CHRONIC ALCOHOLISM & LIVER ENERGY & LIPID METABOLISM

我工作的长期目标是在生物化学领域 降低慢性酒精所致的肝脏相关病理 消费。主要重点将是对这些人的调查 细胞能量代谢过程中发生的生化变化 酒精性肝病的早期(脂肪肝)。该项目 本建议书中描述的内容旨在1)确定 慢性酒精摄入改变ATP合成系统，2) 提供有关乙醇引起损害的机制的信息 线粒体，以及3)确定这种损伤对细胞的影响 能量代谢。 我们实验室的研究表明，在所有的 乙醇引起的肝线粒体改变：1)肝线粒体功能降低 ATP合成酶复合体的活性，2)降低了能量守恒 电子传递链的NADH-泛醌还原酶片段，以及 3)细胞器的脆性增加，这很可能是 线粒体总补体导致的ATP合成减少 脂肪肝。因此，我们选择了这三个病灶进行进一步的 调查。与酒精有关的心脏改变 ATP合成酶的多肽-磷脂结构将是 下定决心。乙醇诱导的细胞能力的变化 NADH-泛素还原酶复合体驱动AN的形成 将对电化学质子梯度进行更详细的表征。 实验也被设计来确定增加的 酒精喂养大鼠肝脏线粒体的脆性。其中包括 测定肝线粒体周转率。 以上内容将与与酒精相关的调查相结合 肝脏能量代谢的改变与肝细胞和 原位肝脏。肝脏能量状态下降的程度 慢性酒精摄入将在完整肝脏中进行原位研究 在肝细胞中，利用31P核磁共振谱来确定细胞 磷酸化潜力。乙醇诱导的大鼠脑内神经细胞功能改变 体内的线粒体将通过质子动的测量来评估 由肝细胞内的线粒体产生的力。

The long-term objective of my work is to characterize at the biochemical level the liver-associated pathology which results from chronic ethanol consumption. The major emphasis will be an investigation of those biochemical changes that occur in cellular energy metabolism during the early (fatty liver) stage of ethanol-induced liver disease. The project described in this proposal is designed to 1) determine how much the ATP-synthesizing system is altered by chronic ethanol consumption, 2) provide information on the mechanism by which ethanol elicits damage to mitochondria, and 3) determine the effects of such damage on cellular energy metabolism. Studies in our laboratory ot date indicate that among all the ethanol-related alterations in hepatic mitochondria it is 1) the lowered activity of the ATP synthetase complex, 2) decreased energy conservation in the NADH-ubiquinone reductase segment of the electron transport chain, and 3) increased fragility of the organelle that are most likely responsible for decreased ATP synthesis by the total mitochondrial complement in the fatty liver. We have thus selected these three lesions for further investigations. Ethanol-related alterations in the polypeptide-phospholipid structure of the ATP synthetase will be determined. The ethanol-elicited changes in the ability of the NADH-ubiquionone reductase complex to drive the formation of an electrochemical proton gradient will be characterized in further detail. Experiments are also designed to determine the effects of the increased fragility of hepatic mitochondria from ethanol-fed rats. Included are measurements os hepatic mitochondria turnover. The above will be combined with investigations of ethanol-related alterations in liver energy metabolism carried out with hepatocytes and in situ liver. The degree to which the energy state of the liver is decreased by chronic ethanol consumption will be investigated in intact liver in situ and in hepatocytes, utilizing 31P NMR spectroscopy to determine cellular phosphorylation potential. Ethanol-elicited functional alterations in the mitochondrion in vivo will be assessed by measurements of the protonmotive force generated by mitochondria inside the hepatocyte.