Acute alcohol and calcium-dependent LPS-triggered activation of macrophages

急性酒精和钙依赖性脂多糖触发巨噬细胞激活

• 批准号: 8462176
• 负责人: Angela Dolganiuc
• 金额: $ 33.11万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8462176
• 关键词: Acute; Address; Alcohol abuse; Alcohol consumption; Alcohols; Binding; Calcineurin; Calcium; Calmodulin; Cardiovascular system; Cells; Cellular translocation; Chelating Agents; Chemicals; Clinical; Complex; Consumption; Cytoplasm; Data; Doctor of Philosophy; Endoplasmic Reticulum; Ethanol; Event; Exhibits; Goals; Health; Homeostasis; Human; Human body; Hydrochloride Salt; Immune; Immune system; Immunity; Impairment; In Vitro; Infection; Inflammatory; Inositol; Investigation; Ion Channel; Kupffer Cells; Lead; Lipopolysaccharides; Liver; Macrophage Activation; Mediating; Mitochondria; Modality; Modeling; Molecular; Mus; NFAT Pathway; Nuclear; Pathway interactions; Peritoneal; Peritoneal Macrophages; Phosphorylation; Phosphotransferases; Play; Potassium; Potassium Channel; Principal Investigator; Probability; Production; Regulation; Reporting; Research Proposals; Role; Septic Shock; Signal Transduction; System; TRAF6 gene; Therapeutic; Tissues; Trauma; Tumor Necrosis Factor Activation; Tumor Necrosis Factor-alpha; Wild Type Mouse; alcohol exposure; base; cell type; cytokine; human IRAK1 protein; iberiotoxin; immune activation; improved; in vitro Model; in vivo; inhibitor/antagonist; large-conductance calcium-activated potassium channels; macrophage; monocyte; novel; nuclear factors of activated T-cells; patch clamp; pathogen; paxillin; programs; receptor; toll-like receptor 4; voltage

DESCRIPTION (provided by applicant): Alcohol (ethanol, EtOH) consumption exhibits a wide variety of effect on the human body, ranging from beneficial effect on the cardiovascular system to detrimental effects on other systems, including suppression of the immune system. The mechanisms leading to such versatile effect of EtOH are largely unknown. The goal of this proposal is to define the influence of EtOH on molecular mechanisms involved in regulation of Ca2+-dependent LPS-triggered macrophage (Mf) activation. Our preliminary results suggest that EtOH inhibits pathogen-mediated Mf activation, specifically, upon acute EtOH exposure Mf produce less pro-inflammatory cytokine TNF? when a bacterial-derived product, lipopolysaccharide (LPS) triggers cell activation via Toll-like receptor 4 (TLR4). We provide novel preliminary data that engagement of TLR4 with LPS triggers rapid increase in cytoplasmic Ca2+ in Mf. Moreover, Ca2+ depletion or chemical block of Ca2+ influx abolished LPS-induced TNF? production in Mf. EtOH impairs LPS-induced phosphorylation of CaMKII, a Ca2+-dependent kinase with key role in assembly of initial TLR4 signaling complex. EtOH stimulates Ca2+-dependent activation of NF-AT, a nuclear factor that regulates TNF? production. Based on our preliminary data, we postulated that Ca2+ plays a key role in TLR4-mediated Mf activation and second, EtOH-induced impairment of Ca2+-mediated signaling in Mf. We hypothesize that a) EtOH- conditioned modulation of Ca2+ in the cytoplasm leads to defective signaling via initial TLR4 signaling complex, and/or b) alcohol modulates the previously unknown/under-estimated LPS-triggered Ca2+-calmodulin-calcineurin-CaMKII-NF-AT pathway. We hypothesized that the mechanisms of the acute EtOH-conditioned impairment of LPS-induced Mf activation include alterations of Ca2+ homeostasis and induce modulation of large conductance Ca2+ activated potassium channels. This proposal addresses our hypothesis to unravel the role of Ca2+ in EtOH-induced impairment of TLR4-mediated innate immune activation. PUBLIC HEALTH RELEVANCE: Alcohol consumption exhibits a wide variety of effect on human body, ranging from beneficial effect on cardiovascular system to detrimental effects on all other system, including suppression of the immune system. The mechanisms leading to such versatile effect of EtOH are largely unknown. The overall goal of this research proposal is to define the influence of alcohol on the molecular mechanisms involved in regulation macrophage activation. Macrophages (Mf) produce pro-inflammatory cytokine TNF? when a bacterial- derived product, lipopolysaccharide (LPS) binds to the Toll-like receptor 4 (TLR4). Alcohol inhibits LPS-induced TNF? production. We provide novel preliminary data that engagement of TLR4 increases the cytoplasmic Ca2+ in Mf. More importantly and similar to acute alcohol, Ca2+ depletion or chemical block abolished the LPS-induced TNF? production. Here we postulated that, first; Ca2+ plays a key role in TLR4-mediated Mf activation and second, alcohol modulates the Ca2+ homeostasis in Mf. We hypothesize that a) alcohol-conditioned modulation of Ca2+ leads to defective formation of the initial TLR4 signaling complex, and/or b) alcohol modulates the previously unknown/under-estimated LPS-triggered Ca2+-calmodulin-calcineurin-CaMKII-NFAT pathway. We further hypothesized that acute alcohol modulates the Ca2+-dependent large conductance Ca2+ activated potassium channels. This proposal will address our hypothesis in order to unravel the role of Ca2+ in EtOH-induced impairment of TLR4-mediated innate immune activation.

描述（由申请人提供）：酒精（乙醇，ETOH）的消费对人体表现出各种影响，从对心血管系统的有益作用到对其他系统的有害影响，包括对免疫系统的抑制。导致ETOH多功能效应的机制在很大程度上未知。该建议的目的是定义EtOH对Ca2+依赖性LPS触发的巨噬细胞（MF）激活的调节的分子机制的影响。我们的初步结果表明，ETOH抑制病原体介导的MF激活，特别是在急性ETOH暴露MF时，MF产生的促炎性细胞因子TNF较少？当细菌衍生的产物时，脂多糖（LPS）通过Toll样受体4（TLR4）触发细胞激活。我们提供了新的初步数据，即TLR4与LPS触发的MF中的细胞质Ca2+迅速增加。此外，Ca2+消耗或Ca2+流入的化学块废除了LPS诱导的TNF？ MF生产。 ETOH损害了LPS诱导的CAMKII的磷酸化，CAMKII是一种Ca2+依赖性激酶，在初始TLR4信号复合物组装中具有关键作用。 ETOH刺激了Ca2+依赖性的NF-AT激活，NF-AT是调节TNF的核因子？生产。基于我们的初步数据，我们假设Ca2+在TLR4介导的MF激活中起关键作用，其次是ETOH诱导的MF中Ca2+介导的信号传导的损伤。我们假设a）在细胞质中Ca2+的eTOH条件调节导致通过初始TLR4信号传导复合物和/或b）醇调节先前未知/未知的LPS触发的CA2+-calmodulin-calmodulin-calcinerin-camkii-camkii-Camkii-nf-at途径的信号传导。我们假设LPS诱导的MF激活的急性EtOH条件损伤的机制包括改变Ca2+稳态的改变以及诱导大型电导率Ca2+活化钾通道的调节。该提议介绍了我们的假设，以阐明Ca2+在ETOH诱导的TLR4介导的先天免疫激活中的作用。公共卫生相关性：饮酒对人体具有多种影响，从对心血管系统的有益影响到对所有其他系统的有害影响，包括抑制免疫系统。导致ETOH多功能效应的机制在很大程度上未知。该研究建议的总体目标是定义酒精对调节巨噬细胞激活所涉及的分子机制的影响。巨噬细胞（MF）产生促炎性细胞因子TNF？当细菌衍生产物时，脂多糖（LPS）与Toll样受体4（TLR4）结合。酒精会抑制LPS诱导的TNF？生产。我们提供了新的初步数据，即TLR4的参与增加了MF中的细胞质Ca2+。更重要的是，类似于急性酒精，Ca2+耗竭或化学块废除了LPS诱导的TNF？生产。在这里，我们假设，首先； Ca2+在TLR4介导的MF激活中起关键作用，其次，酒精调节MF中的Ca2+稳态。我们假设a）a）Ca2+的酒精条件调节导致初始TLR4信号传导复合物的形成有缺陷，/或b）酒精调节先前未知/未知的LPS触发的Ca2+ - 钙粘蛋白 - calmodulin-calcineurin-calcineurin-camkii-camkii-nfat pathway。我们进一步假设急性酒精调节Ca2+依赖性的大电导Ca2+活化的钾通道。该建议将解决我们的假设，以揭示Ca2+在ETOH诱导的TLR4介导的先天免疫激活中的作用。