Mechanisms of Probiotics in Alcoholic Liver Disease

益生菌治疗酒精性肝病的机制

• 批准号: 9766984
• 负责人: WENKE FENG
• 金额: $ 30.8万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-15 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9766984
• 关键词: Acute; Alcohol abuse; Alcohol consumption; Alcohol-Induced Disorders; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcohols; Animal Model; Attenuated; Bacteria; Bacterial Translocation; Butyric Acids; Carbohydrates; Cell Line; Chronic; Cirrhosis; Data; Development; Disease Progression; Effectiveness; Endotoxins; Enzymes; Epithelial; Epithelial Cells; Etiology; Experimental Models; Fatty Liver; Fibrosis; Functional disorder; Goals; Goblet Cells; Health; Hepatic; Histology; Histone Deacetylase; Homeostasis; Human; Hypoxia Inducible Factor; Immune response; Impairment; In Vitro; Inflammation; Injury; Intervention; Intestinal Diseases; Intestinal permeability; Intestines; Knockout Mice; Laboratories; Lactobacillus casei rhamnosus; Leaky Gut; Liver; Mediating; Metagenomics; Morbidity - disease rate; Mucous Membrane; Mucous body substance; Mus; Oral Administration; Outcome; Pathologic; Patients; Pilot Projects; Plasma; Play; Preparation; Prevention strategy; Prevention therapy; Probiotics; Process; Production; Proteins; Recombinants; Regulation; Reporting; Role; Sampling; Signal Transduction; Steatohepatitis; Supplementation; Testing; Tight Junctions; Volatile Fatty Acids; alcohol exposure; antimicrobial peptide; bacterial metabolism; base; cathelicidin; cathelicidin antimicrobial peptide; comparative efficacy; compare effectiveness; dysbiosis; gastrointestinal epithelium; gut microbiota; improved; in vivo; lipid metabolism; liver inflammation; liver injury; metabolomics; mortality; mouse model; novel therapeutic intervention; occludin; prevent; protective effect; public health relevance; repaired; transcription factor; trefoil factor; Acute; Alcohol abuse; Alcohol consumption; Alcohol-Induced Disorders; Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcohols; Animal Model; Attenuated; Bacteria; Bacterial Translocation; Butyric Acids; Carbohydrates; Cell Line; Chronic; Cirrhosis; Data; Development; Disease Progression; Effectiveness; Endotoxins; Enzymes; Epithelial; Epithelial Cells; Etiology; Experimental Models; Fatty Liver; Fibrosis; Functional disorder; Goals; Goblet Cells; Health; Hepatic; Histology; Histone Deacetylase; Homeostasis; Human; Hypoxia Inducible Factor; Immune response; Impairment; In Vitro; Inflammation; Injury; Intervention; Intestinal Diseases; Intestinal permeability; Intestines; Knockout Mice; Laboratories; Lactobacillus casei rhamnosus; Leaky Gut; Liver; Mediating; Metagenomics; Morbidity - disease rate; Mucous Membrane; Mucous body substance; Mus; Oral Administration; Outcome; Pathologic; Patients; Pilot Projects; Plasma; Play; Preparation; Prevention strategy; Prevention therapy; Probiotics; Process; Production; Proteins; Recombinants; Regulation; Reporting; Role; Sampling; Signal Transduction; Steatohepatitis; Supplementation; Testing; Tight Junctions; Volatile Fatty Acids; alcohol exposure; antimicrobial peptide; bacterial metabolism; base; cathelicidin; cathelicidin antimicrobial peptide; comparative efficacy; compare effectiveness; dysbiosis; gastrointestinal epithelium; gut microbiota; improved; in vivo; lipid metabolism; liver inflammation; liver injury; metabolomics; mortality; mouse model; novel therapeutic intervention; occludin; prevent; protective effect; public health relevance; repaired; transcription factor; trefoil factor

 DESCRIPTION (provided by applicant): Severe alcoholic liver disease (ALD) has a high morbidity and mortality. Recent studies demonstrated that probiotics reversed alcohol-induced hepatic steatosis and inflammation, and improved liver enzymes in animal models and in patients. Our laboratory showed that administration of a probiotic strain, Lactobacillus rhamnosus Gorbach-Goldin (LGG), significantly improved liver enzymes and histology in alcohol treated mice. However, the beneficial mechanisms of action of LGG, either in viable bacteria form or in culture supernatant form, are not clear. Our overall hypothesis is that LGG increases intestinal cathelicidin-related antimicrobial peptide (CRAMP), intestinal trefoil factor (ITF) expression and intestinal short chain fatty acids (SCFAs), leading to improved gut microbiota homeostasis, intestinal barrier integrity and hepatic fat metabolism, and these are likely the mechanisms by which LGG supplementation attenuates alcohol induced liver injury. We evaluate mechanisms of LGG action with 4 specific aims: Aim 1 will determine the role of CRAMP in the alcohol-mediated changes in gut microbiota and the role of CRAMP in the beneficial effects of LGG in experimental ALD. We will use CRAMP knockout mouse model of ALD to determine the role of CRAMP and LGG in alcohol-induced changes in gut microbiota using a metagenomic approach. Aim 2 will determine the role of LGG in promoting ITF expression and evaluate whether ITF positively modulates intestinal tight junctions as a mechanism for improved intestinal barrier integrity in experimental ALD. We will use an ITF-secreting goblet cell line to evaluate the role of LGG in the production of ITF and the role of ITF in the modulation of intestinal tight junctions in an epithelial cell line. We will also examine th strategy that oral administration of recombinant human ITF attenuates alcohol-induced intestinal barrier and liver injury. Aim 3 will determine the role and mechanisms of butyric acid in the beneficial effects of LGG on intestinal tight junctions in experimental ALD. A metabolomics approach will be used to analyze the changes in SCFAs induced by alcohol and LGG intervention in the intestinal lumen. We will explore the potential mechanisms underlying the role of butyric acid in the regulation of occludin expression involving intestinal histone deacetylases (HDACs), miR122a, and HIF--mediated signaling in vitro and in vivo. Aim 4 will evaluate and compare the effectiveness of LGG supernatant (LGGs) with viable LGG (vLGG) and heat-inactivated LGG (hiLGG) in the prevention/therapy of ALD in mouse models. These three LGG preparations will be tested for the effectiveness in the prevention/therapy of ALD in three experimental models of ALD of steatosis, steatohepatitis and fibrosis. This study will have a major impact on the development of probiotic-based new therapeutic strategy for the prevention and treatment of ALD.

 描述（由适用提供）：严重的酒精性肝病（ALD）具有高发病率和死亡率。最近的研究表明，益生菌逆转了酒精引起的肝脂肪变性和注射，并改善了动物模型和患者的肝酶。我们的实验室表明，给予益生菌菌株，鼠尾草乳杆菌gorbach---------- gordin（LGG），可显着改善酒精治疗的小鼠的肝酶和组织学。然而，尚不清楚以可行的细菌形式或培养上清液形式的LGG作用的有益机理。 Our overall hypothesis is that LGG increases intestinal cathelicidin-related antimicrobial peptide (CRAMP), intestinal trefoil factor (ITF) expression and intestinal short chain fatty acids (SCFAs), leading to improved gut microbiota homeostasis, intestinal barrier integrity and hepatitic fat Metabolism, and these are likely the mechanisms by which LGG supplementation attenuates酒精引起的肝损伤。我们以4个特定目的评估LGG作用的机制：AIM 1将确定抽筋在肠道菌群中的肠道变化中的作用，以及抽筋在LGG在实验ALD中的有益作用中的作用。我们将使用ALD的抽筋敲除小鼠模型来确定使用元基因组方法在酒精诱导的肠道菌群变化中痉挛和LGG的作用。 AIM 2将确定LGG在促进ITF表达中的作用，并评估ITF是否积极调节肠道紧密连接，作为改善实验ALD肠道屏障完整性的机制。我们将使用分泌ITF的杯状细胞系来评估LGG在ITF生产中的作用和ITF的作用 在上皮细胞系中肠紧密连接的调节中。我们还将研究策略，口服重组人ITF会减轻酒精引起的肠道屏障和肝损伤。 AIM 3将确定丁酸在LGG对实验ALD肠紧密连接的有益作用中的作用和机制。代谢组学方法将用于分析肠道内酒精和LGG干预引起的SCFA的变化。我们将探讨丁酸在调节涉及肠道组蛋白脱乙酰基酶（HDACS），miR122a和HIF-介导的信号传导的体外和体内的咬合蛋白表达中的作用的潜在机制。 AIM 4将评估和比较LGG上清液（LGGS）的有效性（LGG）和可行的LGG（VLGG）和热灭活的LGG（Hilgg）在小鼠模型中ALD的预防/治疗中的有效性。这三种LGG制剂将在三种ALD的脂肪变性，脂肪性肝炎和纤维化的ALD实验模型中进行预防/ALD治疗的有效性。这项研究将对基于益生菌的预防和治疗ALD的新治疗策略的发展产生重大影响。