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: 8663913
• 负责人: Serrine S Lau
• 金额: $ 8.14万
• 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 2016-01-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8663913
• 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; oxidant stress; podocyte; preconditioning; 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.

描述（由申请人提供）：我们已经表明，在116,16-16-二甲基PGE2（DDM-PGE2）介导的活性氧（ROS）诱导的坏死性/肿瘤性细胞死亡的细胞保护作用。蛋白质组学分析表明，细胞保护与选择数量的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的蛋白78（GRP78）。随后，我们证实了全反击毒酸（ATRA）在体外复制DDM-PGE2介导的细胞保护作用，更重要的是，单剂量ATRA（1 mg/kg，6H预处理）完全保护小鼠免受肾脏缺血/再生（I/R）损伤。此外，在这种治疗剂量下，ATRA诱导NRF2响应性抗氧化剂HO-1和NQO1基因，以及核视黄酸受体RAR？，RAR？2，RAR？2，RAR？2和类黄性素X受体X受体RXR RXR RXR？在肾脏。修订后的应用旨在确定ATRA在体外提供细胞保护的分子机制，以及该细胞保护的该机制在体内概括的程度。我们的中心假设是ATRA诱导的细胞保护是由类似于缺血性预处理的机制介导的。在特定目的1中，我们建议确定ATRA在缺氧/二氧化损伤的体外模型（人肾上皮HK-2细胞）中提供细胞保护的能力，并优化在体内缺血/reperfuson模型中ATRA介导的细胞保护方案（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。类维生素类动物的生物学作用通常是通过与其同源核受体相互作用的，即视黄酸受体（RAR）和类视黄素X受体（RXR）介导的。 RAR和/或RXR参与ATRA介导的细胞保护的程度尚不清楚，并且在I/R的体外和体内模型中，特定的AIM 2是否需要ATRA介导的细胞保护剂是否需要与RAR和/或RXR相互作用。因此，在ROS在疾病中起着重要作用的疾病（例如涉及缺血性再灌注损伤的疾病）中起重要作用的疾病（特定的目标1），并最初表征这种效应的药理学基础（特定目标2），具体目的1和2旨在建立视黄素信号传导作为潜在的治疗干预措施的募集。第三个也是最终的特定目的旨在确定ATRA实现细胞保护的分子机制，每个亚目标都集中在初步研究中已经确定的靶标，因为在细胞保护反应中起着重要作用。具体而言，这些介体是NRF2，GRP78和P38 MAPK，每个MAPK都是缺血预处理的关键介体。因此，特定的目标3将确定（i）抗氧化应激反应的诱导是否取决于NRF2； （ii）ATRA介导的细胞保护需要募集ER（GRP78）介导的应激反应途径； （iii）ATRA募集NRF2取决于p38 MAPK-GRP78相互作用； （iv）通过测试ATRA诱导的NRF2 - / - 小鼠中ATRA诱导的肾脏染色，在体外模型中鉴定出在体外模型中鉴定的细胞保护的机制。当前研究的重要性在于它们在分子和细胞水平上对类维生素性介导的细胞保护的理解的潜力，这些细胞保护在分子和细胞水平上，随后可以提供对在化学诱导组织损伤或缺氧/缺血性缺血性诱导组织损伤期间临床干预有效的新型治疗策略的见解。