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Role of heat shock protein 90 in alcoholic liver disease

热休克蛋白 90 在酒精性肝病中的作用

基本信息

批准号：
8497550
负责人：
Pranoti Mandrekar
金额：
$ 34.57万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8497550
关键词：
17-(Allylamino)-17-demethoxygeldanamycin 17-(Dimethylaminoethylamino)-17-Demethoxygeldanamycin Ablation Acetylation Affect Alcohol consumption Alcoholic Liver Diseases Alcohols Binding Chronic Cirrhosis Development Drug Design Drug Targeting Exposure to Fatty Liver Functional disorder HSF1 Health Heat Stress Disorders Heat shock proteins Heat-Shock Proteins 90 Hepatic Inflammation Inflammatory Inflammatory Response Injury to Liver Knowledge Kupffer Cells Link Liver Macrophage Activation Malignant Neoplasms Measures Mediating Molecular Chaperones Morbidity - disease rate Mus NADPH Oxidase Oxidation-Reduction Oxidative Stress Pathogenesis Pathway interactions Phosphotransferases Play Predisposition Primary carcinoma of the liver cells Production Role STAT1 gene Signal Pathway Signal Transduction Signaling Molecule Small Interfering RNA Sp1 Transcription Factor Steatohepatitis Stream Stress TLR4 gene TNF gene Testing alcohol exposure base cell injury chromatin immunoprecipitation chromatin remodeling cytokine dimer feeding heat shock transcription factor human IRAK1 protein inhibitor/antagonist liver injury macrophage mortality novel problem drinker promoter transcription factor

项目摘要

. ABSTRACT Activation of liver resident macrophages and increased pro-inflammatory cytokine production is a hallmark in the pathogenesis of alcoholic liver disease (ALD). Alcohol-induced oxidative stress plays an important part in macrophage activation. Heat shock proteins are induced by oxidative stress and function as molecular chaperones. Heat shock protein 90 (hsp90) and Grp94/gp96 chaperone signaling molecules of the TLR4 pathway to regulate inflammatory cytokines. Thus, hsp90 and gp96 could link stress and inflammatory pathways and play an important role in development of ALD. Our preliminary studies show that chronic alcohol feeding in mice increases hsp90 and gp96 in isolated Kupffer cells (KCs), compared to pair-fed controls. Hsp90 from chronic alcohol-exposed macrophages associates with IKKb kinase, a pivotal kinase in NFkB activation and TNFa production. Chronic alcohol exposure also increases IKKb kinase activity in macrophages. We hypothesize that chronic alcohol exposure modulates hsp90 and gp96 in macrophages and regulates TLR4 induced NFkB activation and TNFa production, contributing to liver injury. Thus, hsp90 and gp96 play an important role in the pathophysiology of ALD. The Specific Aims are as follows: 1) To determine the activation and function of hsp90 in alcoholic liver injury by: A) Measuring chaperone activity, dimer formation, and acetylation of hsp90 in whole liver and isolated Kupffer cells from alcohol-fed mice. B) Evaluating the effect of hsp90 inhibitors, 17-AAG and/or 17-DMAG in induction of pro-inflammatory cytokine production, and alleviation of alcoholic liver injury. 2) To examine the role of hsp90 in alcohol-induced sensitization to LPS-induced inflammatory cytokine production by: A) Studying the interactions of MyD88- depepndent down-stream signaling molecules, IRAK-1 and IKK with co-chaperone cdc37 and hsp90 in alcoholic Kupffer cells and whole livers. B) Examining whether hsp90 is required in chronic alcohol mediated LPS-induced, MyD88 independent signaling. C) Evaluating the interaction of Gp96, an ER form of hsp90, with TLR4 in alcohol exposed hepatic macrophages/KCs. 3) To assess the mechanisms by which chronic alcohol induces hsp90 in macrophages by: A) Characterization of the binding of HSF-1, NFkB, stimulatory protein-1 (Sp1) and STAT1 by chromatin immunoprecipitation analysis. B) Delineating the effect of HSF-1 deficiency and HSF-1 inhibition using siRNA on induction of hsp90. C) Determining the role of ROS dependent HSF1 activation on induction of hsp90 by evaluating effect of Rac1 and NADPH oxidase in alcohol-exposed macrophages.

. 摘要肝脏驻留巨噬细胞的活化和促炎细胞因子产生的增加是肝硬化的标志。酒精性肝病（ALD）的发病机制。酒精诱导的氧化应激在巨噬细胞活化热休克蛋白是由氧化应激诱导产生的，监护人TLR 4的热休克蛋白90（hsp 90）和Grp 94/gp 96分子伴侣信号分子调节炎症细胞因子的途径。因此，热休克蛋白90和糖蛋白96可能与应激和炎症反应有关。途径，并在ALD的发展中发挥重要作用。我们的初步研究表明长期酗酒与成对喂养的对照相比，在小鼠中喂养增加了分离的Kupffer细胞（KC）中的hsp 90和gp 96。来自慢性酒精暴露巨噬细胞的Hsp 90与NFkB中的关键激酶IKKb激酶相关活化和TNF α产生。慢性酒精暴露也增加IKKb激酶活性，巨噬细胞我们假设慢性酒精暴露调节巨噬细胞中的hsp 90和gp 96，调节TLR 4诱导的NFkB活化和TNF α产生，促进肝损伤。因此，HSP 90和 gp 96在ALD的病理生理过程中起重要作用。具体目的如下：1）确定 hsp 90在酒精性肝损伤中活化和功能：A）测定伴侣活性、二聚体形成，和乙酰化的热休克蛋白90在整个肝脏和分离的库普弗细胞从酒精喂养的小鼠。（B）评估hsp 90抑制剂、17-AAG和/或17-DMAG在诱导促炎细胞因子中的作用产生和减轻酒精性肝损伤。2)探讨热休克蛋白90在酒精诱导的肝细胞凋亡中的作用。 A）研究MyD 88-MyD 88的相互作用，依赖性下游信号分子IRAK-1和IKK与共分子伴侣cdc 37和hsp 90，酒精枯否细胞和整个肝脏B）检查在慢性酒精介导的糖尿病中是否需要hsp 90。 LPS诱导的MyD 88独立信号传导。C）评估Gp 96（一种ER形式的hsp 90）与酒精暴露的肝巨噬细胞/KCs中的TLR 4。3)评估慢性酒精中毒 A）表征HSF-1、NFkB、刺激蛋白-1 (Sp1)染色质免疫沉淀分析STAT 1。B）描述HSF-1缺乏的影响，使用siRNA对hsp 90诱导的HSF-1抑制。C）确定ROS依赖性HSF 1的作用通过评估Rac 1和NADPH氧化酶在酒精暴露中的作用，巨噬细胞