RESVERATROL INDUCTION OF GENE EXPRESSION VIA ACTIVATION OF CAR AND NRF2

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 美国的红葡萄酒消费量增加，因为大众媒体和医学界都吹捧其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特性，并确定适当的啮齿动物模型来研究它们对公共卫生非常重要。 在细胞和大鼠肝脏中，白藜芦醇治疗增加了由抗氧化反应元件（ARE）调节的几个基因的表达，这对I期和II期代谢很重要。 白藜芦醇可能发挥其保护作用的机制是通过激活战神诱导基因表达。 初步数据表明，反式二苯乙烯氧化物（TSO），一种化学结构类似于白藜芦醇，增加了一些基因编码的第一阶段和第二阶段药物代谢酶（DME）和药物转运蛋白的mRNA表达。 TSO激活调节DME和转运蛋白表达的两种转录因子，即组成型雄烷受体（CAR）和核因子-E2相关因子（NRF 2）。 我们假设白藜芦醇通过激活CAR和NRF 2诱导肝细胞中I期和II期DME以及转运蛋白基因的表达。 具体目标1将确定白藜芦醇处理是否增加人肝细胞和小鼠肝脏中I、II期DME表达和转运蛋白表达。 具体目标2将使用瞬时转染和使用核受体-1和ARE/EpRE-荧光素酶报告基因构建体的体内荧光素酶测定以及具有ARE-荧光素酶报告基因的转基因小鼠来确定白藜芦醇是否激活CAR和Nrf 2。 具体目标3将确定RES预处理是否影响对乙酰氨基酚代谢、处置和毒性。 总之，这些研究将确定白藜芦醇暴露是否会改变人类和DME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Nrf 2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. 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.