Transcriptional Regulaton of Pro-Inflammatory Genes by Gambogic Acid

藤黄酸对促炎基因的转录调控

基本信息

批准号：
7686373
负责人：
KUMBLE SANDEEP PRABHU
金额：
$ 10.26万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-15 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7686373
关键词：
Acids Address Anti-Inflammatory Agents Anti-inflammatory Arachidonic Acids Asia Australia Biochemical Biological Biological Assay Biotin Bone Marrow Botanicals Carbon Chemicals Clusiaceae Complement Factor B Complementary Medicine Complementary and alternative medicine Confocal Microscopy DNA Footprint Development Disease Disease Progression Down-Regulation Enzymes Evaluation Family Figs - dietary Garcinia Gel Gene Expression Generations Genes Genetic Transcription Immune Immunoprecipitation In Vitro Inflammation Inflammatory Inflammatory Response Lipopolysaccharides Luciferases Methods Modeling Modification Molecular Molecular Mechanisms of Action Mus Nuclear Oxidation-Reduction Pathway interactions Personal Satisfaction Phosphotransferases Plant Resins Plasmids Play Polynesia Process Property Prostaglandin-Endoperoxide Synthase Proteins Public Health Recording of previous events Regulation Reporter Role Site Southern Africa Sulfhydryl Compounds TNFRSF5 gene Testing Therapeutic Traditional Medicine Transcriptional Regulation Tumor Necrosis Factor-alpha Tumor Necrosis Factors Western Blotting Xanthones adduct analog angiogenesis attenuation base carbonyl group chromatin immunoprecipitation cyclooxygenase 2 cytokine gambogic acid immunoregulation in vitro Model in vivo in vivo Model interest macrophage mutant p65 prevent promoter protein function response tool transcription factor tumor tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): This R21 application is submitted in response to an RFA (#AT-07-005) entitled "Mechanisms of Immune Modulation." The primary objective of our proposed studies is to understand the molecular mechanism of action of an important complementary medicine, gambogic acid (GA) that has a long history of medicinal use. GA is a polyprenylated xanthone, with an ?, ?-unsaturated carbonyl group, isolated from gamboges, the resin from Garcinia morella and Garcinia hanburyi. Many botanicals with such a reactive carbon center are known to possess interesting pharmacological properties. In this regard, GA, which possesses anti-tumor activity, is thought to possess anti-inflammatory activities that have so far remained untested. Due to the presence of the ?,?-unsaturated carbonyl group, GA is a potent Michael acceptor that can covalently interact with free thiol groups in proteins, including transcription factors. Our preliminary results demonstrate an inhibitory effect of GA on the bacterial lipopolysaccharide (LPS)-induced activation of nuclear factor-?B (NF-?B). NF-?B is a major transcription factor responsible for inflammation-driven disease progression. We will extend these studies to elucidate the underlying molecular mechanism of immune modulation of inflammation by GA with a specific focus on the transcription of two NF-?B target pro-inflammatory genes, cyclooxygenase (COX)-2 and tumor necrosis factor (TNF)-?. The specific hypothesis to be tested is that GA down-regulates the transcription of COX-2 and TNF? via the formation of covalent Michael adducts with protein components of the NF-?B cascade. The hypothesis will be tested using LPS-stimulated murine bone marrow-derived macrophages and validated in an in-vivo inflammation model, in C57/BL6 mice, in two Specific Aims: 1) To examine the effect of GA on the transcriptional regulation of COX-2 and TNF?, and 2) To elucidate the molecular mechanism of attenuation of NF-?B activation and function by GA. Studies will be focused on attenuation of inflammation by GA via the transcriptional regulation of COX-2 and TNF? and will involve the use of luciferase-based reporter and chromatin immunoprecipitation assays. Furthermore, the interaction of GA with protein components of the NF-?B pathway will be assessed by using biotinylated-GA in mass spectrometric and biochemical functional studies. Our long-term objective is to enhance the understanding of the molecular mechanism of action of botanicals used in complementary and alternative medicine for their role in immune regulation of inflammation. PUBLIC HEALTH REVELANCE: Gamboges, the resin from Garcinia morella and G. hanburyi that belong to the family Clusiaceae (Guttiferae), which are native to Asia, Australia, Southern Africa, and Polynesia, are rich in a polyprenylated xanthone, gambogic acid (GA). There is a long history of medicinal use of Garcinia extracts against many ailments. Interestingly, the anti-tumor activity of GA has been well demonstrated and is thought to arise partly due to the anti-inflammatory activity associated with GA. However, the anti-inflammatory activity of GA has not been tested. We propose to study the molecular basis of the down-regulation of NF-?B-dependent expression of pro-inflammatory genes by GA in in vitro and in vivo models of inflammation.

描述（由申请人提供）：此 R21 申请是为了响应题为“免疫调节机制”的 RFA (#AT-07-005) 而提交的。我们提出的研究的主要目的是了解一种重要的补充药物——具有悠久药用历史的藤黄酸（GA）的分子作用机制。 GA 是一种聚异戊二烯化呫吨酮，具有 α,β-不饱和羰基，从藤黄中分离出来，藤黄是藤黄和汉伯里藤黄的树脂。已知许多具有这种活性碳中心的植物药具有有趣的药理学特性。在这方面，具有抗肿瘤活性的GA被认为具有迄今为止尚未经过测试的抗炎活性。由于α,β-不饱和羰基的存在，GA 是一种有效的迈克尔受体，可以与蛋白质（包括转录因子）中的游离硫醇基团共价相互作用。我们的初步结果表明，GA 对细菌脂多糖 (LPS) 诱导的核因子-κB (NF-κB) 激活具有抑制作用。 NF-κB 是负责炎症驱动的疾病进展的主要转录因子。我们将扩展这些研究，以阐明 GA 免疫调节炎症的潜在分子机制，特别关注两个 NF-κB 靶促炎基因、环氧合酶 (COX)-2 和肿瘤坏死因子 (TNF) 的转录-？。待检验的具体假设是GA下调COX-2和TNF的转录？通过与 NF-κB 级联的蛋白质成分形成共价迈克尔加合物。该假设将使用 LPS 刺激的小鼠骨髓来源的巨噬细胞进行测试，并在 C57/BL6 小鼠的体内炎症模型中进行验证，有两个具体目标：1) 检查 GA 对 COX 转录调节的影响-2 和 TNF?，以及 2) 阐明 GA 减弱 NF-κB 激活和功能的分子机制。研究将集中于 GA 通过 COX-2 和 TNF 的转录调节来减轻炎症？并将涉及使用基于荧光素酶的报告基因和染色质免疫沉淀测定。此外，将通过在质谱和生化功能研究中使用生物素化-GA来评估GA与NF-κB途径的蛋白质成分的相互作用。我们的长期目标是加强对补充和替代医学中植物药在炎症免疫调节中作用的分子作用机制的了解。公众健康启示：藤黄是来自藤黄果（Garcinia morella）和藤黄藤（G. hanburyi）的树脂，属于藤黄科（Guttiferae），原产于亚洲、澳大利亚、南非和波利尼西亚，富含聚异戊二烯化氧杂蒽酮、藤黄酸（GA ）。藤黄果提取物用于治疗多种疾病有着悠久的历史。有趣的是，GA 的抗肿瘤活性已得到充分证明，并且被认为部分归因于 GA 相关的抗炎活性。然而，GA 的抗炎活性尚未经过测试。我们建议在体外和体内炎症模型中研究 GA 下调 NF-κB 依赖性促炎基因表达的分子基础。