RESVERATROL INDUCTION OF GENE EXPRESSION VIA ACTIVATION OF CAR AND NRF2

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 最近，红酒在美国的消费量有所增加，因为流行媒体和医学界都在吹捧红酒的抗氧化特性和对抗动脉粥样硬化和癌症等疾病的潜力。因此，了解白藜芦醇的保护作用并确定合适的啮齿动物模型来研究白藜芦醇对公众健康具有重要意义。在细胞和大鼠肝脏中，白藜芦醇处理增加了受抗氧化反应元件(ARE)调节的几个基因的表达，这些基因对I期和II期新陈代谢至关重要。白藜芦醇发挥保护作用的一个机制是通过激活ARES诱导基因表达。初步研究表明，结构类似于白藜芦醇的二苯乙烯氧化物(TSO)可增加药物代谢酶(DMES)和药物转运蛋白基因的表达。Tso激活两个转录因子，它们调节Dme和转运蛋白的表达，即构造性雄激素受体(CAR)和核因子-E2相关因子(NRF2)。我们假设，白藜芦醇通过激活CAR和NRF2，诱导肝细胞中I期和II期DMES基因以及转运蛋白的表达。具体目标1将确定白藜芦醇治疗是否增加人肝细胞和小鼠肝脏中I期、II期DME和转运蛋白的表达。具体目的2将确定白藜芦醇是否激活CAR和NRF2，使用核受体-1和ARE/EPRE-荧光素酶报告构建的体内荧光素酶检测，以及具有ARE-荧光素酶报告基因的转基因小鼠。具体目标3将确定Res预处理是否影响对乙酰氨基酚的代谢、处置和毒性。总之，这些研究将确定白藜芦醇是否会改变人类以及DMES和转运蛋白基因的表达，以及这些基因的调控是否存在物种相似性。这些研究还将确定白藜芦醇是否通过激活核受体，特别是CAR和NRF2来发挥其保护作用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Recently, red wine consumption in the United States has increased because the popular media and medical community have touted its antioxidant properties and potential to fight disease such as atherosclerosis and cancer. Thus, understanding the protective properties of resveratrol and determining appropriate rodent models to study them is important for public health. In cells and rat liver, resveratrol treatment increases the expression of several genes regulated by the Antioxidant Response Element (ARE), which are important for Phase-I and -II metabolism. A mechanism by which resveratrol may exert its protective properties is through induction of gene expression via activation AREs. Preliminary data demonstrates that trans-stilbene oxide (TSO), a chemical structurally similar to resveratrol, increases the mRNA expression of some genes encoding Phase-I and -II drug metabolizing enzymes (DMEs), and drug transporters. TSO activates two transcription factors that regulate expression of DMEs and transporters, namely the Constitutive Androstane Receptor (CAR) and Nuclear Factor-E2-Related Factor (NRF2). We hypothesize that resveratrol induces the expression of genes for Phase-I, and -II DMEs, as well as transporters, in hepatocytes through activation of CAR and NRF2. Specific aim 1 will determine whether resveratrol treatment increases Phase-I, -II DME expression and transporter expression in human hepatocytes and mouse liver. Specific aim 2 will determine whether resveratrol activates CAR and Nrf2 using transient transfection and in vivo luciferase assays with Nuclear Receptor-1 and ARE/EpRE-luciferase reporter constructs, as well as transgenic mice possessing an ARE-luciferase reporter. Specific aim 3 will determine whether RES pretreatment affects acetaminophen metabolism, disposition, and toxicity. Together, these studies will determine whether resveratrol exposure alters expression of human and genes for DMEs and transporters, and whether a species similarity in regulation of these genes exists. The studies will also determine whether resveratrol exerts its protective properties through activation of nuclear receptors, specifically CAR and Nrf2.