Molecular Pharmacology of Insulin Resistance in Burns

烧伤胰岛素抵抗的分子药理学

基本信息

批准号：
7585601
负责人：
Jeevendra Martyn
金额：
$ 39.07万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-01-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7585601
关键词：
1-Phosphatidylinositol 3-Kinase Abbreviations Adenoviruses Anabolism Animals Anti-Inflammatory Agents Anti-inflammatory Area Area Under Curve Attenuated Binding Proteins Biotin Boxing Burn injury Butyric Acid Butyric Acids Carbohydrates Carbon Dioxide Catabolism Cells Conflict (Psychology)Critical Illness Cultured Cells Cyclic GMP Cysteine Data Depressed mood Diabetes Mellitus Diabetic mouse Disruption Dithiothreitol Dose Embryo Endoplasmic Reticulum Endotoxemia Endotoxins Eukaryotic Initiation Factors Exposure to Fatty Liver Fatty acid glycerol esters Fibroblasts Figs - dietary Functional disorder Genes Gluconeogenesis Glucose tolerance test Glycogen Glycogen (Starch) Synthase Glycogen Synthase Kinase 3 Glycogen Synthase Kinases HYOU1 gene Hepatic Hormones Hyperglycemia Immobilization In Vitro Inflammation Inflammation Mediators Inflammatory Insulin Insulin Receptor Insulin Resistance Insulin Signaling Pathway Interferons Interleukins Knock-out Knockout Mice LacZ Genes Leptin Lipopolysaccharides Liver Mass Spectrum Analysis Mediating Mediation Mediator of activation protein Metabolic Methods Molecular Molecular Chaperones Morbidity - disease rate Mus Muscle Muscle Cells Muscle Fibers Muscle Proteins N-acetylpenicillamine Neurons Nitric Oxide Nitric Oxide Synthase Nitric Oxide Synthase Type I Non-Insulin-Dependent Diabetes Mellitus Nuclear Obesity Oxidation-Reduction Oxidoreductase Pancreas Pathogenesis Pathologic Pharmacology Pharmacotherapy Phosphate Buffer Phosphoenolpyruvate Carboxylase Phosphotransferases Play Post-Translational Protein Processing Production Protein Biosynthesis Protein Overexpression Protein Synthesis Inhibition Proteins Proteomics Proto-Oncogene Proteins c-akt Public Health RNA Reaction Reactive Nitrogen Species Reduced Glutathione Reporting Research Resistance Respiratory Failure Reticulum Right-On Rodent Role S-nitro-N-acetylpenicillamine SKIL gene Saline Signal Pathway Signal Transduction Signaling Protein Skeletal Muscle Skeletal system Small Interfering RNA Solid Stress Substrate Cycling Sulfhydryl Compounds Taurine Testing Text Therapeutic Intervention Thinking Tissues Transgenic Mice Transgenic Organisms Trauma Tumor Necrosis Factor-alpha Tumor Necrosis Factors Ubiquitin Upstream Enhancer Ursodeoxycholic Acid Wild Type Mouse Work aminoguanidine attenuation base biological adaptation to stress cell type cytokine diabetic eIF-2 Kinase glucose uptake glucose-regulated proteins glycemic control human NOS2A protein human NOS3 protein human TNF protein human TYRP1 protein human disease improved in vivo inhibitor/antagonist insulin receptor substrate 1 protein insulin sensitivity insulin signaling insulin tolerance macrophage mortality mouse S-nitrosoglutathione reductase novel therapeutics oxygen-regulated proteins pre-clinical prevent protective effect prototype response stress-activated protein kinase 1 tauroursodeoxycholic acid transcription factor transcription factor CHOP wasting

项目摘要

DESCRIPTION (provided by applicant): Insulin resistance is the major metabolic abnormality associated with burn injury. All insulin-mediated effects, including glucose uptake in tissues, protein synthesis, inhibition of gluconeogenesis and anti-inflammatory functions, are markedly attenuated. Supraphysiologic doses of exogenous insulin to counter burn-induced insulin resistance produce deleterious effects, including increased CO2 production and hepatic steatosis. Burn injury alters the insulin signaling pathway at multiple points, via the insulin receptor, insulin receptor substrate-1 (IRS-1), phosphatidylinositol 3-kinase (PI3-K), Akt/PKB (protein kinase B), and glycogen synthase kinase-32 (GSK-32). Although inducible nitric oxide synthase (iNOS) is thought to play an important role in the deranged insulin-signaling, the molecular mechanisms by which iNOS mediates these changes are unknown. Specific Aim 1 will test the hypothesis that the de-nitrosylation reaction, regulated by S-nitrosoglutathione reductase (GSNOR), plays an important protective role in burn-induced insulin resistance in mice. It is hypothesized that nitrosative stress by iNOS leads to increased protein S-nitrosylation (post-translational modification) of the insulin-signaling proteins resulting in depressed insulin signaling. To test this hypothesis, insulin sensitivity, glucose uptake in muscle, insulin signaling, and S-nitrosylation (by proteomics) of the insulin-signaling proteins will be evaluated in muscle from sham-burned or burned wild-type, GSNOR knockout (-/-) and GSNOR/ iNOS- /- double knockout mice. The protective role of GSK-32 inhibitors in obesity-induced insulin resistance is established, but the molecular mechanism of GSK-32 activation and the salutary effects of GSK-32 inhibitors remain to be investigated particularly in skeletal muscle. Specific Aim 2 will test the hypothesis that (1) iNOS- mediated increased activity of GSK-32 plays an important role in burn-induced insulin resistance; (2) S- nitrosylation is involved in iNOS-mediated GSK-32 activation after burn injury; and (3) GSK-32 activation reduces IRS-1 expression as a downstream effector of iNOS in skeletal muscle. Specific Aim 3 will test the hypothesis that endoplasmic reticulum (ER) stress response plays an important role in muscle insulin resistance of burns, and that iNOS functions both as a downstream effector and an upstream enhancer of ER stress in skeletal muscle. The planned studies will use: XBP-1 , skeletal muscle-specific ORP150 over- expressing transgenic, GSNOR-/-, iNOS-/- and GSNOR/iNOS-/- double knockout mice to test these hypotheses. (XBP-1 is a transcription factor regulating ER chaperones and ORP150 protects cells from ER stress.) Thus, these studies will apply an integrated molecular, pharmacologic, and proteomic approach to elucidate the molecular mechanism by which iNOS, GSNOR, GSK-32, and ER stress interrelate to produce insulin resistance, and will provide a rationale and preclinical data for novel therapeutic interventions to treat insulin resistance in muscle after burns. PUBLIC HEALTH RELEVANCE: The proposed studies will apply integrated molecular, pharmacologic, and proteomic approaches to elucidate the molecular mechanism by which inducible nitric oxide and endoplasmic stress reticulum interact with each other and insulin signaling proteins to cause insulin resistance in burns

描述（由申请人提供）：胰岛素抵抗是与烧伤相关的主要代谢异常。所有胰岛素介导的作用，包括组织中的葡萄糖摄取、蛋白质合成、糖异生抑制和抗炎功能，均显着减弱。使用超生理剂量的外源性胰岛素来对抗烧伤引起的胰岛素抵抗会产生有害影响，包括增加二氧化碳产生和肝脏脂肪变性。烧伤通过胰岛素受体、胰岛素受体底物 1 (IRS-1)、磷脂酰肌醇 3-激酶 (PI3-K)、Akt/PKB（蛋白激酶 B）和糖原合酶激酶在多个点改变胰岛素信号传导通路-32（GSK-32）。尽管诱导型一氧化氮合酶 (iNOS) 被认为在胰岛素信号紊乱中发挥重要作用，但 iNOS 介导这些变化的分子机制尚不清楚。具体目标 1 将检验以下假设：由 S-亚硝基谷胱甘肽还原酶 (GSNOR) 调节的去亚硝基化反应在小鼠烧伤诱导的胰岛素抵抗中发挥重要的保护作用。据推测，iNOS 的亚硝化应激会导致胰岛素信号蛋白的 S-亚硝基化（翻译后修饰）增加，从而导致胰岛素信号传导抑制。为了检验这一假设，将在假烧伤或烧伤野生型、GSNOR 敲除（-/ -) 和 GSNOR/ iNOS- /- 双敲除小鼠。 GSK-32 抑制剂在肥胖引起的胰岛素抵抗中的保护作用已确立，但 GSK-32 激活的分子机制和 GSK-32 抑制剂的有益作用仍有待研究，特别是在骨骼肌中。具体目标 2 将检验以下假设：(1) iNOS 介导的 GSK-32 活性增加在烧伤诱导的胰岛素抵抗中发挥重要作用； (2) S-亚硝基化参与烧伤后iNOS介导的GSK-32激活； (3) GSK-32 激活降低骨骼肌中 iNOS 下游效应子 IRS-1 的表达。具体目标 3 将检验以下假设：内质网 (ER) 应激反应在烧伤的肌肉胰岛素抵抗中发挥重要作用，并且 iNOS 既充当骨骼肌中 ER 应激的下游效应子又充当上游增强子。计划的研究将使用：XBP-1、骨骼肌特异性ORP150过表达转基因、GSNOR-/-、iNOS-/-和GSNOR/iNOS-/-双敲除小鼠来测试这些假设。（XBP-1 是调节 ER 伴侣的转录因子，ORP150 保护细胞免受 ER 应激。）因此，这些研究将应用综合分子、药理学和蛋白质组学方法来阐明 iNOS、GSNOR、GSK-32、和 ER 应激相互关联，产生胰岛素抵抗，并将为治疗烧伤后肌肉胰岛素抵抗的新型治疗干预措施提供基本原理和临床前数据。公共健康相关性：拟议的研究将应用综合分子、药理学和蛋白质组学方法来阐明诱导型一氧化氮和内质应激网彼此相互作用以及胰岛素信号蛋白相互作用导致烧伤中胰岛素抵抗的分子机制