Acute Ethanol-Induced Innate Immune Response in Liver

描述（由申请方提供）：内毒素参与早期酒精诱导的肝损伤已得到充分证实;支持发病机制涉及先天免疫和炎症介质方面的假设。 许多报道已经清楚地表明，枯否细胞在乙醇的发病机制中起着关键作用。具体而言，研究表明，缺乏内毒素受体CD 14（主要在枯否细胞上表达）的小鼠对慢性酒精诱导的肝损伤具有抗性。 这些数据表明，CD 14信号可能是酒精相关肝损伤的关键组成部分。 总体假设是来自肠道的LPS激活枯否细胞，导致氧化剂产生增加和随后的TNF-α释放。 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。 尽管关于内毒素在乙醇诱导的肝损伤中的作用有大量的新信息，但我们的理解仍然存在重大差距。例如，参与LPS诱导的氧化剂产生的信号传导机制、发病机制中CD 14和相关信号传导组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基本假设：1. PI 3激酶介导LPS诱导的枯否细胞中NADPH氧化酶产生超氧化物。 2.急性乙醇给药后氧化敏感性转录因子NF-κ B和AP-1调节CD 14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。 下面的目标将使用基因递送技术和敲除小鼠技术来解决这些关键问题，这些关键问题与LPS，PI 3激酶激活和CD 14表达在急性乙醇和慢性乙醇毒性中的作用有关。

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.