CULTURED HEPATOCYTES AS A MODEL OF ALCOHOL METABOLISM

在酒精性肝病动物模型的发展过程中， 很少有人把注意力集中在产生一种新的 体外模型采用肝细胞原代培养。因此，它是 这项研究的总体目标是检查控制和监管 对培养的大鼠肝细胞乙醇代谢的影响。使用这个 模型中，我们建议检验：(1)延长培养周期 与酒精接触有关。目前，肝细胞可以 只能在含酒精的培养基中培养7-9天， 鉴于我们已经成功地将它们保持在无酒精状态 中等期限，最长1个月。条件将被修改以延长 与含酒精培养基相联系的时期 几种方法：改变培养基02和C02 可用性，以及细胞附着的底物。(2)审查 它们对乙醇和乙醛代谢的调节 细胞培养至少7天以上。关于《红楼梦》的贡献研究 我们将评估乙醇代谢的三条已知途径。 同样的可能性是乙醇可以诱导出它自己 新陈代谢。乙醇代谢的代谢后果 脂肪肝的发生及对血清蛋白的影响 将对合成和分泌进行评估。规管 乙醛代谢及其长期接触对机体的影响 乙醇对乙醇和乙醛脱氢酶活性的影响 调查过了。(3)上述所有研究均会使用 正常大鼠肝细胞。这些研究将与 慢性酒精喂养大鼠细胞培养的研究 乙醇的存在或不存在，以检查 保持/消除耐受性。(4)诱导 和/或酒精P-450的维护。除了乙醇之外 本身，我们将使用吡唑、4-甲基吡唑或 异烟肼维持或诱导酒精P-450。单元格也会 从用这些制剂处理过的动物身上培养出来。(5) 氧自由基和脂质过氧化在心肌梗死中的可能作用 将评估与乙醇相关的肝脏毒性。 氧自由基的产生、脂质过氧化和对 长期接触乙醇后的氧化应激 下定决心。预计对这些问题进行检查 在严格控制的体外条件下的参数将导致 以更好地了解其发病机制的某些方面。 长期酗酒导致的肝病。

In the development of animal models of alcoholic liver disease, little attention has focused on the possibilities of generating an in vitro model using liver cells in primary culture. It is therefore the overall aim of this study to examine the control and regulation of ethanol metabolism in cultured rat hepatocytes. Using this model, we propose to test: (1) Extension of the culture period associated with ethanol exposure. At present, hepatocytes can only be cultured in alcohol containing medium for 7-9 days, whereas we have successfully maintained them in alcohol free medium for up to 1 month. Conditions will be modified to extend the period associated with alcohol containing culture medium in several ways: alterations to the culture medium, 02 and C02 availability, and substrate for cell attachment. (2) To examine the regulation of ethanol and acetaldehyde metabolism by these cells over at least 7 days culture. Studies on th contributions of the three known pathways of ethanol metabolism will be assessed as will be the possibility that ethanol can induce its own metabolism. Metabolic consequences of ethanol metabolism such as the generation of fatty liver and effect on serum protein synthesis and secretion will be evaluated. Regulation of acetaldehyde metabolism and the effect of prolonged exposure to ethanol on alcohol and aldehyde dehydrogenase activities will be investigated. (3) All of the above studies will be carried out using hepatocytes from normal rats. These studies will be paralleled by studies involving cells from chronic alcohol-fed rats cultured in the presence or absence of ethanol, to examine the maintenance/disappearance of tolerance. (4) The induction of and/or maintenance of the alcohol P-450. In addition to ethanol itself, we will use ligands such as pyrazole, 4-methylpyrazole or isoniazid to maintain or induce the alcohol P-450. Cells will also be cultured from animals pretreated with these agents. (5) The possible role of oxygen radicals and lipid peroxidation in the hepatotoxicity associated with ethanol will be evaluated. Generation of oxyradicals, lipid peroxidation and sensitivity to oxidative stress after prolonged exposure to ethanol will be determined. It is anticipated that examination of these parameters under carefully controlled conditions in vitro will lead to a better understanding of some aspects of the pathogenesis of liver disease following prolonged alcohol abuse.