INDUCTION OF AP1 AND NFKB BY ETHANOL

描述（由申请人提供）：患者的酒精性肝病似乎从脂肪变性进展为酒精性肝炎、纤维化，最后是肝硬化。最近在啮齿动物中的研究已经阐明了慢性酒精摄入导致脂肪变性和脂肪性肝炎的许多途径。 然而，肝纤维化和肝硬化的进展在很大程度上是未知的。该项目的总体目标是确定脂肪性肝炎导致酒精性肝病纤维化的机制。 我们将使用小鼠胃内灌洗模型进行研究，以利用小鼠遗传学和肝星状细胞的原代培养。这一建议基于几个基本假设：1。肝星状细胞（HSC）的活化是酒精性肝病从脂肪性肝炎进展为纤维化的关键介质。2. HSC在培养物中的活化再现了体内活化和肝纤维化的关键组分。3.脂多糖（LPS）是酒精诱导纤维化的关键因素。4.雌性小鼠比雄性小鼠更容易患酒精性纤维化。 该项目的具体目标是： 1.为了确定NF-κ B 3转录活性是否是HSC活化的早期标志物。 2.探讨酒精性肝病中肝星状细胞的活化和增殖是否促进了1型胶原的表达。 3.确定在酒精性肝病中活化的HSC中纤维形成是否超过基质降解。 实验设计将使用我们实验室创建的三种新型转基因小鼠。转基因系表达由胶原α 1（I）启动子和增强子、平滑肌肌动蛋白启动子或NF-κ B反应元件驱动的报告基因。这些小鼠将通过胃内乙醇喂养模型长期接受乙醇治疗。通过结合使用这些转基因小鼠的体内和培养研究，我们的目标是发现酒精性肝纤维化的分子发病机制的新见解。

DESCRIPTION (provided by applicant): Alcoholic liver disease in patients appears to progress from steatosis to alcoholic hepatitis, fibrosis, and finally cirrhosis. Recent studies in rodents have elucidated many of the pathways by which chronic alcoholic intake leads to steatosis and steatohepatitis. However, the progression to hepatic fibrosis and cirrhosis is largely unknown. The overall goal of this project is to define the mechanism by which steatohepatitis leads to fibrosis in alcoholic liver disease. We will perform studies using the intragastric lavage model in mice to take advantage of mouse genetics and using primary cultures of hepatic stellate cells. This proposal is based on several underlying hypotheses: 1. Activation of the hepatic stellate cell (HSC) is the key mediator of the progression of alcoholic liver disease from steatohepatitis to fibrosis. 2. Activation of HSCs in culture recapitulates critical components of activation and hepatic fibrosis in vivo. 3. Lipopolysaccharide (LPS) is a critical factor in alcohol-induced fibrosis. 4. Female mice are more susceptible to alcoholic fibrosis than male mice. The specific aims to be addressed in this project are: 1. To determine if NF-rJ3 transcriptional activity is an early marker of HSC activation. 2. To determine if HSC activation and proliferation proceeds type 1 collagen expression in alcoholic liver disease. 3. To determine if fibrogenesis exceeds matrix degradation in activated HSCs in alcoholic liver disease. The experimental design will use three novel transgenic mice that were created in our laboratory. The trangenic lines express reporter genes driven by the collagen alpha1(I) promoter and enhancer, the smooth muscle a actin promoter, or an NF-kappaB responsive element. These mice will be treated chronically with ethanol by the intragastric ethanol feeding model. By combining in vivo and in culture studies using these transgenic mice, our goal is to discover new insights into the molecular pathogenesis of alcoholic liver fibrosis.