Fructose Induced Regulation of Profibrogenic Factors in Hepatic Nonparenchymal Cells

果糖诱导的肝非实质细胞促纤维化因子的调节

• 批准号: 10373759
• 负责人: Arion Kennedy
• 金额: $ 18.45万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-24 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373759
• 关键词: Acetates; Acetyl Coenzyme A; Acetylation; Adult; Aldolase B; Automobile Driving; Biological Response Modifiers; Carbohydrates; Cell physiology; Cells; Child; Citric Acid Cycle; Consumption; Data; Development; Diabetes Mellitus; Diagnosis; Diet; Dietary intake; Discontinuous Capillary; Disease; Enzymes; Epidemic; Event; Extracellular Matrix Proteins; Fatty Liver; Fibrosis; Financial Support; Food Interactions; Foundations; Fructose; Funding Mechanisms; Gene Expression; Generations; Glucose; Glyceraldehyde; Glycogen; Goals; Hepatic; Hepatic Stellate Cell; Hepatocyte; Histones; IL1B gene; IL6 gene; Immune signaling; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Ketohexokinase; Kupffer Cells; Lipids; Liver; Liver Cirrhosis; Liver Fibrosis; Liver diseases; Lobular; Metabolic Pathway; Metabolism; Metalloproteases; Methanol; Monosaccharides; Nuclear Magnetic Resonance; Nutritional; Obesity; Outcome; Oxidative Stress; Pathogenesis; Pathology; Patients; Phenotype; Preparation; Prevention; Primary carcinoma of the liver cells; Production; Pyruvate; Regulation; Research; Research Design; Role; Saturated Fatty Acids; Small Interfering RNA; Societies; Source; Supplementation; System; TIMP1 gene; Therapeutic; Time; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinases; Transfection; Water; adult obesity; base; carbohydrate metabolism; dietary excess; drinking water; in vivo; inhibitor/antagonist; insight; lipid biosynthesis; liquid chromatography mass spectrometry; macrophage; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; obesity in children; protein expression; stellate cell; therapeutic development; therapeutic target; Acetates; Acetyl Coenzyme A; Acetylation; Adult; Aldolase B; Automobile Driving; Biological Response Modifiers; Carbohydrates; Cell physiology; Cells; Child; Citric Acid Cycle; Consumption; Data; Development; Diabetes Mellitus; Diagnosis; Diet; Dietary intake; Discontinuous Capillary; Disease; Enzymes; Epidemic; Event; Extracellular Matrix Proteins; Fatty Liver; Fibrosis; Financial Support; Food Interactions; Foundations; Fructose; Funding Mechanisms; Gene Expression; Generations; Glucose; Glyceraldehyde; Glycogen; Goals; Hepatic; Hepatic Stellate Cell; Hepatocyte; Histones; IL1B gene; IL6 gene; Immune signaling; In Vitro; Inflammation; Inflammation Mediators; Inflammatory; Ketohexokinase; Kupffer Cells; Lipids; Liver; Liver Cirrhosis; Liver Fibrosis; Liver diseases; Lobular; Metabolic Pathway; Metabolism; Metalloproteases; Methanol; Monosaccharides; Nuclear Magnetic Resonance; Nutritional; Obesity; Outcome; Oxidative Stress; Pathogenesis; Pathology; Patients; Phenotype; Preparation; Prevention; Primary carcinoma of the liver cells; Production; Pyruvate; Regulation; Research; Research Design; Role; Saturated Fatty Acids; Small Interfering RNA; Societies; Source; Supplementation; System; TIMP1 gene; Therapeutic; Time; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinases; Transfection; Water; adult obesity; base; carbohydrate metabolism; dietary excess; drinking water; in vivo; inhibitor/antagonist; insight; lipid biosynthesis; liquid chromatography mass spectrometry; macrophage; non-alcoholic fatty liver disease; nonalcoholic steatohepatitis; novel; obesity in children; protein expression; stellate cell; therapeutic development; therapeutic target

PROJECT SUMMARY NAFLD has become progressively more common in today's society impacting adults and children. High fructose corn syrup consumption is strongly correlated with diabetes, obesity and nonalcoholic fatty liver disease (NAFLD). Fructose consumption is two to three times higher in patients with NAFLD compared to controls and increases the presence of lobular inflammation and fibrosis. The major contributors of lobular inflammation and fibrosis are activated hepatic nonparenchymal cells (NPCs), which consist of hepatic stellate cells (HSCs), and Kupffer cells. Fructose metabolism has been investigated primarily in hepatocytes and is known to induce lipogenesis and oxidative stress. These events in hepatocytes have been shown to impact Kupffer cell function pushing the cells to an inflammatory phenotype. However, the distinct role of fructose metabolism on NPCs phenotype and function in NAFLD is unclear. The broad, long term goals of this project are to identify new mechanism by which fructose excess leads to the development of NAFLD, by uncovering metabolic pathways regulated by fructose that impact NPCs function. Preliminary data demonstrate that primary Kupffer cells and HSCs express KHK and fructose supplementation in vivo, increases tissue inhibitor of metalloproteinase (TIMP1) expression in Kupffer cells compared to diets supplemented with glucose or no monosaccharides in drinking water. In vitro, fructose treatment increases intracellular levels of acetate and formate in macrophages. Based on these observations, the specific aims of this project are: 1) to determine in vitro the mechanism by which fructose drives a profibrogenic phenotype in Kupffer cells and 2) characterize the phenotype and fructose metabolism in Kupffer cells under high fructose consumption during normal and NAFLD liver conditions. Our research design uses both in vivo and in vitro systems, to investigate the proposed aims. We integrate nuclear magnetic resonance and liquid chromatography mass spectrometry to trace 13C fructose in vitro (Aim 1) and in vivo (Aim 2) in Kupffer cells and to identify fructose generated metabolites that regulate fibrogenic gene and protein expression in NPCs. To investigate the role of TIMP-1 and acetate in Kupffer cells, small interfering RNA (siRNA) transfections (Aim 1) will be used in vitro. Data obtained from this proposal are expected to establish a foundation for new mechanisms of carbohydrate metabolism in NPCs and provide critical insight for the potential development of therapeutic targets for the prevention and treatment of NAFLD.

项目摘要 NAFLD在影响成年人和儿童的当今社会中逐渐变得越来越普遍。高的 果糖玉米糖浆的消耗与糖尿病，肥胖和非酒精性脂肪密切相关 肝病（NAFLD）。 NAFLD患者的果糖消耗量高两到三倍 与对照组相比，增加了小叶炎症和纤维化的存在。专业 小叶炎症和纤维化的贡献者是激活的肝非核细胞（NPC），， 由肝星状细胞（HSC）和库普弗细胞组成。果糖代谢一直是 主要在肝细胞中进行了研究，众所周知会诱导脂肪生成和氧化应激。这些 肝细胞中的事件已被证明会影响库普弗细胞功能，将细胞推向一个 炎症表型。然而，果糖代谢在NPCS表型和 NAFLD中的功能尚不清楚。该项目的长期目标是确定新机制 通过发现代谢途径，果糖过量导致NAFLD的发展 受影响NPC功能的果糖调节。初步数据证明了主要库普弗 细胞和HSC在体内表达KHK和补充果糖，增加了组织抑制剂 与补充葡萄糖的饮食相比，库普弗细胞中金属蛋白酶（TIMP1）的表达 或饮用水中没有单糖。在体外，果糖治疗增加了细胞内水平 巨噬细胞中的乙酸盐和甲酸盐。基于这些观察，该项目的具体目的 是：1）在体外确定果糖在其中驱动纤维化表型的机制 Kupffer细胞和2）表征高库普弗细胞中的表型和果糖代谢 正常和NAFLD肝脏条件下的果糖消耗。我们的研究设计在 体内和体外系统，以研究拟议的目的。我们整合了核磁共振 和液相色谱质谱法以在体外跟踪13C果糖（AIM 1）和体内（AIM 2） 在Kupffer细胞中，并鉴定果糖产生的代谢产生的代谢产物，这些代谢产生了调节纤维基因和蛋白质 在NPC中的表达。为了研究timp-1和乙酸盐在库普弗细胞中的作用，小干扰 RNA（siRNA）转染（AIM 1）将在体外使用。预计从该提案获得的数据是可以的 为NPC中的碳水化合物代谢的新机制建立基础 对预防和治疗NAFLD的治疗靶标的潜在发展的见解。