Circulating endocannabinoids in rats: Assay development and validation

大鼠循环内源性大麻素：检测方法开发和验证

• 批准号: 9306814
• 负责人: Cecilia J Hillard
• 金额: $ 7.7万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9306814
• 关键词: 2-arachidonylglycerol; Acute; Adipose tissue; Adult; Affect; American; Americas; Animal Model; Animals; Behavior; Behavioral; Biological Assay; Blood; Blood Circulation; Brain; CNR1 gene; Calories; Cardiovascular Diseases; Cessation of life; Circadian Rhythms; Comorbidity; Consumption; Data; Daughter; Detection; Diabetes Mellitus; Drug Addiction; Eating; Eating Disorders; Endocannabinoids; Energy Intake; Epidemic; Ethanolamines; Exertion; Exhibits; Face; Fatty acid glycerol esters; Food; Food Energy; Food Intake Regulation; Food deprivation (experimental); Fright; Future; Glucose; Glycogen; Health; High Prevalence; Human; Hyperphagia; Individual; Intake; Ions; Isotopes; Lead; Life Expectancy; Ligands; Light; Lipids; Malignant Neoplasms; Measurement; Measures; Metabolic; Modernization; Mood Disorders; Muscle; Nutrient; Obesity; Odors; Paracrine Communication; Pattern; Peripheral; Physiological; Plasma; Play; Pre-Clinical Model; Process; Psychological Stress; Public Health; Rattus; Receptor Signaling; Regulation; Relapse; Research Project Grants; Rewards; Role; Scheme; Series; Signaling Molecule; Site; Societies; Source; Stress; System; Testing; Thermogenesis; Time; Tissues; Translational Research; Translations; United States; Validation; Vitamins; Withdrawal; acute stress; assay development; awake; brain circulation; design; emotional factor; experimental study; falls; food consumption; food restriction; human data; innovation; interest; liquid chromatography mass spectrometry; nutrition; response; trend

The mechanisms that regulate the intake of food are vital for survival and health. While too little food consumption can lead to death, an equal or greater problem in 21st century America is the over-consumption of calorie-dense and inadequate consumption of vitamin-rich foods, resulting in obesity and poor nutrition. CB1 cannabinoid receptors (CB1R) in the brain play a vital role in the regulation of food intake, while activation of CB1R in peripheral tissues results in increased fat and glycogen storage; reduced glucose utilization by muscles; and reduced thermogenesis in brown adipose. Endocannabinoids are present in the human circulation and their concentrations are regulated by metabolic state and exhibit a circadian pattern. These previous findings lead us to the overall hypotheses that endocannabinoids in the circulation convey information about the need for additional caloric intake and that sustained, high endocannabinoid concentrations contribute to overeating. The objectives of this proposal are to establish and characterize an assay system in which endocannabinoid concentrations in the circulation of awake, freely behaving rats are determined. We will examine changes in endocannabinoid concentrations across the daily cycle; and following acute food restriction and stress, as these have all been shown to impact circulating endocannabinoid concentrations in humans. We will carry out three specific aims. The first aim is to optimize the endocannabinoid measurement in rat plasma; determining the limits of detection and quantification, and robustness. The second aim is to apply the assay to measure the circadian changes in circulating endocannabinoid concentrations in relationship to other metabolic signaling molecules; and to examine the effects of a brief period of food deprivation. The third aim will examine the time course of the effects of fear-evoking odor presentation on circulating endocannabinoids. Successful completion of these studies will provide the basis for future studies, including exploration of the mechanisms by which changes in energy status affect endocannabinoids in the circulation; the tissue source of the endocannabinoids in the circulation; and, most importantly, whether the endocannabinoids entering the brain from the circulation convey information about metabolic status to brain reward circuit activity. These studies are highly translatable to humans because endocannabinoid concentrations are easily measured in human blood. Indeed, the innovation of this project lies in its "beside to bench" translation of human observations to an animal model that is amenable to mechanistic studies. The studies in this project will increase our understanding of several processes important in the regulation of circulating endocannabinoid concentrations and will set the stage for future human and animal studies of their role in eating disorders, drug addiction and mood disorders.

调节食物摄入的机制对于生存和健康至关重要。虽然食物太少 消费会导致死亡，21世纪美国的一个同等甚至更大的问题是过度消费 热量密集和富含维生素的食物摄入不足，导致肥胖和营养不良。 CB1 大脑中的大麻素受体（CB1R）在调节食物摄入方面发挥着至关重要的作用，同时激活 外周组织中的 CB1R 导致脂肪和糖原储存增加；减少葡萄糖利用率 肌肉；并减少棕色脂肪的产热作用。内源性大麻素存在于人体中 循环及其浓度受代谢状态调节并表现出昼夜节律模式。这些 先前的发现使我们得出总体假设，即循环中的内源性大麻素传递信息 关于额外热量摄入的需要以及持续的高内源性大麻素浓度有助于 暴饮暴食。该提案的目标是建立并表征一个分析系统，其中 测定清醒、自由活动的大鼠循环中的内源性大麻素浓度。我们将 检查每日周期中内源性大麻素浓度的变化；和急性食物后 限制和压力，因为这些都已被证明会影响循环中的内源性大麻素浓度 人类。我们将实现三个具体目标。第一个目标是优化内源性大麻素的测量 大鼠血浆中；确定检测和定量的限度以及稳健性。第二个目标是 应用该测定来测量循环内源性大麻素浓度的昼夜变化 与其他代谢信号分子的关系；并检查短期食物的影响 剥夺。第三个目标将研究引起恐惧的气味呈现对人的影响的时间过程。 循环内源性大麻素。成功完成这些研究将为将来的学习奠定基础， 包括探索能量状态变化影响内源性大麻素的机制 循环;循环中内源性大麻素的组织来源；而且，最重要的是，是否 从循环进入大脑的内源性大麻素向大脑传递有关代谢状态的信息 奖励电路活动。这些研究高度适用于人类，因为内源性大麻素 人体血液中的浓度很容易测量。确实，这个项目的创新之处就在于它的“除了 “bench”将人类观察结果转化为适合机械研究的动物模型。 该项目的研究将增加我们对调节中重要的几个过程的理解 循环内源性大麻素浓度将为未来人类和动物研究奠定基础 在饮食失调、药物成瘾和情绪障碍中发挥作用。