Role of Alcohol Metabolism in Alcoholic Chronic Pancreatitis

酒精代谢在酒精性慢性胰腺炎中的作用

• 批准号: 9897449
• 负责人: BHUPENDRA S KAPHALIA
• 金额: $ 34.88万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-20 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9897449
• 关键词: 5' -AMP-activated protein kinase; Acinar Cell; Agonist; Alcohol abuse; Alcohol consumption; Alcohol dehydrogenase; Alcoholic Pancreatitis; Animal Model; Animals; Attenuated; Cell Culture Techniques; Cessation of life; Chronic; Clinical Research; Cyclic AMP-Dependent Protein Kinases; Deer Mouse; Diabetes Mellitus; Diet; Disease; Dose; Early Diagnosis; Esters; Ethanol; Ethanol Metabolism; Etiology; Exocrine pancreas; Fatty Acids; Fibrosis; Hepatic; Homeostasis; Human; Impairment; In Vitro; Incubated; Individual; Inflammation; Inflammatory; Injury; Lead; Lipids; Liquid substance; Liver; Malignant neoplasm of pancreas; Metabolic; Metabolism; Modeling; Molecular Target; Nature; Oral; Oxides; Pain; Pancreas; Pancreatic Injury; Pancreatitis; Pathogenesis; Patients; Pharmacology; Phenotype; Play; Prevention; Protein Isoforms; Research; Research Personnel; Ribonucleotides; Role; Signal Transduction; Testing; Therapeutic Intervention; Toxic effect; Variant; alcohol exposure; alcohol induced pancreatic injury; alcoholic chronic pancreatitis; attenuation; cell injury; chronic alcohol ingestion; chronic pancreatitis; clinically significant; comorbidity; cost; disorder prevention; endoplasmic reticulum stress; experience; fatty acid oxidation; feeding; fomepizole; in vivo; inhibitor/antagonist; innovation; lipophilicity; mouse model; novel; oxidation; problem drinker; therapeutic target

Abstract Chronic alcohol abuse is a single most major etiology of chronic pancreatitis, a serious inflammatory disorder of exocrine pancreas leading to loss of pancreatic functions and multiple co-morbidities including diabetes and pancreatic cancer. Therefore, a better understanding of mechanism and metabolic basis of alcoholic chronic pancreatitis (ACP) is of great clinical significance for identifying its therapeutic targets for early detection/prevention of the disease. Majority (~90%) of ingested alcohol (ethanol) is metabolized in the liver via alcohol dehydrogenase (ADH). However, the inhibition of hepatic ADH during chronic alcohol abuse facilitates formation of fatty acid ethyl esters (FAEEs, nonoxidative metabolites of ethanol) by several folds in the pancreas frequently damaged during chronic alcohol abuse. These esters are known to cause injury to pancreatic acinar cells in vitro as well as in vivo. Using hepatic ADH-deficient (ADH-) deer mice fed 3.5% ethanol via Lieber-DeCarli liquid diet, we found formation of fatty pancreas and several fold increases for pancreatic FAEEs, and endoplasmic reticulum (ER) stress and injury. Additionally, we also found inactivation of AMP-activated protein kinase (AMPK)α, which regulates lipid homeostasis via controlling lipid synthesis and β-oxidation of fatty acids, in freshly isolated human pancreatic acinar cells exposed to ethanol in vitro, and in the pancreas of ADH- vs. hepatic normal ADH (ADH+) deer mice after chronic ethanol feeding. Together, these preliminary findings led our central hypothesis that chronic ingestion of ethanol and its nonoxidative metabolism under hepatic ADH inhibition deactivates pancreatic AMPKα resulting into formation of a large quantities of FAEEs in the pancreas, contributing to pathogenesis of ACP. This hypothesis will be tested by establishing progressive pancreatic injury in ADH- deer mice fed ethanol for 1 and 3 months (aim 1), and that chronic ethanol feeding inactivates AMPKα, promotes increased formation of FAEEs resulting into progressive pancreatic injury in ADH- deer mice (aim 2). The in vivo findings will be validated in primary human pancreatic acinar cells. Finally, role of FAEEs in ethanol-induced pancreatic acinar cell injury will be established by using ADH- deer mice and primary human pancreatic acinar cells (aim 3). We expect a progressive ethanol-induced pancreatic injury in our deer mouse model, and to identify the role of endogenous FAEEs in ethanol-induced pancreatic acinar cell injury. Our project is innovative because we are using a hepatic ADH- deer mouse model (a natural variant of hepatic ADH deficiency) and freshly isolated human pancreatic acinar cells to establish the metabolic basis and mechanism of ACP. This project will be benefited by a strong interdisciplinary team of investigators and their research experience with deer mouse and human pancreatic acinar cell culture models. Overall, our project should establish metabolic basis of ACP and identify molecular targets for an early detection/therapeutic intervention of ACP.

摘要 慢性酒精滥用是慢性胰腺炎的一个最主要的原因，慢性胰腺炎是一种严重的炎症性疾病 外分泌胰腺导致胰腺功能丧失和多种并发症，包括糖尿病和 胰腺癌。因此，对慢性酒精中毒的发病机制和代谢基础有了更好的认识 胰腺炎(ACP)对早期明确治疗靶点具有重要的临床意义。 疾病的检测/预防。大部分(~90%)摄入的酒精(乙醇)在肝脏中通过 乙醇脱氢酶(ADH)。然而，慢性酒精滥用期间对肝脏ADH的抑制有助于 脂肪酸乙酯(FAEEs，乙醇的非氧化代谢物)在 在长期酗酒期间，胰腺经常受损。众所周知，这些酯会对人体造成伤害 胰腺腺泡细胞的体外培养和体内培养。用肝脏ADH缺陷(ADH-)鹿小鼠饲喂3.5% 通过Lieber-DeCarli液体饮食，我们发现脂肪胰腺的形成，并增加了几倍 胰腺FAEs、内质网(ER)应激和损伤。此外，我们还发现了失活 AMPK的α，它通过控制脂质的合成和调节脂质的动态平衡 在乙醇暴露的体外新鲜分离的人胰腺腺泡细胞中，脂肪酸的β氧化，以及 ADH-和肝正常ADH(ADH+)鹿小鼠慢性酒精喂养后胰腺的变化。加在一起，这些 初步的发现引导了我们的中心假设，即慢性摄入乙醇及其非氧化性 肝细胞腺苷脱氢酶抑制作用下的代谢失活胰腺AMPKα导致 胰腺中有大量FAEs，参与了ACP的发病。这一假设将是 以酒精喂养1个月和3个月的ADH-鹿小鼠建立进行性胰腺损伤模型进行测试(目标1)， 慢性乙醇摄入使AMPKα失活，促进FAEs的形成，从而导致 ADH-鹿小鼠进行性胰腺损伤(目标2)。体内的发现将在原代人类身上得到验证 胰腺腺泡细胞。最后，FAEs在乙醇诱导的胰腺腺泡细胞损伤中的作用将是 用ADH-鹿小鼠和原代人胰腺腺泡细胞建立(目标3)。我们预计会有一个 乙醇致小鼠进行性胰腺损伤模型的建立及内源性的作用 FAEs在乙醇诱导的胰腺腺泡细胞损伤中的作用。我们的项目具有创新性，因为我们使用了 肝脏ADH-鹿小鼠模型(肝脏ADH缺乏症的自然变体)和新鲜分离的人 建立胰腺腺泡细胞ACP的代谢基础和作用机制。这个项目将会受益 由一支强大的跨学科调查团队及其对鹿、鼠和人的研究经验 胰腺腺泡细胞培养模型。总体而言，我们的项目应该建立ACP的代谢基础并确定 ACP早期发现/治疗干预的分子靶点。