Division of Toxicology, Department of Pharmacology, Toxicology and Therapeutics, Showa University School of Pharmacy, Tokyo, Japan (T.A., S.N.); Pharmacological Research Center, Showa University, Tokyo, Japan (T.A., S.N.); and Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Japan (M.Y.)
Division of Toxicology, Department of Pharmacology, Toxicology and Therapeutics, Showa University School of Pharmacy, Tokyo, Japan (T.A., S.N.); Pharmacological Research Center, Showa University, Tokyo, Japan (T.A., S.N.); and Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Japan (M.Y.).
Drug Metab Dispos. 2020 Aug;48(8):673-680. doi: 10.1124/dmd.120.000010. Epub 2020 Jun 5.
NF-E2-related factor 2 (Nrf2) is a transcriptional regulator of biologic defense proteins, such as antioxidant proteins and phase II detoxification enzymes. Cytochrome P450 (P450) enzymes have been shown to regulate phase I metabolism of various drugs and are partially regulated by Nrf2; however, the influence of Nrf2 on drug pharmacokinetics is not known. Here, we showed that Nrf2 depletion prolonged the effect of pentobarbital, a sleep-promoting drug. Pretreatment with phenobarbital, a P450 inducer, shortens the sleeping time associated with pentobarbital-induced sedation in wild-type (WT) mice; however, this effect was not observed in Nrf2 mice. Furthermore, the blood pentobarbital concentration was higher in Nrf2 mice than in WT mice at 30-60 minutes, and the phenobarbital-induced enhancement of its clearance was attenuated in Nrf2 mice compared with WT mice. Total P450 content was decreased in Nrf2 mouse livers, and the phenobarbital-induced increase in P450 content was lower in Nrf2 mice than WT mice. Cyp1a2, Cyp2a5, Cyp2c29, and Cyp2e1 gene expression levels under physiologic conditions and Cyp1a2, Cyp2a5, and Cyp2b10 gene expression levels under phenobarbital-treated conditions were lower in Nrf2 mice compared with WT mice. Additionally, pentobarbital metabolism in liver microsomes was attenuated by Nrf2 depletion. Taken together, these findings suggested that Nrf2 influenced pentobarbital pharmacokinetics through the regulation of drug metabolism and P450 gene expression. Thus, Nrf2-mediated regulation of P450 may contribute to the biologic defense against increased reactive oxygen species production. SIGNIFICANCE STATEMENT: NF-E2-related factor 2 (Nrf2) plays a critical role in the cellular defense against oxidative stress. Nrf2 mice with reduced ability to eliminate reactive oxygen species (ROS) showed a significant delay in emergence from pentobarbital-induced sleep, which was associated with decreased P450 activities and gene expression. Our findings provide that Nrf2 dysfunction or ROS that exceed a threshold level of the eliminating ability of the Nrf2 system may reduce P450 activity.
NF-E2 相关因子 2 (Nrf2) 是生物防御蛋白的转录调节因子，如抗氧化蛋白和 II 相解毒酶。细胞色素 P450（P450）酶已被证明可调节各种药物的 I 相代谢，并且部分受 Nrf2 调节；然而，Nrf2 对药物药代动力学的影响尚不清楚。在这里，我们表明 Nrf2 耗竭延长了戊巴比妥的作用，戊巴比妥是一种促进睡眠的药物。用苯巴比妥预处理，一种 P450 诱导剂，可缩短野生型（WT）小鼠中与戊巴比妥诱导镇静相关的睡眠时间；然而，在 Nrf2 小鼠中未观察到这种效果。此外，在 30-60 分钟时，Nrf2 小鼠血液中的戊巴比妥浓度高于 WT 小鼠，并且与 WT 小鼠相比，Nrf2 小鼠中苯巴比妥诱导的清除增强减弱。Nrf2 小鼠肝脏中的总 P450 含量降低，并且在 Nrf2 小鼠中，苯巴比妥诱导的 P450 含量增加低于 WT 小鼠。在生理条件下，Nrf2 小鼠 Cyp1a2、Cyp2a5、Cyp2c29 和 Cyp2e1 基因表达水平以及在苯巴比妥处理条件下 Cyp1a2、Cyp2a5 和 Cyp2b10 基因表达水平均低于 WT 小鼠。此外，Nrf2 耗竭削弱了肝微粒体中的戊巴比妥代谢。综上所述，这些发现表明 Nrf2 通过调节药物代谢和 P450 基因表达来影响戊巴比妥的药代动力学。因此，Nrf2 介导的 P450 调节可能有助于对增加的活性氧产生的生物防御。意义陈述：NF-E2 相关因子 2 (Nrf2) 在细胞对抗氧化应激中起着至关重要的作用。Nrf2 小鼠的活性氧清除能力降低，表现为从戊巴比妥诱导的睡眠中苏醒明显延迟，这与 P450 活性和基因表达降低有关。我们的研究结果表明，Nrf2 功能障碍或超过 Nrf2 系统消除能力的 ROS 阈值水平可能会降低 P450 活性。
