INTERACTION OF ETHANOL WITH HEPATIC MICROSOMES

在建立了微粒体乙醇氧化系统(Meos)为 不同于酒精脱氢酶(ADH)和过氧化氢酶的实体， 申请人现在希望回答所涉及的机制问题，并 MEOS在体内外乙醇代谢中的作用。具体来说， 乙醇诱导的细胞色素P-450将从大鼠体内纯化， 鹿鼠和狒狒。我们将对纯化的蛋白(S)进行鉴定 分子量、光谱、底物专一性和多肽 映射。尽管已知它们的细胞色素P-450存在差异 物种，我们希望找到一个共同的变化，可能会揭示一些 乙醇诱导的改变的性质及其对乙醇AS的特异性 一种底物。我们还将确定羟基自由基的程度 参与乙醇诱导形式的乙醇氧化 细胞色素P-450。此外，我们会评估 细胞色素P-450导致其他底物的代谢改变，如 类固醇激素和维甲酸(维生素A的一种形式对 酗酒者的营养状况，致癌物质(可能的 酗酒者癌症发病率增加的重要性)和 工业溶剂和常用药物的肝毒性增强， 包括促进脂质过氧化(与发病机制相关 酒精性肝损伤)。评估MEO的相对重要性和 来确定它对代谢耐受性的贡献有多大 在慢性乙醇消费后发展起来，我们将比较 ADH+和ADH-分离的肝细胞的(~3H)木糖醇和(H_3)乙醇 鹿鼠。在同一实验模型中，我们将研究同位素效应。 为了将MeOS与其他酒精氧化区分开来 系统，无论是在分离的肝细胞还是在体内。我们的广泛目标是 研究是为了帮助我们理解酗酒者与 生化术语中的非酒精；具体地说，我们计划确定 酒精对微粒体细胞色素P-450的影响 饮酒改变了酗酒者对内源性和 外源性化合物可能会影响酒精诱导的肝脏 病理、预防和治疗。

After having established the microsomal ethanol oxidizing system (MEOS) as an entity distinct from alcohol dehydrogenase (ADH) and catalase, the applicant now wishes to address the question of the mechanism involved and the role of MEOS in ethanol metabolism in vitro and in vivo. Specifically, the ethanol-induced form of cytochrome P-450 will be purified from rats, deermice and baboons. We will characterize the purified protein(s) in terms of molecular weight, spectra, substrate specificity and peptide mapping. Despite known differences in the cytochrome P-450 of these species, we hope to find a common change that may shed some light on the nature of the ethanol-induced alteration and its specificity for ethanol as a substrate. We will also determine to what extent hydroxyl radicals are involved in ethanol oxidation mediated by the ethanol-induced form of cytochrome P-450. Furthermore, we will assess whether the changes in cytochrome P-450 result in altered metabolism of other substrates such as steroid hormones and retinoic acid (a form of vitamin A of significance for the nutritional status of the alcoholic), carcinogens (of possible importance for the enhanced incidence of cancers in the alcoholic) and in enhanced hepatotoxicity of industrial solvents and commonly used drugs, including promotion of lipid peroxidation (of relevance to the pathogenesis of alcoholic liver injury). To assess the relative importance of MEOS and to determine how much it contributes to the metabolic tolerance that develops after chronic ethanol consumption, we will compare the fate of (3H) xylitol and (H3) ethanol in hepatocytes isolated from ADH+ and ADH- deermice. In the same experimental model we will study the isotope effect of deuterium to differentiate MEOS from the other alcohol oxidizing systems, both in isolated hepatocytes and in vivo. The broad aim of our study is to contribute to our understanding of how alcoholics differ from non-alcoholics in biochemical terms; specifically we plan to determine to what extent changes produced in the microsomal cytochrome P-450 by alcohol consumption alters the response of the alcoholic to endogenous and exogenous compounds in a way that may affect alcohol induced liver pathology, its prevention and its treatment.