Determining the Role of FAAH and MAGL in the Modulation of Mechanical Allodynia

确定 FAAH 和 MAGL 在调节机械异常性疼痛中的作用

• 批准号: 7812802
• 负责人: Lamont Booker
• 金额: $ 3.8万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-25 至 2011-05-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7812802
• 关键词: 2-arachidonylglycerol; Absence of pain sensation; Address; Agonist; American; Analgesics; Anti-Inflammatory Agents; Anti-inflammatory; Area; Attention; Attenuated; Behavior; Binding; Bradykinin; CNR2 gene; Cannabinoids; Carrageenan; Cell Wall; Cells; Development; Dinoprostone; Disease; Edema; Endocannabinoids; Endotoxins; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzyme-Linked Immunosorbent Assay; Enzymes; Fibromyalgia; Filament; Freund' s Adjuvant; Genetically Modified Animals; Goals; Gram-Negative Bacteria; Hyperalgesia; Immune; Inflammation; Inflammatory; Inflammatory Response; Injection of therapeutic agent; Interleukin-6; Knockout Mice; Life; Life Style; Lipopolysaccharides; Marijuana; Mechanics; Mediating; Mediator of activation protein; Modeling; Monoacylglycerol Lipases; Motor Activity; Mus; Neuraxis; Neuropathy; Nociception; Nociceptors; Operative Surgical Procedures; Pain; Pharmaceutical Preparations; Play; Postherpetic neuralgia; Postoperative Period; Property; Proteins; Role; Signal Transduction; Societies; Stimulus; Substance P; System; Tail; Techniques; Technology; Testing; Therapeutic; Time; Wild Type Mouse; allodynia; anandamide; biological systems; cannabinoid receptor; chemokine; combat; cytokine; fatty acid amide hydrolase; in vivo; inflammatory pain; inhibitor/antagonist; mechanical allodynia; receptor; response; tool

DESCRIPTION (provided by applicant): Pain of any type is typically the underlying result of many disorders and diseases which compromises millions of Americans lives and lifestyles according to the American Pain Society. More specifically allodynia, which is a painful response to non-noxious stimuli is gaining more attention as it is commonly associated with disorders such as neuropathy, postherpetic neuralgia, fibromyalgia, and post operative surgeries. However, treatments for allodynia are not always effective due to the abundance of mediators contributing to its occurrence. Exogenously administered cannabinoids are capable of binding to the cannabinoid receptors, which then signals a downstream cascade to occur, and ultimately produce analgesia. Similarily, endocannabinoids (anandamide; AEA and 2-AG) also bind to the cannabinoid receptors; however, their activation at the receptor level is tremendously decreased due to degradative enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL), which metabolizes AEA and 2-AG respectively. The goal of this project is to investigate the roles of FAAH and MAGL in modulating mechanical allodynia. Allodynia can be induced by intraplantar injections of low concentrations of lipopolysaccharide, sustainable up to 48 hrs and tested using calibrated von frey filaments. To elucidate FAAH and MAGL roles in vivo, we will combine the use of pharmacological tools (selective receptor and enzyme inhibitors) and genetically modified animals (receptor, enzyme, and conditional knockout mice). Additionally, we will quantify endogenous cannabinoid levels using LC/MS/MS technology for regional specific areas that are important in modulating mechanical allodynia. In addition to lipopolysaccharide directly inducing allodynia, a secondary response to the endotoxin results in the release of cytokines. Therefore, since a final goal of this project is to investigate the relevance of FAAH and MAGL in altering in vivo cytokine levels to determine if they also contribute to the allodynic response. We will use ELISA techniques to quantify actual protein levels of both pro-inflammatory and anti-inflammatory cytokines produced. These studies will correlate the importance of FAAH and MAGL in regulating pain and inflammation. Additionally, it will further the understanding of the endocannabinoid system and its potential implications in therapeutic treatments.

描述（由申请人提供）：根据美国疼痛协会的说法，任何类型的疼痛通常是许多疾病和疾病的潜在结果，这些疾病和疾病危及数百万美国人的生活和生活方式。更具体地说，异常性疼痛是对非有害刺激的疼痛反应，它正受到越来越多的关注，因为它通常与神经病变、带状疱疹后神经痛、纤维肌痛和术后手术等疾病有关。然而，异常性疼痛的治疗并不总是有效的，因为大量的介质有助于其发生。外源性给予的大麻素能够与大麻素受体结合，然后向下游级联发出信号，并最终产生镇痛作用。类似地，内源性大麻素（大麻素; AEA和2-AG）也与大麻素受体结合;然而，由于降解酶脂肪酸酰胺水解酶（FAAH）和单酰基甘油脂肪酶（MAGL），它们在受体水平上的活化显著降低，所述降解酶分别代谢AEA和2-AG。本项目的目的是研究FAAH和MAGL在调节机械性异常性疼痛中的作用。异常性疼痛可以通过足底注射低浓度的脂多糖来诱导，持续长达48小时，并使用校准的von frey细丝进行测试。为了阐明FAAH和MAGL在体内的作用，我们将联合收割机结合使用药理学工具（选择性受体和酶抑制剂）和转基因动物（受体，酶和条件敲除小鼠）。此外，我们将使用LC/MS/MS技术量化内源性大麻素水平，用于调节机械性异常性疼痛的重要区域。除了脂多糖直接诱导异常性疼痛外，对内毒素的继发性反应导致细胞因子的释放。因此，由于本项目的最终目标是研究FAAH和MAGL在改变体内细胞因子水平方面的相关性，以确定它们是否也有助于异常性疼痛反应。我们将使用ELISA技术来定量产生的促炎和抗炎细胞因子的实际蛋白水平。这些研究将使FAAH和MAGL在调节疼痛和炎症中的重要性相互关联。此外，它将进一步了解内源性大麻素系统及其在治疗中的潜在影响。