Effect of ethanol on Kupffer cell/hepatocyte interactions and lipid metabolism

乙醇对库普弗细胞/肝细胞相互作用和脂质代谢的影响

• 批准号: 7857922
• 负责人: Suthat Liangpunsakul
• 金额: $ 13.18万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-15 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7857922
• 关键词: Alcoholic Fatty Liver; Alcoholic Liver Diseases; Binding Proteins; CCL2 gene; Cells; Cellular biology; Chronic; Clinical Medicine; Coculture Techniques; Conditioned Culture Media; Endotoxemia; Endotoxins; Energy Metabolism Pathway; Ethanol; Fatty Acids; Funding; Goals; Hepatocyte; Immunology; Inflammatory; Knowledge; Kupffer Cells; Laboratories; Lipopolysaccharides; Liver diseases; Messenger RNA; Molecular Biology; Pathogenesis; Patients; Phosphorylation; Physicians; Primary carcinoma of the liver cells; Public Health; Research; Response Elements; Role; Scientist; Signal Transduction; Sterols; TNF gene; Testing; Training; Very low density lipoprotein; Work; adenylate kinase; alcohol effect; career; cytokine; design; fatty acid oxidation; genetic regulatory protein; hepatoma cell; lipid metabolism; macrophage; mortality; problem drinker; professor; response; skills; thyroid hormone receptor associated protein 220; treatment effect; Alcoholic Fatty Liver; Alcoholic Liver Diseases; Binding Proteins; CCL2 gene; Cells; Cellular biology; Chronic; Clinical Medicine; Coculture Techniques; Conditioned Culture Media; Endotoxemia; Endotoxins; Energy Metabolism Pathway; Ethanol; Fatty Acids; Funding; Goals; Hepatocyte; Immunology; Inflammatory; Knowledge; Kupffer Cells; Laboratories; Lipopolysaccharides; Liver diseases; Messenger RNA; Molecular Biology; Pathogenesis; Patients; Phosphorylation; Physicians; Primary carcinoma of the liver cells; Public Health; Research; Response Elements; Role; Scientist; Signal Transduction; Sterols; TNF gene; Testing; Training; Very low density lipoprotein; Work; adenylate kinase; alcohol effect; career; cytokine; design; fatty acid oxidation; genetic regulatory protein; hepatoma cell; lipid metabolism; macrophage; mortality; problem drinker; professor; response; skills; thyroid hormone receptor associated protein 220; treatment effect

DESCRIPTION (provided by applicant): Alcoholic liver disease (ALD) continues to be a major public health problem and remains the leading cause of mortality in patients with chronic liver disease. Ethanol disrupts the pathways of energy metabolism by inhibiting AMP kinase (AMPK), and interferes with the activity of sterol response element binding protein (SREBP) and peroxisome proliferator activated receptor (PPAR), leading to alcoholic steatosis. Ethanol can induce endotoxemia. Lipopolysaccharide (LPS), a component of endotoxin, can stimulate Kupffer cells (KC), to produce cytokines. We hypothesize that LPS stimulates KC to release TNFa, which is central to the pathogenesis of alcoholic fatty liver disease through its effect on PPARa, SREBP, and AMPK. We will also study the role of other cytokines released by the macrophages (i.e., MCP-1) on these regulatory proteins. We plan to test our hypothesis by pursuing the following specific aims. 1) Determine the effect of exogenous TNFa or LPS in the presence or absence of ethanol on the function of PPARa, SREBP-1, and AMPK in McA-RH7777 hepatoma cells and isolated hepatocytes, 2) Determine the effect of LPS, in the presence or absence of ethanol, on the ability of macrophages, RAW 264.7 cells or KC, to modify the function of PPARa, SREBP-1, and AMPK in hepatoma cells and hepatocytes, 3) Determine the effects of co-culture of RAW 264.7 cells (or KC) with McA-RH7777 cells (or isolated hepatocytes) on lipid metabolism of the hepatoma cells (or isolated hepatocytes), and 4) Determine the ability of hepatoma cells or hepatocytes to modify the response of RAW cells or KC to release TNFa when stimulated by LPS, in the presence or absence of ethanol. The candidate is as Assistant Professor of Clinical Medicine. To date the candidate has trained in the laboratory of Dr. David Crabb, acquiring basic molecular biology knowledge and laboratory skills. This application is a logical extension of the work in this laboratory designed to allow the candidate to develop a basic understanding of the roles of cytokines in the pathogenesis of ALD. The candidate will develop new research skills by working with a number of experts in Kupffer cell biology, lipid metabolism, and TNF signaling, as well as through course work in molecular biology and immunology. By acquiring the knowledge and skills outlined in this proposal, the candidate hopes to fulfill his career goal of becoming a well-trained and funded physician scientist.

描述（由申请人提供）：酒精性肝病（ALD）仍然是一个主要的公共卫生问题，并且仍然是慢性肝病患者死亡的主要原因。乙醇通过抑制AMP激酶（AMPK）来破坏能量代谢的途径，并干扰固醇反应元件结合蛋白（SREBP）和过氧化物酶体增殖剂活化受体（PPAR）的活性，导致酒精性脂肪变性。乙醇可诱导内毒素血症。脂多糖（LPS）是内毒素的一种成分，可以刺激库普弗细胞（KC）产生细胞因子。我们假设LPS刺激KC释放TNFA，这是酒精脂肪肝疾病的发病机理，其对PPARA，SREBP和AMPK的影响。我们还将研究巨噬细胞（即MCP-1）在这些调节蛋白上释放的其他细胞因子的作用。我们计划通过追求以下特定目标来检验我们的假设。 1）确定在MCA-RH7777肝癌细胞中PPARA，SREBP-1和AMPK功能的外源TNFA或LPS对PPARA，SREBP-1和AMPK功能的影响，并分离出肝细胞，2）确定LPS的作用，确定LPS的作用，在MACRAPPAR的功能或不存在MACR的功能中，par offer的作用264.7。 SREBP-1和肝癌细胞和肝细胞中的AMPK，3）确定RAW 264.7细胞（或KC）与MCA-RH7777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777777元培养的影响（或孤立的肝细胞）对肝瘤细胞的脂质代谢（或分离的Hepatocytes and Hepat to）的影响。在存在或不存在乙醇的情况下，通过LPS刺激时，将原细胞或KC的响应修改对释放TNFA的响应。候选人是临床医学助理教授。迄今为止，候选人已经在David Crabb博士的实验室中培训，并获得了基本的分子生物学知识和实验室技能。该应用是该实验室中该作品的逻辑扩展，旨在使候选人能够对细胞因子在ALD发病机理中的作用产生基本了解。候选人将通过与库普弗细胞生物学，脂质代谢和TNF信号的许多专家以及通过分子生物学和免疫学的课程工作来发展新的研究技能。通过获得该提案中概述的知识和技能，候选人希望实现自己的职业目标，即成为一名训练有素且资助的医师科学家。