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Role of CYP2E1 in TNFa-Ethanol -induced Liver Injury

CYP2E1 在 TNFa-乙醇诱导的肝损伤中的作用

基本信息

批准号：
8099762
负责人：
ARTHUR I CEDERBAUM
金额：
$ 38.31万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8099762
关键词：
3-nitrotyrosine Acetylcysteine Acute Alcohol consumption Alcoholic Liver Diseases Alcohols Antioxidants Apoptotic Binding Biological Assay CYP2E1 gene Cell Nucleus Chemosensitization Chronic Coupled Cyclosporine Development Diet Dissociation Ethanol Ethanol toxicity Family Gene Targeting Generations Goals Health Hepatocyte Hepatology Hepatotoxicity Human Injury Knockout Mice Lipid Peroxidation Liver MAP Kinase Gene MAP Kinase Kinase Kinase MAPK phosphatase MAPK14 gene MAPK8 gene MAPK9 gene Mediator of activation protein Membrane Potentials Mitochondria Mitogen-Activated Protein Kinase Kinases Mitogen-Activated Protein Kinases Modeling Molecular Mus NF-kappa B Necrosis Nuclear Oxidants Oxidative Stress Oxygen Consumption Pathway interactions Permeability Peroxonitrite Play Production Proteins Pyrazoles Reactive Oxygen Species Research Risk Factors Role Saline Signal Pathway Signal Transduction Site Staging Superoxides Swelling TNF gene Testing Therapeutic Thioredoxin Time Toxic effect adduct alcohol abuse therapy cell injury cytokine feeding in vivo inhibitor/antagonist killings member mitochondrial dysfunction nitrosative stress oxidative damage prevent research study response restoration synergism

项目摘要

DESCRIPTION (provided by applicant): The mechanism(s) by which alcohol causes cell injury are still not clear. A major mechanism that is a focus of considerable research is the role of lipid peroxidation and oxidative stress in alcohol toxicity. Many pathways have been suggested to play a key role on how ethanol induces "oxidative stress", including mitochondrial dysfunction, activation of MAP kinases, ethanol-induced increases in cytokine formation, especially TNF1 and induction of CYP2E1. These pathways are not exclusive of each other, however, associations and interactions between them, especially under in vivo conditions, remain to be evaluated and clarified. We hypothesize that increased oxidative stress from CYP2E1 induction in vivo sensitizes hepatocytes to TNF1-induced hepatotoxicity. We propose that oxidants such as peroxynitrite, activation of MAP kinases such as JNK and/or P38 MAPK, inactivation of NF-kB and mitochondrial dysfunction are downstream mediators of the CYP2E1- TNF1 potentiated hepatotoxicity. We also hypothesize that induction of CYP2E1 plays a central role in mechanisms by which in vivo treatment with alcohol potentiates TNF1 hepatotoxicity. Since CYP2E1 and TNF1 are considered key risk factors in the development of alcoholic liver injury, possible interactions and potentiation of their actions in vivo is important to evaluate. AIM 1 will evaluate the potentiation of TNF1-induced liver injury by induction of CYP2E1. TNF1 or saline will be administrated to either saline-treated mice with basal levels of CYP2E1, or pyrazole-treated mice with high levels of CYP2E1, and to CYP2E1-knockout mice with no CYP2E1. Various concentrations of TNF1 will be given and mice killed at various times after TNF1 challenge. We will assay for the following: Hepatotoxicity, oxidative/nitrosative stress, activation of NF-kB, activation of MAP kinases, mitochondrial dysfunction. AIM 2 will evaluate the role of CYP2E1 in the potentiation of TNF1-induced liver injury by ethanol and to identify downstream targets for the CYP2E1-ethanol- TNF1 interactions. A binge model of alcohol intake followed by TNF1 or saline challenge will be the acute ethanol model evaluated. For chronic ethanol feeding, mice will be fed a control or an ethanol Lieber-DeCarli diet for 2, 4 or 6 weeks, followed by challenge with TNF1 (or saline). Assays similar to the above will be carried out. To prove that CYP2E1 is responsible for the potentiated injury in the pyrazole and ethanol models, CYP2E1 knockout mice and humanized CYP2E1 knockin mice will be used. PUBLIC HEALTH RELEVANCE: We believe that these experiments will help to clarify how two independent risk factors believed to be important for alcohol-induced liver injury, CYP2E1 and TNF1 interact to promote this liver injury, will provide clear evidence for a role for CYP2E1 in acute and chronic alcohol/ TNF1 potentiated liver injury, and help to identify downstream mediators, signaling pathways and targets of the CYP2E1-ethanol- TNF1 interactions. The latter may have potential therapeutic value. PHS 398/2590 (Rev. 09/04, Reissued 4/2006) Page Continuation Format Page

描述（由申请人提供）：酒精导致细胞损伤的机制仍不清楚。作为大量研究的焦点的一个主要机制是脂质过氧化和氧化应激在酒精毒性中的作用。许多途径被认为在乙醇如何诱导“氧化应激”中起关键作用，包括线粒体功能障碍、MAP激酶的激活、乙醇诱导的细胞因子形成的增加，特别是TNF 1和CYP 2 E1的诱导。这些途径并不相互排斥，然而，它们之间的关联和相互作用，特别是在体内条件下，仍有待评估和澄清。我们假设体内CYP 2 E1诱导引起的氧化应激增加使肝细胞对TNF 1诱导的肝毒性敏感。我们认为氧化剂如过氧亚硝酸盐、MAP激酶如JNK和/或P38 MAPK的激活、NF-kB的失活和线粒体功能障碍是CYP 2 E1-TNF 1增强肝毒性的下游介质。我们还推测，CYP 2 E1的诱导在体内酒精治疗增强TNF 1肝毒性的机制中起着重要作用。由于CYP 2 E1和TNF 1被认为是酒精性肝损伤发展的关键风险因素，因此评价其体内作用的可能相互作用和增强作用非常重要。目的1将评估通过诱导CYP 2 E1增强TNF 1诱导的肝损伤。将TNF 1或盐水给予具有基础水平的CYP 2 E1的盐水处理的小鼠，或具有高水平的CYP 2 E1的吡唑处理的小鼠，以及无CYP 2 E1的CYP 2 E1敲除小鼠。将给予不同浓度的TNF 1，并在TNF 1激发后的不同时间处死小鼠。我们将分析以下内容：肝毒性、氧化/亚硝化应激、NF-κ B活化、MAP激酶活化、线粒体功能障碍。目的2将评估CYP 2 E1在乙醇增强TNF-1诱导的肝损伤中的作用，并确定CYP 2 E1-乙醇-TNF-1相互作用的下游靶点。酒精摄入后TNF 1或生理盐水激发的狂欢模型将是评价的急性乙醇模型。对于慢性乙醇喂养，将小鼠喂养对照或乙醇Lieber-DeCarli饮食2、4或6周，然后用TNF 1（或盐水）攻击。将进行与上述类似的试验。为了证明CYP 2 E1是吡唑和乙醇模型中增强损伤的原因，将使用CYP 2 E1敲除小鼠和人源化CYP 2 E1敲入小鼠。公共卫生相关性：我们相信，这些实验将有助于澄清两个独立的危险因素，认为是重要的酒精诱导的肝损伤，CYP 2 E1和TNF 1相互作用，以促进这种肝损伤，将提供明确的证据，CYP 2 E1在急性和慢性酒精/TNF 1增强肝损伤的作用，并有助于确定下游介质，信号通路和CYP 2 E1-乙醇-TNF 1相互作用的目标。后者可能具有潜在的治疗价值。PHS 398/2590（2004年9月修订，2006年4月重新印发）