INSERM U1059, Dysfonction Vasculaire et Hémostase (DVH), Université Jean Monnet, Saint-Etienne F-42023, France.
INSERM U1059, Laboratoire de Biologie Intégrative du Tissu Osseux (LBTO), Université Jean Monnet, Saint-Etienne F-42023, France.
Mol Pharm. 2022 Jul 4;19(7):2327-2334. doi: 10.1021/acs.molpharmaceut.2c00140. Epub 2022 Jun 8.
Inflammation is characterized by an increased secretion of proinflammatory cytokines known to alter the expression and functionality of drug transporters. Since -glycoprotein (-gp) plays a key role in the pharmacokinetics of several drugs, these modulations could further affect drug exposure. In this context, this study aims to investigate the impact of in vitro cytokine exposure on the expression and activity of -gp using the intestinal model Caco-2 and the human renal cells RPTEC/TERT1. Cells were exposed to various concentrations of tumor necrosis factor (TNF)-α and interleukin (IL)-1β for 24 or 72 h. Gene expression was then assessed by RT-qPCR followed by absolute quantification of -gp using liquid chromatography coupled with mass spectrometry. Then, the activity of -gp was assessed by the intracellular accumulation of rhodamine 123. TNF-α increased both the gene expression and -gp activity by 15-40% in each model. Minor modulations were observed at the protein level with increases of up to 8% for RPTEC/TERT1 cells and 24% for Caco-2 cells. Conversely, IL-1β led to a downregulation of gene, protein, and functionality by 48 and 25% in intestinal and renal cells, respectively. Taken together, these data highlighted that gene expression levels and functional activity of -gp are altered by the pro-inflammatory cytokines in intestinal and renal cells. Such pronounced changes in human -gp could result in altered exposure to drug substrates. Further in vivo studies are needed to confirm the impact of inflammation on drug pharmacokinetics.
炎症的特征是促炎细胞因子的分泌增加，已知这些细胞因子会改变药物转运体的表达和功能。由于β-糖蛋白（β-gp）在几种药物的药代动力学中发挥着关键作用，这些调节可能会进一步影响药物暴露。在这种情况下，本研究旨在使用肠模型 Caco-2 和人肾细胞 RPTEC/TERT1 来研究细胞因子体外暴露对β-gp 表达和活性的影响。细胞分别暴露于不同浓度的肿瘤坏死因子（TNF）-α和白细胞介素（IL）-1β 24 或 72 小时。然后通过 RT-qPCR 评估基因表达，然后使用液相色谱-质谱联用技术对β-gp 进行绝对定量。然后，通过罗丹明 123 的细胞内积累来评估β-gp 的活性。TNF-α在每种模型中使基因表达和β-gp 活性分别增加 15-40%。在蛋白水平上观察到较小的调节，RPTEC/TERT1 细胞增加高达 8%，Caco-2 细胞增加 24%。相反，IL-1β导致基因、蛋白和功能在肠和肾细胞中分别下调 48%和 25%。总之，这些数据表明，促炎细胞因子可改变肠和肾细胞中β-gp 的基因表达水平和功能活性。人β-gp 的这种明显变化可能导致药物底物暴露的改变。需要进一步的体内研究来确认炎症对药物药代动力学的影响。
