BASES OF CIRRHOSIS IN ALCOHOLIC LIVER DISEASE

酒精性肝病中肝硬化的基础

• 批准号: 2043448
• 负责人: MARK ALLEN ZERN
• 金额: $ 29.76万
• 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 1996-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2043448
• 关键词: acetaldehyde; alcoholic liver cirrhosis; antisense nucleic acid; cell type; collagen; colony stimulating factor; cytokine; electron microscopy; enzyme linked immunosorbent assay; ethanol; extracellular matrix; fibrogenesis; gene expression; genetic transcription; growth factor receptors; human subject; in situ hybridization; interleukin 1; laboratory rat; liver cells; messenger RNA; northern blottings; nuclear runoff assay; nucleic acid probes; pathogenic diet; prostaglandin E; protein biosynthesis; radioimmunoassay; tissue /cell culture; transforming growth factors; tumor necrosis factor alpha

This proposal represents a continuation and expansion of our initial objective of attempting to understand the molecular mechanisms responsible for hepatic fibrogenesis. The final common pathway which occurs in all forms of liver fibrosis regardless of etiology is the increased deposition of extracellular matrix. We hypothesize that this process, which includes cell damage, recruitment and proliferation of inflammatory and mesenchymal cells, as well as extracellular matrix synthesis and degradation, is orchestrated by a variety of soluble peptides including cytokines. The aims of the present proposal are to define the role of these modulatory cytokines in hepatic fibrosis and to investigate the mechanisms by which these effector proteins can be regulated. Specific Aims: 1. To delineate the temporal expression of the cytokines involved in the pathogenesis of alcoholic liver disease. 2. To determine which cell types are synthesizing cytokines in alcoholic liver disease. 3. To ascertain how ethanol influences cytokine gene expression. 4. To determine the effects of prostaglandin E2 (PGE2) on matrix deposition, cytokine synthesis and transforming growth factor-beta (TGF-beta)receptors. 5. To investigate the effects of inhibiting cytokine activity in our rat model of alcoholic liver disease. Methods: The effects of the fibrogenic cytokines on extracellular matrix protein synthesis and deposition will be studied in primary cultures of hepatocytes, Ito cells, and Kupffer cells, and in the rat intragastric feeding model of alcohol-induced liver fibrosis. Cytokine and extracellular matrix synthesis will be evaluated by Northern hybridization analysis, nuclear run-on assays, morphological analysis, biochemical determinations, and RIAs. In situ hybridization analysis will be employed to localize cytokines in the animal model of alcoholic liver disease and in human liver specimens. The significance of PGE2 as an antifibrogenic agent will be evaluated in the same in vivo and in vitro model systems, while its effects on TGF-beta receptors will be investigated by affinity cross-linking studies. TGF-beta and interleukin-I activity will be inhibited in the in vivo model by the use of specific receptor antagonists. Health Relatedness: By accomplishing the aims of this proposal we shall formulate a general model for the pathogenesis of alcoholic liver disease and thereby aid in the formulation of rational therapy.

这一建议是我们最初建议的延续和扩大。 目的是试图了解分子机制 导致肝纤维化 最后的共同途径 其发生在所有形式的肝纤维化中，无论病因如何 是细胞外基质沉积增加。 我们假设 这个过程，包括细胞损伤，招募和 炎症和间充质细胞的增殖，以及 细胞外基质的合成和降解，是由 包括细胞因子在内的多种可溶性肽。 的目标 目前的建议是确定这些调节的作用， 细胞因子在肝纤维化中的作用，并探讨其机制。 这些效应蛋白可以被调节。具体目标：1。 为了描述参与的细胞因子的时间表达， 酒精性肝病的发病机制。2.以确定哪些 细胞类型在酒精性肝病中合成细胞因子。 3.确定乙醇如何影响细胞因子基因表达。4. 为了确定前列腺素E2（PGE 2）对基质的影响， 沉积、细胞因子合成和转化生长因子-β （TGF-β）受体。5.为了研究抑制 在我们的酒精性肝病大鼠模型中的细胞因子活性。 方法：应用致纤维化细胞因子， 基质蛋白质的合成和沉积将在初级研究 肝细胞、Ito细胞和Kupffer细胞的培养物中， 大鼠灌胃酒精性肝纤维化模型。 细胞因子和细胞外基质合成将通过以下方法进行评价： 北方杂交分析，核连续分析， 形态分析、生化测定和RIA。 在 将采用原位杂交分析来定位细胞因子 在酒精性肝病动物模型和人类肝脏中 标本 PGE 2作为抗纤维化剂的意义将 在相同的体内和体外模型系统中进行评价， 其对TGF-β受体的作用将通过亲和性来研究。 交叉研究。 TGF-β和白细胞介素-I的活性将 在体内模型中通过使用特异性受体 对手。健康相关性：通过实现这一目标， 建议我们将制定一个发病机制的一般模型， 酒精性肝病，从而有助于制定 理性治疗