The role of Kupffer cells in alcohol-induced liver disease

库普弗细胞在酒精性肝病中的作用

基本信息

批准号：
9761401
负责人：
YASUKO IWAKIRI
金额：
$ 37.62万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-15 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9761401
关键词：
Adoptive Transfer Alcohol abuse Alcoholic Fatty Liver Alcoholic Hepatitis Alcoholic Liver Diseases Animals Anti-inflammatory B-Lymphocytes Cells Cirrhosis Clinical Data Development Diet Early Intervention Encapsulated Endoplasmic Reticulum Endothelial Cells Enhancers Ethanol Exhibits Fatty Liver Fibrosis Goals Hepatic Stellate Cell Hepatocyte Human In Vitro Inflammatory Inflammatory Response Injury Interleukin-1 beta Knock-out Knockout Mice Kupffer Cells Light Link Liver Liver diseases Molecular Mus NOS2A gene Nuclear Patients Play Primary carcinoma of the liver cells Proteins Reporting Research Role Severities Small Interfering RNA Specimen Structure TNF gene Testing Therapeutic alcohol response arginase base chronic alcohol ingestion effective therapy endoplasmic reticulum stress experimental study factor A in vivo liver injury macrophage nanoparticle non-alcoholic novel paracrine polarized cell prevent problem drinker response therapeutic target

项目摘要

SUMMARY Alcohol-induced liver disease is a significant clinical problem. Kupffer cells (liver resident macrophages) play crucial roles in the inflammatory responses of alcoholic liver disease. Macrophages have distinct functional states with pro-inflammatory M1 type and anti-inflammatory M2 type. The mechanisms that govern this classical polarization remain to be elucidated. The goals of this study are to: 1) Identify a novel molecular switch that determines M1 vs. M2 polarization in the context of ethanol- induced hepatic steatosis and injury and 2) Evaluate the potential of Kupffer cells as a therapeutic target. An endoplasmic reticulum (ER) resident protein, Nogo-B, also known as reticulon 4B, has been implicated in maintaining ER structure. In the liver, Nogo-B is restricted to non-parenchymal cells including Kupffer cells, liver sinusoidal endothelial cells and hepatic stellate cells, but not in hepatocytes. Our preliminary data demonstrate that Nogo-B levels correlate with the severity of alcoholic liver disease in patients. Nogo-B levels in Kupffer cells were positively associated with M1 polarization and negatively with M2 polarization in human liver specimens. In mice, the absence of Nogo-B resulted in significantly lower levels of hepatic steatosis and injury than wildtype (WT) mice in response to an ethanol diet. Kupffer cells from Nogo-B knockout (KO) mice showed significantly decreased expression of M1 markers, including inducible nitric oxide synthase (iNOS), interleukin 1β (IL1β) and tumor necrosis factor α (TNFα), but exhibited significantly increased M2 markers, such as CD163 and arginase-1, compared to their WT counterparts. Importantly, iNOS, IL1β and TNFα have been reported to enhance hepatic steatosis in alcoholic or non-alcoholic settings and are induced by nuclear factor kappa-light-chain-enhancer of activated B cells (NFkB). Nogo-B KO Kupffer cells exhibited significantly increased ER stress, a factor that induces M2 polarization. Based on these observations from human specimens and animal studies, we hypothesize that Nogo-B regulates Kupffer cell polarization and facilitates hepatic steatosis/injury in response to chronic ethanol consumption and that selective deletion of Nogo-B in Kupffer cells will reduce ethanol-induced hepatic injury. To test these hypotheses, we propose the following three aims: 1) Determine the mechanism by which Nogo-B facilitates M1 polarization of Kupffer cells in response to chronic ethanol consumption, 2) Determine the mechanism by which lack of Nogo-B facilitates M2 polarization of Kupffer cells in response to chronic ethanol consumption, and 3) Determine whether deletion of Nogo-B in Kupffer cells reduces hepatic steatosis and injury in ethanol-fed mice.

概括酒精引起的肝病是一个重要的临床问题。巨噬细胞）在酒精性肝病的炎症反应中发挥着至关重要的作用。具有不同的功能状态，分为促炎M1型和抗炎M2型。控制这种经典极化的机制仍有待阐明。这项研究的目标是。 1) 确定一种新颖的分子开关，可以在乙醇的背景下确定 M1 与 M2 极化诱导的肝脂肪变性和损伤，2) 评估库普弗细胞作为治疗药物的潜力目标。内质网 (ER) 驻留蛋白 Nogo-B，也称为网状蛋白 4B，已被参与维持 ER 结构在肝脏中，Nogo-B 仅限于非实质细胞。包括库普弗细胞、肝窦内皮细胞和肝星状细胞，但不包括我们的初步数据表明 Nogo-B 水平与肝细胞的严重程度相关。酒精性肝病患者 Kupffer 细胞中的 Nogo-B 水平与 M1 呈正相关。在人类肝脏标本中，M2 极化和负极化在小鼠中没有。 Nogo-B 导致肝脏脂肪变性和损伤水平显着低于野生型 (WT) 小鼠 Nogo-B 敲除 (KO) 小鼠的 Kupffer 细胞对乙醇饮食的反应表现出显着。 M1 标记物的表达，包括诱导型一氧化氮合酶 (iNOS)、白细胞介素 1β (IL1β) 和肿瘤坏死因子 α (TNFα)，但显示 M2 标记物显着增加，例如 CD163 和精氨酸酶-1 与它们的 WT 盟友相比，重要的是 iNOS、IL1β 和 TNFα 具有。据报道，在酒精或非酒精环境中会增强肝脂肪变性，并由以下因素引起活化 B 细胞 (NFkB) 的核因子 kappa 轻链增强子 (Nogo-B KO Kupffer 细胞)。显示显着增加的 ER 应力，这是诱导 M2 极化的一个因素。根据人类标本和动物研究的观察，我们勇敢地说 Nogo-B 调节库普弗细胞极化并促进慢性乙醇引起的肝脂肪变性/损伤消耗并且选择性删除库普弗细胞中的Nogo-B将减少乙醇诱导的肝损伤为了检验这些假设，我们提出以下三个目标：1）确定机制 Nogo-B 促进库普弗细胞的 M1 极化，以响应长期乙醇消耗，2) 确定缺乏 Nogo-B 促进 Kupffer 细胞 M2 极化的机制对慢性乙醇消耗的反应，以及 3) 确定 Kupffer 细胞中 Nogo-B 是否被删除减少乙醇喂养小鼠的肝脏脂肪变性和损伤。