Project 3: Anti-Inflammatory Mechanisms of Inhaled Hypertonic Saline

项目3：吸入高渗盐水的抗炎机制

• 批准号: 8382283
• 负责人: ANIRBAN BANERJEE
• 金额: $ 36.63万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8382283
• 关键词: Acute; Adhesions; Adhesives; Affect; Aftercare; Alveolar; Alveolar Cell; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Asses; Attenuated; Automobile Driving; Bioinformatics; Blood; Breathing; CCAAT-Enhancer-Binding Proteins; Cell Nucleus; Cell model; Cells; Clathrin; Complex; Cystic Fibrosis; Data; Databases; Development; Exons; Family; Family member; Fluorescence Resonance Energy Transfer; Gene Expression; Genes; Genetic Transcription; Genome; Goals; Hemorrhage; Human; IRF1 gene; Immunoprecipitation; Importins; Inflammatory; Infusion procedures; Interleukin-1; Kinetics; Leukocytes; Libraries; Lung; Lung Inflammation; NF-kappa B; Nuclear Import; Nuclear Translocation; Patients; Peptides; Phenotype; Principal Investigator; Production; Proteins; Proteomics; RNA Interference; Receptor Signaling; Regulation; Reporter; Resuscitation; Reverse Transcriptase Polymerase Chain Reaction; Safety; Saline; Scheme; Shock; Signal Transduction; Site; Stimulus; Testing; Therapeutic; Time; Translating; Trauma; cell type; chemokine; cytokine; density; dimer; dosage; lung injury; macrophage; member; molecular phenotype; neutrophil; novel; p65; pneumocyte; prevent; programs; promoter; receptor; research study; trafficking; transcription factor

Trauma, hemorrhage and blood resuscitation produce acute lung inflammation by inducing chemokine, cytokine and, and adhesive proteins. Up-regulating mechanisms depend on systemic signals that translocate gene-regulating, transcription factors (TFs), such as NF-kB into the nucleus. The genes expressed direct the adhesion, transmigration of leukocytes and phenotypic developments in the alveolar milieu. Hyperosmolarity (HOsm), appears to prevent inflammatory gene expression but the mechanisms and scope are unclear. Therapeutic hypertonic saline infusions during trauma-resuscitation, or inhaled as for cystic fibrosis, are well tolerated. We hypothesize that hyperosmolarity alters the nuclear translocation of selected transcription factors induced by inflammatory stimuli by promoting novel complexes. In this proposal period (2005-2010) we will first examine the traffic of TFs controlling prototypical alveolar cytokines and adhesive proteins that are modified by HOsm, and investigate complexes of these TFs, including those formed with importins. 1. Investigate HOsm altered translocation of the NF-KB and IKK family members, their HOsm altered cytoplasmic complexes and examine alternate complexes promoted by HOsm with IP-MS proteomics. 2. Since HOsm does not prohibit p65 Rel A translocation induced by IL-1 yet suppresses certain cytokines, we will examine translocation of other essential promoters of the IRF, C/EBP. and R/FLAT families and identify their HOsm altered partners. Next, to expand the inquiry beyond the known cytokines we will execute: 3. Gene array analyses, to guide bioinformatics queries for potential TFs. The goal is to gradually build up a library of TFs that do not translocate successfully during HOSm and identify their sequestering partners, (starting with NF-kB and IRF family members). Lastly, we will translate these bench experiments to asses the tolerability or benefit of nebulized HOsm in shocked animals and trauma patients. 4. Evaluate lung inflammation and TF translocation in traumatized Animal or Patients treated with inhaled HOsm. The information from Aims 1-3 will be used to interpret the molecular phenotypes and TF redistributions after treatments.

创伤、出血和血液复苏通过诱导趋化因子、细胞因子和粘附蛋白产生急性肺炎症。上调机制取决于将基因调节转录因子（TF）（例如NF-kB）转移到细胞核中的系统信号。表达的基因指导白细胞在肺泡环境中的粘附、移行和表型发育。高渗透压（HOsm）似乎可以阻止炎症基因的表达，但其机制和范围尚不清楚。创伤复苏期间治疗性高渗盐水输注或吸入治疗囊性纤维化耐受性良好。我们推测，高渗改变核转位选定的转录因子诱导的炎症刺激，促进新的复合物。 在这个提案期间（2005-2010年），我们将首先检查交通的TF控制原型肺泡细胞因子和粘附蛋白的HOsm修改，并调查这些TF的复合物，包括那些形成的importins。 1.研究HOsm改变的NF-κ B和IKK家族成员的易位，其HOsm改变的细胞质复合物，并使用IP-MS蛋白质组学检查HOsm促进的替代复合物。 2.由于HOsm不抑制IL-1诱导的p65 Rel A易位，但抑制某些细胞因子，我们将研究IRF的其他必需启动子C/EBP的易位。和R/FLAT家族，并鉴定其HOsm改变的伴侣。 接下来，为了将调查扩展到已知的细胞因子之外，我们将执行： 3.基因阵列分析，以指导潜在TF的生物信息学查询。我们的目标是逐步建立一个在HOSm过程中不能成功易位的TF库，并鉴定它们的螯合伴侣（从NF-kB和IRF家族成员开始）。 最后，我们将把这些实验室实验转化为评估雾化HOsm在休克动物和创伤患者中的耐受性或益处。 4.评估吸入HOsm治疗创伤动物或患者的肺部炎症和TF易位。目标1-3的信息将用于解释治疗后的分子表型和TF再分布。