ETHANOL AND MAP KINASE CASCADE

乙醇和 MAP 激酶级联

• 批准号: 2679989
• 负责人: Shivendra D Shukla
• 金额: $ 16万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2679989
• 关键词: acute phase protein; alpha 1 acid glycoprotein; amyloid proteins; animal genetic material tag; cell nucleus; enzyme activity; enzyme induction /repression; ethanol; intracellular transport; isozymes; laboratory rat; liver cells; liver pharmacology; mitogen activated protein kinase; phospholipase A2; phosphoproteins; protein metabolism; protein tyrosine kinase; tissue /cell culture

DESCRIPTION: (Adapted from the Investigator's Abstract) The long-term OBJECTIVE of the project is to determine the mechanism and consequences of ethanol effects on cell signaling mediated via tyrosine kinase (TK) and mitogen activated protein kinases (MAPK). Preliminary results suggest that ethanol modulated both kinases. In liver cells (BNLCL2) and rat hepatocytes, ethanol dramatically potentiates MAPK activation by serum. This is not due to upregulation of MAPK protein or physical interaction between ethanol and MAPK (p42 and p44) and therefore may be due to modulation on post-receptor signaling steps. The central hypothesis is that ethanol modulates serum-stimulated TK and Ras-Raf-MAPKK cascade leading to potentiation of MAPK and its translocation into the nucleus. Rat hepatocytes will be the major model of the proposal. There are three aims: Aim 1. To determine pharmacology of serum stimulated tyrosine and MAP kinases in rat hepatocytes. Serum activated MAPK will be determined in ethanol (10-100 mM) treated (2-4 days) rat hepatocytes. Next these parameters will be tested in hepatocytes from control (CH) and ethanol-fed (EH) rats. Aim II. To determine the mechanism of ethanol potentiation of serum stimulated MAPK. Involvement of Ras activation (active Ras GTP form) and Ras-Raf-interaction in this ethanol effect will be investigated. Aim III. To determine nuclear translocation of MAPK by ethanol and its downstream relevance. MAPK levels in the nucleus of ethanol-treated hepatocytes (Aim1) will be determined. Next its level in the nuclei of CH and EH (+ serum stimulation) will be established. Consequences of the MAPK potentiation on c-PLA2 and of its nuclear translocation on the expression of acute phase proteins will be ascertained. Significance: This project will establish the mechanism and molecular target(s) involved in ethanol potentiation of serum stimulated MAPK. New knowledge on the nuclear effects of ethanol on MAPK will be gained. Ethanol potentiated MAP Kinase may emerge an as important link between cytosol and nucleus to elicit nuclear responses to ethanol. Such molecular elucidation of the ethanol actions offers the potential to design therapeutic tools to detect, treat, and prevent ethanol-induced liver injury.

描述：（根据调查员的摘要改编）长期 该项目的目的是确定 乙醇对通过酪氨酸激酶（TK）和 有丝分裂原活化蛋白激酶（MAPK）。 初步结果表明 乙醇调节了两种激酶。 在肝细胞（BNLCL2）和大鼠中 肝细胞，乙醇急剧通过血清激活MAPK。 这不是由于MAPK蛋白或物理相互作用的上调 在乙醇和MAPK（p42和p44）之间，因此可能是由于 对受体后信号传导步骤进行调制。 中心假设是 乙醇调节血清刺激的TK和RAS-RAF-MAPKK级联反应 MAPK的增强及其转移到核中。 鼠 肝细胞将是该提案的主要模型。 有三个目标： 目的1。确定血清刺激酪氨酸的药理学和地图 大鼠肝细胞中的激酶。 血清激活的MAPK将在 乙醇（10-100毫米）治疗（2-4天）大鼠肝细胞。 接下来这些 参数将在对照（CH）和乙醇喂养的肝细胞中进行测试 （EH）大鼠。 目标II。 确定乙醇增强的机理 血清刺激的MAPK。 RAS激活的参与（活动RAS GTP形式） 将研究这种乙醇效应中的RAS-RAF相互作用。 目的 iii。 确定乙醇及其MAPK的核易位 下游相关性。 乙醇处理细胞核中的MAPK水平 将确定肝细胞（AIM1）。 接下来它的CH核中的水平 将建立EH（+血清刺激）。 MAPK的后果 C-Pla2及其核转运的增强 将确定急性相蛋白。 意义：这个项目将 建立乙醇涉及的机制和分子靶标 血清刺激的MAPK的增强。 有关核影响的新知识 将获得MAPK上乙醇的乙醇。 乙醇增强的地图激酶可能 出现胞质和核之间的重要联系以引起核 对乙醇的反应。 这种分子阐明乙醇作用 提供设计治疗工具来检测，治疗和 防止乙醇诱导的肝损伤。