BASES OF CIRRHOSIS IN ALCOHOLIC LIVER DISEASE

酒精性肝病中肝硬化的基础

• 批准号: 2855741
• 负责人: MARK ALLEN ZERN
• 金额: $ 16.27万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-09-01 至 1999-08-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2855741
• 关键词: BASES; CIRRHOSIS; ALCOHOLIC; LIVER; DISEASE

This proposal represents a continuation and expansion of the PI's initial objective of attempting to understand the molecular mechanisms responsible for hepatic fibrogenesis. One difficulty in their search for answers has been the lack of an adequate rodent model for the development of hepatic injury and fibrosis caused by ethanol. Thus, they propose to develop improved in vitro and in vivo methods of ethanol-induced ad liver injury and to employ the models to investigate the significance of two interrelated factors, tissue transglutaminase (tTG) and nuclear factor-kappa B (NF-kappaB), in the process of hepatic injury and fibrosis. Specific Aims: 1) to develop new models of ethanol-induced hepatic injury and fibrogenesis; 2) to delineate the role of NF-kappaB as a mediator of hepatic injury and fibrogenesis; 3) to assess how tTG affects hepatic injury and fibrogenesis; and, 4) to determine the relationship between the GTP-binding and cross-linking activities of tTG. Methods: Isolated calls from Osteogenic Disorder Shionogi (ODS) rats will be treated with ethanol to characterize aspects of the injury and activation process. The mechanisms by which ethanol injures the hepatocytes will be evaluated by malondialdehyde levels, Northern blot hybridization analysis of cytokines and tTG, and gel retardation assays of NF-kappaB. ODS rats will be fed the Lieber- DeCarli diet in an attempt to develop an improved model for chronic alcoholic liver disease. Once the models are established, attempts will be made to inhibit injury in the model systems with novel therapeutic agents. Attempt to determine pathophysiologic significance of NF-kappaB in the injury process will be under undertaken using a proteasome inhibitor which blocks the degradation of IkappaB. Mutagenesis directed to the NF-kappaB motif as well as analysis of other regulatory regions of the tTG promoter will be used to determine the expression a tTG in in vivo injury models. Specific a1-adrenergic receptor agonist and antagonist treatment of hepatocytes will be undertaken to investigate the interaction between the GTPase and cross-linking activity of tTG. Health Relatedness: It is hoped that by better understanding the molecular bases of alcoholic liver diseases, effective and rational therapy can be developed.

该提案代表了PI的延续和扩展， 最初的目标是试图了解分子机制 导致肝纤维化 他们搜寻的一个困难是 答案是缺乏足够的啮齿动物模型， 乙醇引起的肝损伤和纤维化的发展。 因此，在本发明中， 他们提出开发改进的体外和体内方法， 乙醇诱导的AD肝损伤，并利用该模型研究 两个相互关联的因素，组织转氨酶， (tTG)和核因子-κ B（NF-κ B）在肝硬化过程中的作用 损伤和纤维化。 具体目标：1）开发新的模型 乙醇诱导的肝损伤和肝纤维化; 2）描述 NF-κ B作为肝损伤和纤维化发生介质的作用; 3） 评估tTG如何影响肝损伤和纤维化;以及，4） 确定GTP结合和交联之间的关系 tTG的活动。 方法：从成骨障碍中分离呼叫 将用乙醇处理Shionogi（ODS）大鼠，以表征 损伤和激活过程。 乙醇的作用机制 肝细胞损伤将通过丙二醛水平进行评估， 细胞因子和tTG的北方印迹杂交分析，以及凝胶电泳 NF-κ B的阻滞测定。 混乱办的老鼠会被喂利伯- DeCarli饮食，试图开发一种改进的慢性 酒精性肝病 一旦建立了模型， 在模型系统中用新的治疗剂抑制损伤 剂. 尝试确定NF-κ B的病理生理意义 在损伤过程中的作用将通过蛋白酶体进行 抑制剂，其阻断IkappaB的降解。 定向诱变 NF-κ B基序以及其他调控区域的分析 tTG启动子的表达将用于确定tTG在 体内损伤模型。 特异性α 1-肾上腺素能受体激动剂和 将进行肝细胞拮抗剂治疗，以研究 GTG与tTG交联活性的相互作用。 健康相关性：希望通过更好地了解 酒精性肝病的分子基础，有效性和合理性 可以开发治疗。