Vitamin E deficiency: oxidant induced gene expression

描述(由申请人提供) 这项研究的长期目标是确定分子机制 膳食抗氧化剂与细胞和蛋白质的潜在相互作用 环境氧化剂在体内应用的全局基因表达 分析。选择性缺失的转基因小鼠的可用性 α-生育酚(a-TC)转移蛋白基因及其引起的耗竭 组织中的a-TC(维生素E)，提供了一个独特的机会来严格检查 一种主要的饮食抗氧化剂生物膜和 正常生理中的脂蛋白。此外，它作为一个潜在的 膳食补充剂对慢性病发病机制的干预 由氧化剂引发的污染物在环境中也可以检测到。这个 这项建议的具体目的是：(1)定义和比较 α-生育酚转移蛋白在肺和肝脏中的表达谱 基因敲除(TTP-KO)和野生型小鼠；(2)检测基因表达的变化 肺和肝脏对光化学产生的辐射的响应 TTP-KO和野生型小鼠的环境氧化剂臭氧；以及(3)评估 由选定的mRNA编码的蛋白质表达的变化， 通过特定的目标1和2，通过免疫印迹和免疫- 组织化学分析。 这项拟议的研究将首次确定基因组反应。 体内沉默的基因，其产物在 膜脂蛋白中主要抗氧化剂的动态平衡。此外, 全球基因表达分析也将确定a-a-的分子靶点 TC可能与清除活性氧没有直接关系 TTP的物种和对耗竭的分子反应没有直接关系 A-TC的转移。接触苯系物后的基因表达分析 环境氧化剂臭氧，其主要目标是膜环境， 将确定体内对氧化应激的“转录”反应 由于环境污染物和饮食抗氧化剂的作用， 维生素E对氧化剂诱导的靶基因表达变化的影响 纸巾。对选定的mRNAs编码的蛋白质的分析将建立 检测到的mRNAs变化的潜在功能含义 用基因芯片进行分析。这些分子数据可能为 设计更有效的抗氧化剂饮食干预方案 这可能会缓解环境氧化剂引发的肺部疾病。

DESCRIPTION (provided by applicant) The long-term objectives of this research are to define molecular mechanisms underlying interactions of dietary antioxidants with cellular and environmental oxidants in vivo by the application of global gene expression analysis. The availability of transgenic mice with selective deletion of the gene for alpha-tocopherol (a-TC) transfer protein, and the resulting depletion of tissue a-TC (vitamin E), offers a unique opportunity to rigorously examine the role of a major dietary antioxidant of biological membranes and lipoproteins in normal physiology. In addition, its potential role as a dietary supplement to intervene in the pathogenesis of chronic diseases triggered by oxidant pollutants in the environment can also be tested. The specific aims of this proposal are to: (1) define and compare the mRNA expression profiles of lungs and livers from a-tocopherol transfer protein knockout (TTP-KO) and wild type mice; (2) assay the changes in mRNA expression profiles of lungs and livers in response to a photochemically-generated environmental oxidant, ozone, in TTP-KO and wild type mice; and (3) evaluate the changes in the expression of proteins encoded by selected mRNAs, identified through specific aims 1 and 2, by immunoblot and immuno- histochemical analysis. The proposed research will identify, for the first time, the genomic response in vivo to silencing of a gene whose product plays a significant role in the homeostasis of a primary antioxidant in membranes lipoproteins. In addition, global gene expression analysis will also identify the molecular targets of a- TC that may not be directly related to the scavenging of reactive oxygen species and the molecular response to depletion of TTP not directly related to the transfer of a-TC. Gene expression analysis following exposure to an environmental oxidant, ozone, whose primary target is the membrane milieu, will identify the "transcriptional" response in vivo to oxidative stress inflicted by an environmental pollutant and the role of a dietary antioxidant, vitamin E, on oxidant-induced changes in gene expression in the target tissues. The analysis of proteins encoded by selected mRNAs will establish the potential functional implications of changes in the mRNAs detected by analysis with gene arrays. These molecular data may provide the basis for designing more effective protocols for dietary interventions with antioxidants that might ameliorate lung diseases induced by environmental oxidants.