ANANDAMIDE/THC INDUCED ARACHIDONIC ACID RELEASE

花生四烯酸/四氢大麻酚引起的花生四烯酸释放

• 批准号: 2668143
• 负责人: SUMNER HOWARD BURSTEIN
• 金额: $ 16.06万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-03-15 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2668143
• 关键词: anandamide; arachidonate; biological signal transduction; cannabinoid receptor; cannabinoids; eicosanoid metabolism; eicosanoids; mitogen activated protein kinase; phospholipase A2; phospholipase D; protein kinase C; receptor binding; tissue /cell culture

The goal of the experiments proposed in this application is to determine the biochemical events that lead to cannabinoid-induced arachidonic acid release. The studies to be carried out will be focused on the signal transduction processes following cannabinoid binding to its receptor. The question of receptor mediation in this process will be addressed by the use of anti sense oligodeoxynucleotides in addition to the more conventional ligand binding approach. Oligos will be designed to reduce, or eliminate completely, the expression of the cannabinoid receptor genes and compared with subsequent changes in arachidonic acid release. The term cannabinoid is meant to include both exogenous substances, such as THC, and endogenous molecules like anandamide. The systems to be used for these investigations will be isolated, intact cells such as mouse peritoneal macrophages, neuroblastoma and Chinese-hamster ovary lines. The experiments are aimed at defining roles for phospholipase D, protein kinase C, MAP kinase and phospholipase A2 in mediating cannabinoid- stimulated release of arachidonic acid and the subsequent production of eicosanoids. Direct measurement of enzyme activities will be made over a range of cannabinoid concentrations and compared with arachidonic acid release under identical conditions. Time course studies will also be done to help elucidate the sequence of events involved. Specific inhibitors of the various enzymes will be used where possible to supplement the data obtained by direct measurement. Eicosanoids, such as prostaglandin E2, have been shown to be important mediators of cannabinoid action in humans as well as in animal models so that a detailed understanding of how their levels are elevated is of importance. The recent discovery of an endogenous cannabinoid, anandamide, that may function as a brain neuroregulator, lends even greater significance to the aims of this proposal because of its structural nature, namely, the ethanolantide derivative of arachidonic acid. The significance of the anandamide discovery to the "prostaglandin hypothesis" of cannabinoid action is not clear at this time, however, it is unlikely that the eicosanoid nature of anandamide is only coincidental. A better understanding of THC/anandamide biochemical actions could improve our understanding of processes such as memory, motor coordination cognitive ability, time sense and self-perception. In addition, some insight might be gained into the question of the molecular basis for cannabis abuse. The recent reports on the increasing use of marihuana, especially among preadolescents, has again focussed attention on efforts to understand the actions of this drug.

本申请中提出的实验的目的是确定 导致大麻素诱导的花生四烯酸的生化事件 发布。即将开展的研究将集中于信号 大麻素与其受体结合后的转导过程。这 在此过程中受体介导的问题将由 除了更多的使用反义寡脱氧核苷酸 传统的配体结合方法。寡核苷酸的设计目的是减少， 或完全消除大麻素受体基因的表达 并与随后花生四烯酸释放的变化进行比较。期限 大麻素包括外源性物质，例如 THC、 和内源性分子，如 anandamide。用于这些的系统 研究将分离出完整的细胞，例如小鼠腹膜 巨噬细胞、神经母细胞瘤和中国仓鼠卵巢系。这 实验旨在确定磷脂酶 D、蛋白质的作用 激酶 C、MAP 激酶和磷脂酶 A2 在介导大麻素中的作用 刺激花生四烯酸的释放以及随后产生 类二十烷酸。酶活性的直接测量将在 大麻素浓度范围并与花生四烯酸进行比较 相同条件下释放。时间课程研究也将完成 帮助阐明所涉及事件的顺序。特异性抑制剂 将尽可能使用各种酶来补充数据 通过直接测量获得。 类二十烷酸，如前列腺素 E2，已被证明很重要 大麻素在人类和动物模型中的作用介质 详细了解他们的水平是如何提高的 重要性。最近发现了一种内源性大麻素 anandamide， 可能起到大脑神经调节器的作用，提供更大的帮助 由于其结构性，对该提案的目标具有重要意义 性质，即花生四烯酸的乙醇抗肽衍生物。这 阿那达酰胺的发现对“前列腺素假说”的意义 目前大麻素的作用尚不清楚，但可能性不大 anandamide 的类二十烷酸性质只是巧合。更好的 了解 THC/anandamide 生化作用可以提高我们的 对记忆、运动协调、认知等过程的理解 能力、时间观念和自我认知。此外，一些见解可能 深入探讨大麻滥用的分子基础问题。这 最近关于大麻使用量增加的报告，特别是在 青春期前的儿童，再次将注意力集中在努力理解 这种药物的作用。