LIPID ALDEHYDES AND ETHANOL INDUCED LIVER DAMAGE

脂醛和乙醇引起的肝损伤

• 批准号: 2894040
• 负责人: DENNIS PETERSEN
• 金额: $ 20.43万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-07-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2894040
• 关键词: adduct; alcoholism /alcohol abuse; aldehydes; autoantibody; ethanol; free radicals; genetic regulation; glutathione; histology; immunocytochemistry; immunotoxicity; laboratory rat; lipid peroxides; liver metabolism; liver toxic disorder; molecular cloning; molecular pathology; peroxidation; protein sequence; toxicology

The aim of this project is to investigate the ability of chronic ethanol ingestion to stimulate hepatic production of chemically reactive aldehydic products having the potential to function as intermediates in alcohol- induced liver damage. The proposed studies are designed to identify correlations of alcohol-induced changes in lobular histology and immunocytochemistry with biochemical and molecular alterations at the level of zonal-specific hepatocytes and isolated mitochondria. Specifically, experiments are described to measure concentrations of "free" malondialdehyde, 4-hydroxynonenal and hexanal in livers of rats following chronic ethanol feeding. Lipid- and protein-complexed carbonyls will also be measured to quantitate aldehydic functions hound to hepatocellular constituents. The mechanisms of production of these aldehydic products will be examined in isolated periportal and perivenous hepatocytes and isolated mitochondria obtained from rats chronically ingesting ethanol. The ability of ethanol and acetaldehyde metabolism to stimulate lipid peroxidation will be evaluated in zonal specific hepatocytes and isolated mitochondria. Steady-state concentrations of malondialdehyde, 4-hydroxynonenal and hexanal will be quantitated as indices of lipid peroxidation in hepatocytes and mitochondrial for determination of correlations between the magnitude of lipid peroxidation, alterations in glutathione concentrations and disruption of ethanol and acetaldehyde oxidation. The ability of aldehydic lipids to interfere with mitochondrial acetaldehyde oxidation will be evaluated in order to test the hypothesis that these biogenic aldehydes inhibit aldehyde dehydrogenase mediated oxidation of acetaldehyde. A final series of experiments are proposed to evaluate the hypothesis that specific proteins in perivenous or periportal hepatocytes are targets for adduct formation with reactive aldehydic products of lipid peroxidation. This will be determined by using antibodies against aldehydelysine and cysteine adducts for immunocytochemical identification of lobular, cellular and subcellular proteins predisposed for formation of aldehyde adducts. Collectively, the design and statistical analyses employed in the proposed experiments will allow identification of significant changes in lobular histochemistry and immunocytochemical parameters causally related to biochemical and molecular changes at the hepatocellular level. The long-term goal of this proposed research is to establish the role of aldehydic products of lipid peroxidation and their adducts in alcohol-induced liver injury and determine the utility of their measurement as an early indicator of hepatocellular liver damage.

该项目的目的是研究慢性乙醇的能力 摄入刺激肝脏产生化学反应性醛 有潜力作为酒精中间体的产品 诱发肝损伤。拟议的研究旨在确定 酒精引起的小叶组织学变化与 免疫细胞化学与生化和分子改变 区域特异性肝细胞和分离线粒体的水平。 具体来说，实验被描述为测量浓度 大鼠肝脏中“游离”丙二醛、4-羟基壬烯醛和己醛 长期乙醇喂养后。脂质和蛋白质复合羰基 还将测量以定量醛类功能 肝细胞成分。这些物质的产生机制 醛类产品将在隔离的门静脉周围和静脉周围进行检查 长期从大鼠身上获得的肝细胞和分离的线粒体 摄入乙醇。乙醇和乙醛的代谢能力 刺激脂质过氧化将在区域特定中进行评估 肝细胞和分离的线粒体。 稳态浓度 丙二醛、4-羟基壬烯醛和己醛的定量为 肝细胞和线粒体脂质过氧化指数 确定脂质过氧化程度之间的相关性， 谷胱甘肽浓度的改变和乙醇的破坏 乙醛氧化。醛类脂质干扰的能力 将评估线粒体乙醛氧化以测试 这些生物醛抑制醛的假设 脱氢酶介导的乙醛氧化。 最后一个系列 提出实验来评估特定蛋白质的假设 静脉周围或门静脉周围肝细胞是加合物形成的靶标 与脂质过氧化的反应性醛产物反应。这将是 使用抗醛赖氨酸和半胱氨酸加合物的抗体测定 用于小叶、细胞和亚细胞的免疫细胞化学鉴定 蛋白质易于形成醛加合物。总的来说， 拟议实验中采用的设计和统计分析将 允许识别小叶组织化学的显着变化和 免疫细胞化学参数与生化和因果关系 肝细胞水平的分子变化。本次活动的长远目标 拟议的研究是确定醛类产物对脂质的作用 酒精性肝损伤中的过氧化及其加合物 确定其测量作为早期指标的效用 肝细胞肝损伤。