IN VITRO MODEL OF ETHANOL-DEPENDENT LIVER CELL INJURY

乙醇依赖性肝细胞损伤的体外模型

• 批准号: 3112923
• 负责人: ROBERT G LAMB
• 金额: $ 7.6万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3112923
• 关键词: SDS polyacrylamide gel electrophoresis; alcoholic liver cirrhosis; antibody; antioxidants; autoradiography; cell death; dietary constituent; disease /disorder model; enzyme linked immunosorbent assay; ethanol; glutathione; hepatotoxin; hydrolysis; immunoprecipitation; laboratory rat; linoleate; lipid biosynthesis; lipid peroxides; liver cells; membrane lipids; nucleotidyltransferase; nutrition related tag; pathologic process; peroxidation; phosphatidylcholines; phospholipase A2; phospholipase C; phospholipids; protein purification; tissue /cell culture; tocopherols

Alcohol-dependent liver disease (ALD) is a major public health problem since 75% of all medical deaths associated with alcoholism are attributable to cirrhosis. Unfortunately, the effective treatment and prevention and ALD is limited since the mechanisms by which alcohol induces liver cell necrosis are unresolved. Resolution of this issue would be facilitated by the development of an in vitro liver cell model that mimics the hepatocyte's response to ethanol in vivo such as steatosis and necrosis. Therefore, the primary objectives of these studies are to demonstrate that: 1) primary cultures of adult rat hepatocytes incubated (1 to 14 days) with ethanol (10 to 100mM) are an appropriate model in vitro for determining the mechanisms by which ethanol induces liver cell necrosis; 2) ethanol metabolism creates an "oxidative stress" in liver cells that results in membrane injury when membrane phosphatidylcholines (PC) are peroxidized and then hydrolyzed by phospholipases (PC depletion); 3) ethanol-dependent membrane injury (PC depletion) is reversible, or irreversible, depending on whether membrane PC biosynthesis is, or not, correspondingly increased. The validity of these concepts will be established by incubating cultured hepatocytes with ethanol and agents and increased (linoleate) or decrease (4-methylpyrazole and Vitamin E) ethanol-dependent liver cell injury in vivo and determining the agent mediated changes in ethanol-dependent liver cell injury and the peroxidation, hydrolysis and biosynthesis of membrane PC. Isocaloric (ethanol or carbohydrate) liquid diets will also be acutely and chronically administered to rate to demonstrate that a alcohol-dependent changes in liver cell functions (viability and the peroxidation, hydrolysis and biosynthesis of membrane PC) in cultured hepatocytes are consistent with those observed in the intact animal. In summary, these studies will demonstrate that the peroxidation, hydrolysis and biosynthesis of membrane PC are critical events in ethanol- dependent cell injury. Determining how ethanol and/or other agents such as linoleate and Vitamin E alter membrane PC peroxidation, hydrolysis and biosynthesis will contribute to a better understanding of how to prevent and treat ethanol-induced liver cell injury.

酒精依赖性肝病（ALD）是一个主要的公共卫生问题 由于与酒精中毒相关的所有医疗死亡中有75％是可归因于 肝硬化。不幸的是，有效的治疗和预防和ALD 由于酒精诱导肝细胞的机制而受到限制 坏死尚未解决。解决这个问题将由 模拟的体外肝细胞模型的发展 肝细胞对体内乙醇的反应，例如脂肪变性和坏死。 因此，这些研究的主要目标是证明： 1）成年大鼠肝细胞的原发性培养（1至14天）与 乙醇（10至100mm）是一个合适的体外模型，用于确定 乙醇诱导肝细胞坏死的机制； 2）乙醇代谢在肝细胞中产生“氧化应激” 当膜磷脂酰胆碱（PC）为 过氧化，然后通过磷脂酶（PC耗竭）水解； 3）依赖乙醇的膜损伤（PC耗竭）是可逆的，或 不可逆的，具体取决于膜PC生物合成是否是， 相应增加。 这些概念的有效性将通过孵化培养 乙醇和剂的肝细胞，并增加（linoleate）或减少 （4-甲基吡唑和维生素E）依赖乙醇的肝细胞损伤 体内并确定剂介导的乙醇依赖性肝脏的变化 细胞损伤以及膜的过氧化，水解和生物合成 个人电脑。 等生成（乙醇或碳水化合物）液体饮食也将是敏锐的，并且 长期施用的速率以证明酒精依赖性 肝细胞功能的变化（活力和过氧化，水解 培养的肝细胞中膜PC的生物合成是一致的 与完整动物中观察到的那些。 总而言之，这些研究将证明过氧化， 膜PC的水解和生物合成是乙醇的关键事件 依赖的细胞损伤。确定乙醇和/或其他代理（例如 Linoleate和维生素E改变了膜PC的过氧化，水解和 生物合成将有助于更好地理解如何预防 并治疗乙醇诱导的肝细胞损伤。