Acute Ethanol-Induced Innate Immune Response in Liver

急性乙醇诱导的肝脏先天免疫反应

• 批准号: 6878118
• 负责人: MICHAEL D WHEELER
• 金额: $ 25.46万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6878118
• 关键词: AP1 protein; CD14 molecule; Kupffer' s cell; NAD(P)H dehydrogenase; RNase protection assay; ethanol; gel mobility shift assay; gender difference; gene delivery system; genetically modified animals; immunocytochemistry; immunopharmacology; immunoprecipitation; laboratory mouse; lipopolysaccharides; liver disorder; nuclear factor kappa beta; phosphatidylinositol 3 kinase; superoxides; tissue /cell culture; tumor necrosis factor alpha; western blottings

DESCRIPTION (provided by applicant): The involvement of endotoxin in early alcohol-induced liver injury is well established; supporting the hypothesis, that pathogenesis involves aspects of innate immunity and inflammatory mediators. A number of reports have clearly demonstrated that Kupffer cells play a critical role in the pathogenesis due to ethanol. Specifically, it was shown that mice deficient in the endotoxin receptor CD14, which is primarily expressed on Kupffer cells, were resistant to chronic alcohol-induced liver injury. These data suggest that CD14 signaling may be a critical component in alcohol-related liver injury. The overall hypothesis is that LPS from the gut activates Kupffer cells causing an increase in oxidant production and subsequent TNF-alpha release. This hypothesis is strongly supported by a number of studies using gene therapy, knockout and transgenic animals, as well as an in vivo mouse intragastric ethanol-feeding model. Despite a great amount of new information regarding the role of endotoxin in ethanol-induced liver injury, critical gaps still exist in our understanding. For example, the signaling mechanisms involved in LPS- induced oxidant production, the regulation of CD14 and related signaling components in pathogenesis, and the molecular mechanisms determining gender- related differences in injury remain unknown. The purpose of this application is to address the following underlying hypotheses: 1. PI3 kinase mediates LPS-induced NADPH oxidase generation of superoxide in Kupffer cells. 2. Oxidant-sensitive transcription factors NF-kappaB and AP-1 regulate CD14 expression following acute ethanol administration. 3. Gender differences in regulation of innate immune response and transcription factor activation are key to increased susceptibility to ethanol-induced pathogenesis in females. The aims below will use gene delivery techniques and knockout mouse technology to address these critical questions related to roles of LPS, PI3 kinase activation, and CD14 expression in both acute ethanol and chronic ethanol toxicity.

描述（由申请人提供）：内毒素参与早期酒精性肝损伤已得到充分证实；支持这一假设，即发病机制涉及先天免疫和炎症介质的各个方面。 许多报告已经清楚地证明库普弗细胞在乙醇引起的发病机制中发挥着关键作用。具体来说，研究表明，缺乏内毒素受体 CD14（主要在库普弗细胞上表达）的小鼠能够抵抗慢性酒精引起的肝损伤。 这些数据表明 CD14 信号传导可能是酒精相关肝损伤的关键组成部分。 总体假设是，肠道中的 LPS 会激活库普弗细胞，导致氧化剂产生增加，并随后释放 TNF-α。 这一假设得到了许多使用基因治疗、基因敲除和转基因动物以及体内小鼠胃内乙醇喂养模型的研究的有力支持。 尽管有大量关于内毒素在乙醇引起的肝损伤中的作用的新信息，但我们的理解仍然存在严重差距。例如，LPS诱导的氧化剂产生所涉及的信号传导机制、发病机制中CD14和相关信号传导成分的调节以及决定损伤中性别相关差异的分子机制仍然未知。 本申请的目的是解决以下潜在假设： 1. PI3 激酶介导 Kupffer 细胞中 LPS 诱导的 NADPH 氧化酶产生超氧化物。 2.急性乙醇施用后，氧化剂敏感转录因子NF-kappaB和AP-1调节CD14表达。 3.先天免疫反应和转录因子激活调节的性别差异是女性对乙醇诱导的发病机制易感性增加的关键。 以下目标将使用基因递送技术和敲除小鼠技术来解决与 LPS、PI3 激酶激活和 CD14 表达在急性乙醇和慢性乙醇毒性中的作用相关的关键问题。