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：
7879924
负责人：
ARTHUR I CEDERBAUM
金额：
$ 39.85万
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/7879924
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 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 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 public health relevance 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）：酒精会导致细胞损伤的机制尚不清楚。大量研究重点的主要机制是脂质过氧化和氧化应激在酒精毒性中的作用。已经提出许多途径在乙醇如何诱导“氧化应激”（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，尤其是TNF1和CYP2E1诱导的情况下发挥关键作用。这些途径并不彼此不包括，但是，它们之间的关联和相互作用，尤其是在体内条件下，仍有待评估和澄清。我们假设体内CYP2E1诱导中增加的氧化应激增加了肝细胞对TNF1诱导的肝毒性的敏感性。我们建议氧化剂，例如过氧亚硝酸盐，诸如JNK和/或p38 MAPK之类的MAP激酶，NF-KB的失活以及线粒体功能障碍是CYP2E1-TNF1的下游介体。我们还假设CYP2E1的诱导在体内用酒精治疗增强TNF1肝毒性的机制中起着核心作用。由于CYP2E1和TNF1被认为是酒精性肝损伤发展的关键危险因素，因此在体内的可能相互作用和增强其作用对于评估很重要。 AIM 1将通过诱导CYP2E1评估TNF1诱导的肝损伤的增强。 TNF1或生理盐水将用于具有基底水平CYP2E1的盐水处理的小鼠，或具有高水平CYP2E1的吡唑治疗的小鼠，而CYP2E1敲除小鼠，没有CYP2E1。将给出各种浓度的TNF1，在TNF1挑战后的不同时间杀死小鼠。我们将分析以下内容：肝毒性，氧化/亚硝化应激，NF-KB的激活，MAP激酶的激活，线粒体功能障碍。 AIM 2将评估CYP2E1在TNF1诱导乙醇诱导的肝损伤增强中的作用，并确定CYP2E1-乙醇TNF1相互作用的下游靶标。酒精摄入的暴饮暴食模型，然后是TNF1或盐水挑战，将是评估的急性乙醇模型。对于慢性乙醇喂养，将为2、4或6周提供对照或乙醇lieber-decarli饮食，然后对TNF1（或盐水）进行挑战。与上述类似的测定将进行。为了证明CYP2E1负责吡唑和乙醇模型的增强损伤，将使用CYP2E1基因敲除小鼠和人源化CYP2E1敲除小鼠。 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相互作用。后者可能具有潜在的治疗价值。 PHS 398/2590（修订版09/04，重新发行4/2006）页面延续格式页面