CONTROL OF DRUG AND ETHANOL METABOLISM

描述：(改编自《调查者摘要》)最近 在资助期，我们收集了支持这一假说的证据 酒精性肝损伤与枯否细胞活化有关 肠源性内毒素、氧化剂和有毒细胞因子TNFa。此外，使用 调查人员发现，筑本和弗伦奇的肠道喂养模式 雌性大鼠和人类一样，表现出比雄性更多的伤害，并且有更高的 血液内毒素水平。这一发现导致我们发现雌激素 增加库普弗细胞内毒素受体(CD14)和 产生肿瘤坏死因子(.当肠道消毒时，这两个参数都降低了 使用不可吸收的抗生素。此外，调查人员还表明， Kupffer细胞对急性乙醇引起的内毒素敏感性增加 与CD14表达增加有关。迄今为止，关于机制的报告 早期酒精诱导的肝病依赖于抑制剂的工作，这 可能不是很具体。因此，为了填补我们知识中的关键空白， 研究人员将使用基因敲除技术，在特定受体或蛋白质 基因被删除是为了获得关于因果事件的明确信息 早期酒精性肝损伤的机制。统一的假设我们 将在这里使用基因敲除技术测试内毒素激活库普弗 细胞产生氧化剂，增加TNFa的产生，导致早期 酒精性肝损伤。调查人员还提出，性别 不同之处可以用这个假设来解释。最初，在目标1中，PI 将优化、表征和验证肠内酒精的小鼠模型 通过测定野生型小鼠的病理来分娩，这是背景 所选淘汰赛的菌株。在目标2中，PI将确定主要的 早期酒精性肝病的氧化剂来源是 乙醇诱导的细胞色素P450(细胞色素P450)，研究人员预计将 主要来源于实质细胞或来自Kupffer细胞的NADPH氧化酶。 P47Phox基因敲除小鼠缺乏NADPH氧化酶，将与之进行比较 用CYP2E1基因敲除来检验自由基生成通过 库普弗细胞是早期酒精性肝损伤的主要事件，使用 肠道模型的特点：1.利用内毒素受体(CD14) 肠内研究中的基因敲除和TNFa受体1(TNFR1)基因敲除 酒精，调查人员将测试内毒素和TNFa是 在目标3中与早期酒精性肝损伤有因果关系。在目标4中， 雌激素受体基因敲除小鼠将被用来确定性别差异 在酒精引起的肝脏损伤中可以通过参与 雌激素。这项工作是及时和令人兴奋的，因为它将提供一个新的企业 老鼠的喂养模式，将允许研究人员和其他人 探讨特定蛋白质和酶在酒精性肝损伤中的作用 疾病使用基因敲除技术的力量。这将使我们的定位独一无二 提供明确的新信息并填补我们知识中的关键空白 早期酒精性肝损伤机制的探讨。

DESCRIPTION: (Adapted from the Investigator's Abstract) During the recent funding period, we have collected evidence supporting the hypothesis that early alcohol-induced liver injury involves activation of Kupffer cells by gut-derived endotoxin, oxidants and the toxic cytokine TNFa. Moreover, using the enteral feeding model of Tsukamoto and French, the investigators discovered that female rats, like humans, exhibit more injury than males and have higher blood endotoxin levels. This finding led to our discovery that estrogen increases Kupffer cell expression of the endotoxin receptor (CD14) and production of TNF(. Both parameters were decreased when the gut was sterilized with non-absorbable antibiotics. Moreover the investigators showed that increased sensitivity of Kupffer cells to endotoxin caused by acute ethanol is associated with an increase in CD14 expression. To date, reports on mechanisms of early alcohol-induced liver disease relied on work with inhibitors, which may not be specific. Accordingly, to fill critical gaps in our knowledge, the investigators will use knockout technology where specific receptors or proteins are genetically deleted to obtain clear information on causal events in mechanisms of early alcohol-induced liver injury. The unifying hypothesis we will test here using knockout technology is that endotoxin activates Kupffer cells to produce oxidants which increase TNFa production leading to early alcohol-induced liver injury. The investigators also proposed that gender differences could be explained by this hypothesis. Initially, in Aim 1 the PI will optimize, characterize and validate a mouse model of enteral alcohol delivery by determining pathology in wild-type mice, which are background strains for selected knockouts. In aim 2, the PI will determine if the primary source of oxidants in early alcohol-induced liver disease is the ethanol-inducible cytochrome P450 (CYP2E1) which the investigators expect will come predominantly from parenchymal cells or NADPH oxidase from Kupffer cells. The p47phox knockout mouse, which is NADPH oxidase deficient, will be compared wit h the CYP2E1 knockout to test the hypothesis that radical generation by Kupffer cells is a primary event in early alcohol-induced liver injury, using the enteral model characterized in Aim 1. Using the endotoxin receptor (CD14) knockout and the TNFa receptor 1 (TNFR1) knockout in enteral studies with alcohol, the investigators will test the hypothesis that endotoxin and TNFa are causally involved in early alcohol-induced liver injury in Aim 3. In Aim 4, estrogen receptor knockout mice will be used to determine if gender differences in liver injury due to alcohol can be explained by the involvement of estrogens. This work is timely and exciting since it will provide a new enteral feeding model in the mouse that will allow the investigators and others to investigate the roles of specific proteins and enzymes in alcohol-induced liver disease using the power of knockout technology. This will uniquely position us to provide unequivocal new information and fill critical gaps in our knowledge on mechanisms of early alcohol-induced liver injury.