Novel Mechanisms of NSAID-induced GI Toxicity

NSAID 引起胃肠道毒性的新机制

基本信息

批准号：
8232604
负责人：
JAMES D LILLICH
金额：
$ 33.3万
依托单位：
KANSAS STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8232604
关键词：
Accounting Address Adhesions Adverse effects Affect Anti-Inflammatory Agents Anti-inflammatory Binding Bromodeoxyuridine Calpain Cardiovascular system Cell Adhesion Cell Migration Inhibition function Cell Proliferation Cell membrane Cells Complex Confocal Microscopy Data Databases Development Drug toxicity Drug usage EGF Signaling Pathway EGF gene Enterocytes Enzymes Epidermal Growth Factor Epidermal Growth Factor Receptor Epithelial Epithelial Cells Equilibrium Fluorescence Resonance Energy Transfer Future Gastrointestinal tract structure Glean Goals Healed Hospitalization Immunoblotting Immunofluorescence Immunologic In Vitro Inflammation Intestines Kinetics Label Laboratories Life Link Measures Medical Membrane Membrane Potentials Messenger RNA Migration Assay Mitogen-Activated Protein Kinase 3 Molecular Non-Steroidal Anti-Inflammatory Agents PTEN gene Pain Pain management Pathway interactions Patients Peptide Hydrolases Pharmaceutical Preparations Pharmacotherapy Phosphatidylinositol 4,5-Diphosphate Phosphorylation Physiologic pulse Procedures Process Production Prostaglandin-Endoperoxide Synthase Protein Biosynthesis Proteins Proteolysis Rattus Receptor Activation Relative (related person)Research Risk Safety Signal Pathway Specificity Talin Tissues Toxic effect Ulcer Villus Voltage-Gated Potassium Channel Work Wound Healing base cell motility cellular imaging crypt cell cyclooxygenase 1 cyclooxygenase 2 design drug development gastrointestinal gastrointestinal epithelium healing improved in vivo innovation inorganic phosphate insight medical complication membrane activity migration novel phosphoinositide-3,4,5-triphosphate protein expression receptor repaired research study wound

项目摘要

DESCRIPTION (provided by applicant): Nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most widely used drugs worldwide for the relief of inflammation and pain. However, NSAIDs cause adverse gastrointestinal (GI) side-effects in a large proportion of people who use them, many of whom require hospitalization to recover. Traditionally, NSAID- induced GI toxicity was associated with the inhibition of cyclooxygenase (COX) enzymes, particularly COX-1. Recent evidence however, indicates that COX inhibition does not account for the full range of toxic effects observed in the GI tract. Additionally, the discovery of the adverse cardiovascular effects of COX-2 specific NSAIDs has demonstrated the importance of identifying mechanisms through which NSAIDs cause GI toxicity. Accordingly, we propose to investigate the molecular mechanisms underlying inhibition of restitution by NSAIDs, one of the negative effects that NSAIDs have on GI epithelia. GI epithelia form a barrier that separates the internal milieu from the harsh conditions of the lumen of the GI. Restitution is the cell proliferation-free process by which wounds or gaps in the epithelial barrier are repaired by the spreading and migration of neighboring cells into the wound. NSAIDs with ulcerogenic potential inhibit cell migration in intestinal epithelial cells. Our research group has shown that this inhibition is associated with a decrease in the expression and activity of calpain proteases and depolarization of membrane potential (Em). On the basis of these data, we hypothesize that NSAIDs impair intestinal epithelial cell migration and inhibit calpain activity through a signaling pathway that is highly sensitive to drug-induced changes in Em. Therefore, in Specific Aim 1 we will determine if NSAIDs impair de-adhesion of cell contacts by measuring the number and distribution of cellular adhesions as well as the expression and rate of turnover of specific adhesion components. In Specific Aim 2, we will determine if NSAIDs or Em depolarization modulate calpain activity through disruption of EGF-stimulated phosphorylation of calpains or disruption of PIP balance by measuring expression and activation of the EGF pathway, PIP levels, and calpain activity and localization to the membrane. In Specific Aim 3, we will extend our in vitro experiments by determining the effects of NSAIDs on cell migration, calpain and Kv channel expression, and calpain participation in membrane localized complexes in rats treated with NSAIDs. These innovative experiments have the potential to identify molecular mechanisms through which NSAIDs decrease calpain activity, in addition to determining the effects of reductions in calpain activity on cellular de-adhesion during migration both in vitro and in vivo. This project will provide basic information on wound healing processes in intestinal cells that are applicable to other tissues, document the effects of NSAIDs on these processes, and give insight into how NSAIDs may affect the healing process in other tissues. The research also has great potential for aiding in the development of new drugs in this class capable of ameliorating pain without the adverse side effects of current drugs. PUBLIC HEALTH RELEVANCE: The widespread use of nonsteroidal anti-inflammatory drugs (NSAIDs) for the alleviation of inflammation and pain poses a major medical risk to patients due to the common toxic effects these drugs have on the gastrointestinal tract. Our research is designed to elucidate novel mechanisms through which these toxic effects occur in order to eliminate the side effects associated with NSAID use and enhance the safety of future drugs of this class. This research may help reduce or eliminate the loss of billions of dollars per year due to medical complications resulting from NSAID-induced GI toxicity.

描述（由申请人提供）：非甾体类抗炎药（NSAID）是全世界用于缓解炎症和疼痛最广泛使用的药物之一。然而，非甾体抗炎药会对很大一部分使用者产生不良的胃肠道 (GI) 副作用，其中许多人需要住院才能康复。传统上，NSAID 引起的胃肠道毒性与环氧合酶 (COX)，特别是 COX-1 的抑制有关。然而，最近的证据表明，COX 抑制并不能解释胃肠道中观察到的全部毒性作用。此外，COX-2 特异性非甾体抗炎药对心血管的不良影响的发现证明了确定非甾体抗炎药引起胃肠道毒性机制的重要性。因此，我们建议研究 NSAID 抑制恢复的分子机制，这是 NSAID 对胃肠道上皮细胞的负面影响之一。胃肠道上皮形成一道屏障，将内部环境与胃肠道管腔的恶劣条件分开。恢复是一种无细胞增殖的过程，通过邻近细胞扩散和迁移到伤口中来修复上皮屏障中的伤口或间隙。具有致溃疡潜力的非甾体抗炎药抑制肠上皮细胞中的细胞迁移。我们的研究小组表明，这种抑制与钙蛋白酶表达和活性的降低以及膜电位 (Em) 的去极化有关。基于这些数据，我们假设 NSAIDs 通过对药物诱导的 Em 变化高度敏感的信号通路损害肠上皮细胞迁移并抑制钙蛋白酶活性。因此，在具体目标 1 中，我们将通过测量细胞粘附的数量和分布以及特定粘附成分的表达和周转率来确定 NSAID 是否会损害细胞接触的脱粘附。在具体目标 2 中，我们将通过测量 EGF 通路的表达和激活、PIP 水平以及钙蛋白酶活性和定位来确定 NSAID 或 Em 去极化是否通过破坏 EGF 刺激的钙蛋白酶磷酸化或破坏 PIP 平衡来调节钙蛋白酶活性。膜。在具体目标 3 中，我们将通过确定 NSAID 对细胞迁移、钙蛋白酶和 Kv 通道表达以及钙蛋白酶参与 NSAID 治疗大鼠膜定位复合物的影响来扩展我们的体外实验。这些创新实验除了确定钙蛋白酶活性降低对体外和体内迁移过程中细胞脱粘附的影响外，还有可能确定非甾体抗炎药降低钙蛋白酶活性的分子机制。该项目将提供适用于其他组织的肠细胞伤口愈合过程的基本信息，记录非甾体抗炎药对这些过程的影响，并深入了解非甾体抗炎药如何影响其他组织的愈合过程。该研究还具有帮助开发此类新药的巨大潜力，这些新药能够减轻疼痛，且没有现有药物的不良副作用。公共卫生相关性：广泛使用非甾体类抗炎药 (NSAID) 来缓解炎症和疼痛，由于这些药物对胃肠道具有常见的毒性作用，因此给患者带来了重大医疗风险。我们的研究旨在阐明这些毒性作用发生的新机制，以消除与使用 NSAID 相关的副作用并提高未来此类药物的安全性。这项研究可能有助于减少或消除每年因非甾体抗炎药引起的胃肠道毒性引起的医疗并发症而造成的数十亿美元的损失。