ETHANOL INDUCTION OF UDP-GLUCURONYLTRANSFERASE

UDP-葡萄糖醛酸转移酶的乙醇诱导

• 批准号: 2043492
• 负责人: Garold S Yost
• 金额: $ 12.39万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-02-01 至 1995-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2043492
• 关键词: alcoholic beverage consumption; alcoholism /alcohol abuse; alcoholism /alcohol abuse chemotherapy; complementary DNA; drug interactions; drug metabolism; enzyme induction /repression; enzyme mechanism; enzyme structure; enzyme substrate; ethanol; glucuronides; glucuronosyltransferase; high performance liquid chromatography; human tissue; immunoprecipitation; isozymes; laboratory rabbit; messenger RNA; nucleic acid probes; protein sequence; sheep; thin layer chromatography; western blottings

APPLICANT'S ABSTRACT: Chronic alcohol consumption changes the therapeutic efficacies and pharmacokinetics of a number of drugs. Usually, these changes are manifest as increased metabolism of the drugs leading to shorter half-lives, increased clearance, and decreased plasma concentrations. These changes can lead to inadequate therapeutic doses. Such effects may primarily be observed in the clinical treatment of alcoholics, but may be important in the effects of moderate alcohol consumption on drug efficacies. Glucuronidation of many of these drugs is the major or only route of elimination, and induction of UDP-glucuronosyltransferase isozymes could lead to significant increases in the clearance of the drugs. Chronic alcohol exposure induces the production of a unique UDP-glucuronosyltransferase (GT) isozyme in rabbit hepatic microsomes, and this biochemical effect is manifest in the increased clearance, via glucuronidation, of morphine and oxazepam in the animals. The major goal of this research is to determine the precise molecular events and dose requirements that control this induction process in rabbits, and to demonstrate that these processes also occur in man. Realization of this goal will be accomplished through the following methods: 1) the production and use of antibodies to the rabbit ethanol-induced GT in immunoblots and immunoprecipitation studies with ethanol-induced rabbit microsomes and purified isozymes to determine the time course and dose requirements for induction; 2) preparation and sequencing of rabbit hepatic cDNA clones of the DNA coding for the ethanol-induced GT and use of cDNA probes to screen rabbit and human mRNA levels to determine the molecular events that are responsible for production of the isozyme; and 3) purification and characterization of the ethanol-induced GT protein(s) and cDNAs from human liver tissue. The long-term objectives of this research are to establish the existence of ethanol-induced UDP-glucuronosyltransferase isozymes in experimental animals and humans, characterize the isozymes and provide information about the molecular events that are responsible for their production, and relate these biochemical events to the treatment of humans, after chronic exposure to ethanol, with drugs that are metabolized by these GT isozymes. These studies represent a multifaceted approach on the biochemical, molecular biological, and clinical levels that will provide significant new information about the effects of chronic alcohol consumption on an important human drug metabolism enzyme.

申请人摘要：长期饮酒改变了治疗 许多药物的药效和药代动力学。 通常这些 变化表现为药物代谢增加， 半衰期缩短，清除率增加，血浆 浓度的 这些变化可能导致治疗剂量不足。 这种作用主要可以在临床治疗中观察到， 酗酒者，但可能是重要的影响，适度的酒精 药物功效的消耗。 许多药物的葡萄糖醛酸化是 主要或唯一的消除途径，以及诱导 UDP-葡萄糖醛酸基转移酶同工酶可导致 清除毒品 慢性酒精暴露诱导 一种独特的兔UDP-葡萄糖醛酸基转移酶（GT）同工酶的产生 肝微粒体，这种生物化学作用表现在 通过葡萄糖醛酸化增加吗啡和奥沙西泮在 动物 这项研究的主要目的是确定精确的 控制这一诱导过程的分子事件和剂量要求 在兔子身上，并证明这些过程也发生在人类身上。 实现这一目标将通过以下途径实现 方法：1）兔抗人源性抗体的制备及应用 乙醇诱导的GT的免疫印迹和免疫沉淀研究， 乙醇诱导的兔微粒体和纯化的同工酶来确定 诱导的时间过程和剂量要求; 2）准备和 兔肝cDNA克隆的DNA编码的测序 乙醇诱导的GT和使用cDNA探针筛选兔和人mRNA 水平来确定导致 同工酶的产生;和3）同工酶的纯化和表征。 乙醇诱导的GT蛋白和来自人肝组织的cDNA。 的 这项研究的长期目标是建立 乙醇诱导的尿苷二磷酸葡萄糖醛酸基转移酶同工酶 动物和人类，表征同工酶，并提供有关 负责其生产的分子事件， 这些生化事件对人类的治疗， 乙醇，与这些GT同工酶代谢的药物。 这些 研究代表了生物化学、分子生物学和生物学的多方面方法。 生物和临床水平，将提供重要的新的 关于长期饮酒对健康影响的信息 重要人体药物代谢酶。

项目成果

Purification and characterization of an ethanol-induced UDP-glucuronosyltransferase.

乙醇诱导的 UDP-葡萄糖醛酸基转移酶的纯化和表征。

• DOI: 10.1016/0003-9861(89)90157-4
• 发表时间: 1989
• 期刊: Archives of biochemistry and biophysics
• 影响因子: 3.9
• 作者: Hutabarat,RM;Yost,GS
• 通讯作者: Yost,GS