Gender Difference of Hepatic and Intestinal CYP3A4 in CYP3AHumanized Mice Generated by a Human Chromosome-engineering Technique.

Drug metabolism letters Pub Date : 2017-11-17 DOI:10.2174/1872312811666170404153804

Kaoru Kobayashi, Chihiro Abe, Mika Endo, Yasuhiro Kazuki, Mitsuo Oshimura, Kan Chiba

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引用次数: 15

Abstract

Background: Cytochrome P450 3A4 (CYP3A4) is an important drug-metabolizing enzyme that is expressed in the liver and small intestine of humans. Various factors influence the expression of CYP3A4, but gender difference in CYP3A4 expression remains debatable.

Objective: To clarify gender difference of hepatic and intestinal CYP3A4 in CYP3A-humanized mice generated by a human artificial chromosome (HAC) vector system. The CYP3A-humanized (CYP3AHAC) mice have essential regulatory regions, including promoters and enhancers, and unknown elements affecting the expression of CYP3A4.

Methods: We examined the expression and activity of hepatic and intestinal CYP3A4 in male and female CYP3A-HAC mice. CYP3A activity was determined as α- and 4-hydroxylation activity of triazolam in liver and intestinal microsomes. Expression level of CYP3A protein was determined by Western blot analysis. Expression level of CYP3A4 mRNA was measured by quantitative real-time PCR.

Results: The results showed that triazolam hydroxylation activities and protein levels of CYP3A in the liver were significantly higher in female than in male CYP3A-HAC mice, whereas those in the intestine were not significantly different between the genders. In addition, the expression of CYP3A4 mRNA showed a tendency similar to that found for the activity and expression of CYP3A protein in the liver and intestine of CYP3A-HAC mice.

Conclusion: These findings suggest that the expression and activity levels of CYP3A4 in the liver are higher in females than in males, whereas there is no gender difference in the levels in the intestine of CYP3A-HAC mice.

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人染色体工程技术培养的CYP3A4人源化小鼠肝脏和肠道CYP3A4基因的性别差异

背景:细胞色素P450 3A4 (CYP3A4)是人类肝脏和小肠中表达的重要药物代谢酶。多种因素影响CYP3A4的表达，但CYP3A4表达的性别差异仍有争议。目的:探讨人人工染色体(human artificial chromosome, HAC)载体系统培养的cyp3a人源化小鼠肝脏和肠道CYP3A4基因的性别差异。CYP3A4人源化(CYP3AHAC)小鼠具有必要的调控区域，包括启动子和增强子，以及影响CYP3A4表达的未知元素。方法:检测雄性和雌性CYP3A-HAC小鼠肝脏和肠道中CYP3A4的表达和活性。用肝和肠微粒体中三唑仑的α-和4-羟基化活性测定CYP3A活性。Western blot检测CYP3A蛋白表达水平。采用实时荧光定量PCR检测CYP3A4 mRNA的表达水平。结果:雌性CYP3A- hac小鼠肝脏中三唑仑羟基化活性和CYP3A蛋白水平显著高于雄性，而肠道中CYP3A- hac小鼠的三唑仑羟基化活性和蛋白水平在性别间无显著差异。此外，CYP3A4 mRNA的表达与CYP3A- hac小鼠肝脏和肠道中CYP3A蛋白的活性和表达趋势相似。结论:雌性小鼠肝脏中CYP3A4的表达和活性水平高于雄性小鼠，而CYP3A-HAC小鼠肠道中CYP3A4的表达和活性水平无性别差异。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Drug metabolism letters Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science

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期刊介绍： Drug Metabolism Letters publishes letters and research articles on major advances in all areas of drug metabolism and disposition. The emphasis is on publishing quality papers very rapidly by taking full advantage of the Internet technology both for the submission and review of manuscripts. The journal covers the following areas: In vitro systems including CYP-450; enzyme induction and inhibition; drug-drug interactions and enzyme kinetics; pharmacokinetics, toxicokinetics, species scaling and extrapolations; P-glycoprotein and transport carriers; target organ toxicity and interindividual variability; drug metabolism and disposition studies; extrahepatic metabolism; phase I and phase II metabolism; recent developments for the identification of drug metabolites.