ANTI INFLAMMATORY AND ANTIFIBROTIC ACTIONS OF THYMOSIN BETA 4 IN ALD

胸腺肽 4 在 ALD 中的抗炎和抗纤维化作用

基本信息

批准号：
8854003
负责人：
RAJ M LAKSHMAN
金额：
$ 17.57万
依托单位：
GEORGE WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8854003
关键词：
ADD-1 protein Acetaldehyde Aftercare Alcoholic Liver Diseases Alcohols Aldehydes Anti-Inflammatory Agents Anti-inflammatory Appearance Applications Grants Architecture Biochemical Biochemistry Chemistry Chronic Clinical Treatment Clinical Trials Collagen Collagen Gene Dissociation Down-Regulation Endotoxins Epigenetic Process Ethanol Extracellular Matrix Proteins Eye Fetoprotein Fibronectins Fibrosis Future Gene Proteins Generations Genes Goals Health Heart Hepatic Hepatic Fibrogenesis Hepatic Stellate Cell Hepatic Tissue Hepatocyte In Vitro Inflammation Inflammatory Injury Interleukin-1 beta Lead Lipids Liver Liver diseases Mediating Metabolic Methyl-CpG-Binding Protein 2 Modeling Molecular Molecular Biology Mus Myofibroblast Natural regeneration Oxidative Stress PPAR gamma Peptides Pericytes Peroxisome Proliferator-Activated Receptors Phenotype Phosphorylation Platelet-Derived Growth Factor Platelet-Derived Growth Factor beta Receptor Process Property Proteins Reactive Oxygen Species Reporting Role Serum Serum Markers Signal Transduction Site Skin Smooth Muscle Actin Staining Method Techniques Testing Therapeutic Agents Thymosin Time Tissues Tumor Necrosis Factor-alpha Up-Regulation Vitamin A adduct base cytokine fibrogenesis in vitro Model in vivo innovation liver inflammation liver injury mouse model novel novel therapeutics overexpression preclinical study prevent receptor stellate cell thymosin beta(4)transdifferentiation

项目摘要

DESCRIPTION (provided by applicant): Based on the well accepted two-hit model for ALD, Chronic Ethanol (EtOH)/LPS generated endotoxins activate NFκB that up regulates TNFα, and IL1ß the potent proinflammatory cytokines causing hepatocyte injury. Further, EtOH induced Cyp2E1 and ADH lead to metabolic generation of acetaldehyde and increased reactive oxygen species (ROS), which activate quiescent HSC to myofibroblasts resulting in up regulation of fibrogenic genes, platelet derived growth factor ß-receptor (PDGFßr), α-smooth muscle actin (αSMA) and extracellular matrix proteins (ECM), collagen I, III, and fibronectin and epigenetic repressor gene, methyl-CpG binding protein 2 (MeCP2). In contrast, adipogenic genes, peroxisome proliferator-activated receptor γ (PPARγ), and sterol regulatory element-binding protein 1c (SREBP1c) are suppressed resulting in the loss of their vitamin A stores and their transdifferentiation from quiescent lipid storing phenotype to active myofibroblastic phenotype. Significantly, thymosin ß4 (Tß4), a bioactive peptide, is reported to prevent inflammation and fibrosis in many extra-hepatic tissues. Based on these, PI hypothesizes that Tß4's anti-inflammatory and anti-fibrogenic actions against EtOH/LPS liver injury are mediated by (i) inhibiting the activation of NFκB by blocking the phosphorylation and dissociation of IκB and thereby prevent the up regulation of TNFα, and IL1ß the potent proinflammatory cytokines and consequent liver injury, and (ii) suppressing the up regulated MeCP2, that coordinately reverses (a) the down regulated adipogenic genes and (b) up regulated fibrogenic genes and thereby prevent the trans-differentiation of HSC from lipid-storing pericytes to myofibroblasts. We also hypothesize that Tß4 elicits its above actions by overexpressing miR132 that suppresses MeCP2 overexpression caused by EtOH/LPS. PI has the following encouraging preliminary results in the EtOH/LPS mouse model to reassure the feasibilities of his novel exploratory approaches: Tß4 protects against EtOH/LPS induced 1. Up-regulation of NFκB signaling cascade, pIκB, TNFα, IL1ß and consequent serum markers for liver injury; 2. up regulation of hepatic MeCP2, PDGFßr, αSMA, Col1α1 & proteins; and 3. down-regulation of adipogenic gene, PPARγ. As a result, we propose that Tß4 blocks the (i) activation of NFκB, TNFα and inflammatory cascade and (ii) transdifferentiation of quiescent HSC to fibrogenic HSC; yet, Tß4 maintains hepatocyte regeneration. Thus, the major goals of our innovative proposal are to accomplish the following specific aims: Specific Aim 1. What are the possible mechanism/s of action/s of Tß4 "Pre- and Post-treatment to protect/alleviate" EtOH/LPS-mediated up regulation of NFκB signaling cascade, TNFα & IL1ß, and consequent serum and liver markers for liver injury? Specific Aim 2: What are the possible mechanism/s of action/s of Tß4 "Pre- and Post-treatment to protect/alleviate" against EtOH/LPS-mediated (a) up regulation of hepatic fibrogenic genes and their products? and (b) down regulation of adipogenic genes and their products? Are there corresponding morphological changes in liver cell architecture as well as in HSC phenotype? PI plans to approach using established EtOH/LPS two-hit mouse in vivo & in vitro model utilizing biochemistry, molecular biology, immuno- & histo-chemistry techniques. PI has a strong molecular biology and biochemical group that has the expertise in all aspects of this proposal. This may lead to novel therapy for ALD.

描述（由适用提供）：基于ALD的公认的两次打击模型，慢性乙醇（ETOH）/LPS生成的内毒素激活了NFκB，从而增强了TNFα，而IL1ß则有效的促炎细胞因子会导致肝细胞损伤。 Further, EtOH induced Cyp2E1 and ADH lead to metabolic generation of acetaldehyde and increased reactive oxygen species (ROS), which activate quiescent HSC to Myofibroblasts resulting in up regulation of fibrogenic genes, platelet derived growth factor ß-receptor (PDGFßr), α-smooth muscle actin (αSMA) and extracellular matrix proteins （ECM），胶原I，III和纤连蛋白和表观遗传反射剂基因，甲基-CPG结合蛋白2（MECP2）。相反，抑制脂肪生成基因，过氧化物体增殖物激活的受体γ（PPARγ）和固醇调节元件结合蛋白1C（SREBP1C）导致其维生素A储备的损失及其因静止的脂质脂质储存的现象型造型型肌爆发型造成的维生素A储备及其跨差异。值得注意的是，据报道，胸腺素ß4（Tß4）是一种生物活性肽，可防止许多肝外组织的感染和纤维化。 Based on these, PI hypothesizes that Tß4's anti-inflammatory and anti-fibrogenic actions against EtOH/LPS liver injury are mediated by (i) inhibiting the activation of NFκB by blocking the phosphorylation and dissociation of IκB and thereby prevent the up regulation of TNFα, and IL1ß the potent proinflammatory cytokines and consequent liver injury, and (ii) suppressing向上调节的MECP2，该MECP2协同逆转（a）下调的脂肪生成基因和（b）提高调节的纤维基因基因，从而防止HSC从脂质周围的周细胞转移到肌纤维细胞。我们还假设Tß4通过过表达MiR132抑制了由EtOH/LPS引起的MECP2过表达来引起上述作用。 PI在ETOH/LPS小鼠模型中具有以下令人鼓舞的初步结果，可以解决他的新型探索方法的可行性：Tß4可以防止ETOH/LPS诱导的1。NFκB信号级联，PIκB，TNFα，TNFα，IL1ß和结果型血清标记对肝损伤的上调； 2。肝MECP2，PDGFßR，αSMA，COL1α1和蛋白质的调节；和3。掺杂基因PPARγ的下调。结果，我们提出Tß4阻止了NFκB，TNFα和炎症级联反应的（i）激活，以及（ii）将静态HSC转变为纤维基因HSC；然而，Tß4保持肝细胞再生。这是我们创新提案的主要目标是实现以下特定目的：特定目标1。具体目标2：Tß4“预防/减轻eTOH/LPS介导的（a）调节肝纤维基因及其产物的ETOH/LPS介导的（a）调节的Tß4“可保护/减轻”的作用/s的可能机理/s是什么？（b）减少掺杂基因及其产物的调节？肝细胞结构以及HSC表型中是否存在相应的形态变化？ PI计划利用生物化学，分子生物学，免疫和组织化学技术在体内和体外模型中使用已建立的ETOH/LPS两次小鼠进行接近。 PI具有强大的分子生物学和生化群，在该提案的各个方面都有专业知识。这可能会导致对ALD的新疗法。