BASES OF CIRRHOSIS IN ALCOHOLIC LIVER DISEASE

这项提议是对国际和平倡议的延续和扩展 最初的目标是试图了解分子机制 对肝纤维化负有责任。他们寻找过程中的一个困难 答案是缺乏足够的啮齿动物模型来应对 酒精性肝损伤和肝纤维化的进展。因此， 他们建议开发改进的体外和体内方法 酒精性肝损伤模型的建立及应用 组织转谷氨酰胺酶两个相互关联的因素的意义 (TTG)和核因子-kappaB(NF-kappaB)在肝纤维化过程中的作用 损伤和纤维化。具体目标：1)开发新的模式 酒精性肝损伤和肝纤维化；2)勾画 核因子-kappaB在肝损伤和肝纤维化中的作用 评估tTG如何影响肝损伤和纤维化；以及，4) 确定GTP结合和交联剂之间的关系 TTG活性。方法：骨源性疾病患者的孤立叫声 Shionogi(Ods)大鼠将用乙醇处理以表征各方面 关于损伤和激活过程的信息。乙醇的作用机制 对肝细胞的损伤将通过丙二醛水平来评估， 细胞因子与tTG、凝胶的Northern印迹杂交分析 核因子-kappaB的滞留分析。消耗臭氧层物质的大鼠将被喂食利伯- DeCarli节食，试图开发一种改进的慢性 酒精性肝病。一旦建立了模型，尝试将 用新的治疗方法在模型系统中抑制损伤 探员们。核因子-kappaB的病理生理意义 在损伤过程中将使用蛋白酶体进行 阻断IkappaB降解的抑制剂。定向诱变 对核因子-kappaB基序以及其他调控区域的分析 将使用tTG启动子来确定αtTG在 活体损伤模型。特异性A1受体激动剂和 肝细胞的拮抗剂治疗将进行调查 GTP酶与tTG交联度之间的相互作用。 与健康相关：希望通过更好地了解 酒精性肝病的分子基础，有效而合理 治疗方法是可以开发的。

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.