changed by rifampin (FDR < 0.05). For example, the expressions of ABCC2, ABCB1, and ABCC3 were increased 1.9-, 1.7-, and 1.2-fold, respectively. The effects of rifampin on four uptake drug transporters (SLCO1B3, SLC47A1, SLC29A1, SLC22A9) were negatively correlated with the rifampin effects on specific microRNA expression (SLCO1B3/miR-92a, SLC47A1/miR-95, SLC29A1/miR-30d#, and SLC22A9/miR-20; r < −0.79; p < 0.05). Seven hepatic drug transporter genes (SLC22A1, SLC22A5, SLC15A1, SLC29A1, SLCO4C1, ABCC2, and ABCC4), whose expression was altered by rifampin in hepatocytes, were also present in a renal proximal tubular cell line, but in renal cells rifampin did not alter their gene expression. PXR expression was very low in the kidney cells; this may explain why rifampin induces gene expression in a tissue-specific manner. Conclusion: Rifampin alters the expression of many of the clinically relevant hepatic drug transporters, which may provide a rational basis for understanding rifampin-induced drug-drug interactions reported in vivo. The relevance of its effect on many other transporters remains to be studied.