MOLECULAR PATHOPHYSIOLOGY OF ALCHOLIC LIVER DISEASE

酒精性肝病的分子病理生理学

• 批准号: 3462676
• 负责人: FRANCIS R WEINER
• 金额: $ 11.81万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-08-01 至 1991-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3462676
• 关键词: DNA methylation; alcoholic liver cirrhosis; alcoholism /alcohol abuse; alleles; biological polymorphism; biopsy; corticosteroids; fibrogenesis; genetic library; genetic markers; genetic transcription; histopathology; human subject; liver cells; messenger RNA; molecular cloning; molecular pathology; nucleic acid sequence; sex hormones; tissue /cell culture

The objectives are to obtain a better understanding of the molecular effects of alcohol upon hepatic function as well as the pathogenesis of hepatic fibrosis in alcoholic liver disease. Two general areas of inquiry will be investigated: (1) What levels of gene regulation and specifically collagen gene regulation are affected in alcoholic liver disease? This will involve an analysis of mRNA steady state levels by cell free protein synthesis and RNA-DNA hybridization. Quantitative determinations of changes in several specific mRNA species including types I, III, and IV procollagen, albumin, fibronectin and -actin will be undertaken. Nuclear transcriptional run-off analysis will be performed in order to examine the basis for observed changes in steady state mRNA contents. Collagen gene promoter structure and function will be determined by constructing partial genomic libraries from cirrhotics and non-cirrhotic alcoholics, isolating and purifying the collagen promoter, and analyzing it by means of S1 nuclease sensitivity methylation, DNA sequencing, and use of a transient expression vector (pSV-0-CAT). These latter studies may explain variations in an individual's fibrotic response to alcohol. (2) What factors predispose an individual to develop alcoholic cirrhosis? This will involve a genetic analysis of the collagen gene using DNA polymorphisms and analysis of the role of sex hormones and corticosteroids in hepatic collagen synthesis using cell culture techniques, molecular hybridization studies and in vivo transient expression vector analysis. The results of such experiments will help us expand our knowledge of the molecular mechanisms responsible for hepatic fibrosis in man. This knowledge will help in developing diagnostic methods for evaluating fibrogenesis predicting susceptibility to develop cirrhosis, and provide the basis for better therapy. These studies will also facilitate my long term objective to become an independent investigator in academic hepatology with expertise in current areas of molecular biology.

目的是为了更好地了解分子 酒精对肝功能的影响及其发病机制 酒精性肝病的肝纤维化。调查的两个一般领域 将调查：(1)什么水平的基因调控和具体 胶原蛋白基因调控在酒精性肝病中有影响吗？这 将涉及到通过细胞游离蛋白来分析信使核糖核酸的稳定水平 合成和RNA-DNA杂交。三聚氰胺的定量测定 包括I、III和IV型在内的几个特定mRNA物种的变化 将进行前胶原、白蛋白、纤维连接蛋白和肌动蛋白检查。核子 将进行转录径流分析，以检查 为观察到的稳态mRNA含量的变化奠定了基础。胶原蛋白基因 启动子的结构和功能将通过构建部分 来自肝硬化性和非肝硬化性酒精中毒患者的基因组文库，分离 纯化胶原蛋白启动子，并用S1进行分析 核酸酶敏感度甲基化、DNA测序和使用瞬时 表达载体(PSV-0-CAT)。这些后一项研究可能会解释变异 在个体对酒精的纤维化反应中。(2)什么因素 易患酒精性肝硬变？这将涉及到 胶原蛋白基因的DNA多态和遗传分析 性激素和皮质类固醇在肝脏中的作用分析 利用细胞培养技术、分子杂交技术合成胶原 研究和体内瞬时表达载体分析。结果是 这样的实验将帮助我们扩大对分子的了解 人类肝纤维化的机制。这一知识将会 帮助开发评估纤维化形成的诊断方法 预测发展为肝硬变的易感性，并提供依据 更好的治疗方法。这些研究也将有助于我的长期目标 成为具有专业知识的学术肝病的独立研究员 在当前的分子生物学领域。