Retinoid Mediated Protection Against Reactive Oxygen Species Induced Cytotoxicity

类维生素A介导的针对活性氧诱导的细胞毒性的保护

• 批准号: 7985510
• 负责人: Serrine S Lau
• 金额: $ 33.8万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7985510
• 关键词: Actins; Acute; Acute Kidney Failure; Adverse effects; Agonist; Animal Model; Animals; Antioxidants; Apoptosis; Attenuated; Biological; Biomedical Research; Brain Hypoxia-Ischemia; Cell Death; Cell Hypoxia; Cell Line; Cell Proliferation; Cell model; Cells; Chemicals; Chronic Kidney Failure; Clinical; Cytoprotection; Data; Development; Dinoprostone; Disease; Dose; Enzymes; Epithelial; Epithelial Cells; Event; Failure; Family; Fibroblasts; Genes; Goals; Human; Hyperlipidemia; Hypoxia; In Vitro; Injection of therapeutic agent; Injury; Intervention; Investigation; Ischemia; Ischemic Preconditioning; Kidney; Kidney Diseases; Knowledge; Lymphocyte; MAP Kinase Gene; MAPK14 gene; Mediating; Mediator of activation protein; Miniature Swine; Modeling; Molecular; Mus; NQO1 gene; Necrosis; Nuclear; Nuclear Hormone Receptors; Nuclear Receptors; Nuclear Translocation; Organ; Oxidation-Reduction; Oxidative Stress; Pathology; Pathway interactions; Play; Process; Proteins; Proteomics; Protocols documentation; Proximal Kidney Tubules; RXR; Reaction; Reactive Oxygen Species; Recruitment Activity; Renal function; Reperfusion Injury; Reperfusion Therapy; Retinoic Acid Receptor; Retinoids; Retinol Binding Proteins; Rhabdomyolysis; Role; Scheme; Signal Pathway; Signal Transduction; Stress; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Thromboxane A2; Thromboxane Receptor; Tissues; Translating; Tretinoin; Tubular formation; Up-Regulation; Vasoconstrictor Agents; Vitamin A; analog; base; biological adaptation to stress; cell injury; cell type; clinically significant; cytotoxicity; design; disulfide bond; glucose-regulated proteins; in vitro Model; in vivo; in vivo Model; insight; interstitial; kidney cell; macrophage; mesangial cell; mortality; novel therapeutics; numb protein; oxidant stress; podocyte; preconditioning; progesterone 11-hemisuccinate-(2-iodohistamine); protective effect; protein misfolding; public health relevance; receptor; receptor function; renal ischemia; repaired; response

DESCRIPTION (provided by applicant): We have shown that retinoid signaling is engaged during 11-deoxy-16,16-dimethyl PGE2 (DDM-PGE2) mediated cytoprotection against reactive oxygen species (ROS) induced necrotic/oncotic cell death. Proteomics analyses revealed that cytoprotection is associated with the increased synthesis of a select number of proteins, including retinol binding protein (RBP), actin, and glucose-regulated protein 78 (Grp78). We subsequently confirmed that all-trans-retinoic acid (aTRA) replicates DDM-PGE2-mediated cytoprotection in vitro, and more importantly, a single dose of aTRA (1 mg/kg, 6h pretreatment) completely protects mice from renal ischemia/reperfusion (I/R) injury. Furthermore, at this therapeutic dose, aTRA induces Nrf2-responsive antioxidant HO-1 and NQO1 genes, as well as nuclear retinoic acid receptors RAR?, RAR?2, RAR?2, and retinoid X receptors RXR? in the kidney. The revised application is designed to determine the molecular mechanisms by which aTRA affords cytoprotection in vitro, and the extent to which this mechanism(s) of cytoprotection is recapitulated in vivo. Our central hypothesis is that aTRA-induced cytoprotection is mediated by mechanisms similar to ischemic preconditioning. In Specific Aim 1 we propose to determine the ability of aTRA to offer cytoprotection in an in vitro model (human renal epithelial HK-2 cells) of hypoxia/reoxygenation injury, and to optimize protocols for aTRA-mediated cytoprotection in an in vivo ischemia/reperfuson model (IR). We will also ascertain whether the protective effects are mediated, at least in part, via the upregulation of anti-oxidant enzymes. The biological effects of retinoids are typically mediated via interaction with their cognate nuclear receptors, namely, retinoic acid receptors (RAR) and retinoid X receptors (RXR). The extent to which RAR and/or RXR participate in aTRA-mediated cytoprotection is not known, and Specific Aim 2 will determine, in both the in vitro and in vivo models of I/R, whether aTRA-mediated cytoprotection requires interaction with RAR and/or RXR. Specific Aims 1 and 2 are therefore designed to establish the recruitment of retinoid signaling as a potential therapeutic intervention in conditions where ROS play an important role in the pathology of the disease, such as those involving ischemia reperfusion injury (Specific Aim 1), and to initially characterize the pharmacological basis of this effect (Specific Aim 2). The third and final Specific Aim is designed to identify the molecular mechanisms by which aTRA accomplishes cytoprotection, with each sub- aim focusing on a target that has already been identified in preliminary studies as playing an important role in the cytoprotective response. Specifically, those mediators are Nrf2, Grp78, and p38 MAPK, each of which is also a key mediator of ischemia preconditioning. Specific Aim 3 will therefore determine whether (i) aTRA- mediated induction of the anti-oxidant stress response is dependent upon Nrf2; (ii) aTRA-mediated cytoprotection requires the recruitment of the ER (Grp78) mediated stress response pathway; (iii) the recruitment of Nrf2 by aTRA is dependent on p38 MAPK-Grp78 interactions; and (iv) the mechanism(s) of cytoprotection identified in the in vitro model are recapitulated in the in vivo model by testing aTRA induced renoprotection in Nrf2-/- mice. The significance of the current studies resides in their potential to enhance our understanding of retinoid mediated cytoprotection at the molecular and cellular level, which can subsequently provide insights into novel therapeutic strategies effective for clinical interventions during chemical induced tissue injury or hypoxia/ischemia-reperfusion injury. PUBLIC HEALTH RELEVANCE: Oxidative stress has been shown to cause or contribute to many diseases including different forms of renal disease such as acute renal failure, rhabdomyolysis, obstructive nephropathy, hyperlipidemia, glomerular damage, chronic renal failure, and ischemia/reperfusion. An understanding of the factors that regulate the cellular response to oxidative stress and of the molecular mechanisms, by which they interact with cellular constituents, and the consequences of such interactions, remain important fundamental goals of biomedical research. All-trans-retinoic acid (aTRA) may offer an alternative therapeutic strategy offering renoprotection through mechanisms similar to ischemia preconditioning, and therefore studies on the mechanism by which aTRA confers cytoprotection could have important therapeutic implications. Assessment of the beneficial effects of retinoids in animal model and human renal disease warrants further investigation.

描述（由申请人提供）：我们已经表明，类维生素A信号传导在11-脱氧-16,16-二甲基PGE2（DDM-PGE2）介导的细胞保护作用中参与，以抵抗活性氧（ROS）诱导的坏死/胶体细胞死亡。蛋白质组学分析表明，细胞保护与某些蛋白质的合成增加有关，包括视黄醇结合蛋白 (RBP)、肌动蛋白和葡萄糖调节蛋白 78 (Grp78)。我们随后证实，全反式视黄酸（aTRA）在体外复制了 DDM-PGE2 介导的细胞保护作用，更重要的是，单剂量的 aTRA（1 mg/kg，6 小时预处理）完全保护小鼠免受肾缺血/再灌注的影响。 I/R) 损伤。此外，在该治疗剂量下，aTRA 诱导 Nrf2 响应的抗氧化剂 HO-1 和 NQO1 基因，以及核视黄酸受体 RAR?、RAR?2、RAR?2 和类视黄醇 X 受体 RXR?在肾脏。修订后的申请旨在确定 aTRA 在体外提供细胞保护的分子机制，以及这种细胞保护机制在体内重现的程度。我们的中心假设是 aTRA 诱导的细胞保护是通过类似于缺血预处理的机制介导的。在具体目标 1 中，我们建议确定 aTRA 在缺氧/复氧损伤的体外模型（人肾上皮 HK-2 细胞）中提供细胞保护的能力，并优化 aTRA 介导的体内缺血/再灌注模型（IR）。我们还将确定保护作用是否至少部分是通过抗氧化酶的上调来介导的。类视黄醇的生物效应通常通过与其同源核受体（即视黄酸受体（RAR）和类视黄醇X受体（RXR））的相互作用来介导。 RAR 和/或 RXR 参与 aTRA 介导的细胞保护的程度尚不清楚，具体目标 2 将确定在 I/R 的体外和体内模型中，aTRA 介导的细胞保护是否需要与 RAR 和 RXR 相互作用。 /或RXR。因此，具体目标 1 和 2 旨在建立类维生素A信号传导的募集，作为 ROS 在疾病病理学中发挥重要作用的情况下的潜在治疗干预措施，例如涉及缺血再灌注损伤的情况（具体目标 1），并初步描述该效应的药理学基础（具体目标 2）。第三个也是最后一个具体目标旨在确定 aTRA 实现细胞保护的分子机制，每个子目标都集中在初步研究中已确定在细胞保护反应中发挥重要作用的靶标。具体来说，这些介质是 Nrf2、Grp78 和 p38 MAPK，它们都是缺血预处理的关键介质。因此，具体目标 3 将确定 (i) aTRA 介导的抗氧化应激反应的诱导是否依赖于 Nrf2； (ii) aTRA 介导的细胞保护需要招募 ER (Grp78) 介导的应激反应途径； (iii) aTRA 招募 Nrf2 取决于 p38 MAPK-Grp78 相互作用； (iv) 通过在 Nrf2-/- 小鼠中测试 aTRA 诱导的肾脏保护作用，在体内模型中重现体外模型中确定的细胞保护机制。当前研究的意义在于它们有可能增强我们对类维生素A介导的细胞保护在分子和细胞水平上的理解，从而可以为化学诱导的组织损伤或缺氧/缺血再灌注损伤期间有效的临床干预提供新的治疗策略。 。 公共健康相关性：氧化应激已被证明可引起或促成许多疾病，包括不同形式的肾脏疾病，如急性肾衰竭、横纹肌溶解症、阻塞性肾病、高脂血症、肾小球损伤、慢性肾衰竭和缺血/再灌注。了解调节细胞对氧化应激反应的因素以及它们与细胞成分相互作用的分子机制，以及这种相互作用的后果，仍然是生物医学研究的重要基本目标。全反式视黄酸（aTRA）可能提供一种替代治疗策略，通过类似于缺血预处理的机制提供肾脏保护，因此对 aTRA 赋予细胞保护作用的机制的研究可能具有重要的治疗意义。评估类维生素A在动物模型和人类肾脏疾病中的有益作用值得进一步研究。