酒精对 U937 巨噬细胞中药物外排蛋白和药物代谢酶的影响。
Effect of alcohol on drug efflux protein and drug metabolic enzymes in U937 macrophages.
机构信息
University of Missouri-Kansas City, 64108, USA.
出版信息
Alcohol Clin Exp Res. 2011 Jan;35(1):132-9. doi: 10.1111/j.1530-0277.2010.01330.x. Epub 2010 Oct 6.
BACKGROUND
ATP-binding cassette (ABC) proteins and cytochrome P450 (CYP) enzymes regulate the bioavailability of HIV-1 antiretroviral therapeutic drugs, non-nucleoside reverse transcriptase inhibitors (NNRTIs), and protease inhibitors (PIs). They are also involved in regulating, and responding to, oxidative stress in various tissues and organs including liver. This study is designed to assess the effect of alcohol on the ABCC1 and CYP enzymes involved in the metabolism of NNRTIs and PIs (CYP2B6, CYP2D6, and CYP3A4) and oxidative stress (CYP1A1, CYP2A6, and CYP2E1) in U937 macrophages. The U937 cell line has been utilized as an in vitro model of human macrophages.
METHODS
The expression levels of the ABCC1 and CYP enzymes in U937 macrophages were characterized in terms of mRNA quantification, protein analysis, and assays for functional activity. In addition, oxidative stress was monitored by measuring the activities of oxidative stress marker enzymes and production of reactive oxygen species (ROS).
RESULTS
The order of mRNA expression in U937 macrophages was ABCC1 ∼ CYP2A6 > CYP3A4 ∼ CYP2E1 ∼ CYP1A1 > CYP2D6 > CYP2B6. Alcohol (100 mM) increased the mRNA levels of ABCC1 and CYP2A6 (200%), CYP2B6 and CYP3A4 (150%), and CYP2E1 (400%) compared with the control. Alcohol caused significant upregulation of ABCC1, CYP2A6, CYP2E1, and CYP3A4 proteins (50 to 85%) and showed >50% increase in the specific activity of CYP2A6 and CYP3A4 in U937 macrophages. Furthermore, alcohol increased the production of ROS and significantly enhanced the activity of oxidative stress marker enzymes, superoxide dismutase, and catalase in U937 macrophages.
CONCLUSIONS
Our study showed that alcohol causes increases in the genetic and functional expressions of ABCC1 and CYP enzymes in U937 macrophages. This study has clinical implications in alcoholic HIV-1 individuals, because alcohol consumption is reported to reduce the therapeutic efficacy of NNRTIs and PIs and increases oxidative stress.
背景
三磷酸腺苷结合盒（ABC）蛋白和细胞色素 P450（CYP）酶调节 HIV-1 抗逆转录病毒治疗药物、非核苷类逆转录酶抑制剂（NNRTIs）和蛋白酶抑制剂（PIs）的生物利用度。它们还参与调节和应对包括肝脏在内的各种组织和器官的氧化应激。本研究旨在评估酒精对涉及 NNRTIs 和 PIs（CYP2B6、CYP2D6 和 CYP3A4）代谢和氧化应激（CYP1A1、CYP2A6 和 CYP2E1）的 ABCC1 和 CYP 酶在 U937 巨噬细胞中的作用。U937 细胞系已被用作人类巨噬细胞的体外模型。
方法
通过 mRNA 定量、蛋白分析和功能活性测定，对 U937 巨噬细胞中 ABCC1 和 CYP 酶的表达水平进行了表征。此外，通过测量氧化应激标志物酶的活性和活性氧（ROS）的产生来监测氧化应激。
结果
U937 巨噬细胞中 mRNA 表达的顺序为 ABCC1∼CYP2A6>CYP3A4∼CYP2E1∼CYP1A1>CYP2D6>CYP2B6。与对照组相比，酒精（100mM）使 ABCC1 和 CYP2A6（200%）、CYP2B6 和 CYP3A4（150%）和 CYP2E1（400%）的 mRNA 水平增加。酒精导致 ABCC1、CYP2A6、CYP2E1 和 CYP3A4 蛋白显著上调（50%至 85%），并使 U937 巨噬细胞中 CYP2A6 和 CYP3A4 的特异性活性增加>50%。此外，酒精增加了 ROS 的产生，并显著增强了 U937 巨噬细胞中氧化应激标志物酶超氧化物歧化酶和过氧化氢酶的活性。
结论
我们的研究表明，酒精导致 U937 巨噬细胞中 ABCC1 和 CYP 酶的遗传和功能表达增加。这项研究在酒精性 HIV-1 个体中具有临床意义，因为据报道，酒精消耗会降低 NNRTIs 和 PIs 的治疗效果，并增加氧化应激。
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