PERIPHERAL MECHANISMS OF CANNABINOID ANTINOCICEPTION

大麻素镇痛的外围机制

• 批准号: 6453547
• 负责人: Theodore J. Price
• 金额: $ 2.14万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-10-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6453547
• 关键词: G protein; afferent nerve; analgesics; calcitonin gene related peptide; calcium flux; cannabinoid receptor; cannabinoids; capsaicin; ganglions; immunocytochemistry; immunoprecipitation; in situ hybridization; laboratory rat; neuropharmacology; neutralizing antibody; trigeminal nerve; western blottings

Cannabinoids have a long history of medicinal uses and their analgesic properties are well documented, however, their mechanisms of action, especially in the periphery, are poorly understood. In light of these analgesic effects, coupled with the untoward side effects of many currently used analgesics including the NSAIDS and opioids, we believe that careful scrutiny of the action of cannabinoid compounds at peripheral nociceptive sites is warranted. The present proposal will test the hypothesis that cannabinoids inhibit calcitonin gene related peptide (CGRP) release through cannabinoid type one receptor (CBI) mediated decreases in calcium influx. The rational for this hypothesis is that cannabinoid activation of CB1 would stimulate the dissociation of beta-gamma subunits from GalphaO which interact directly with P/Q and N type calcium channels to inhibit calcium influx. This, in turn, would decrease the available voltage sensitive calcium pool causing a reduction in the release of secretary granules from primary sensory neurons under stimulatory states. To address this hypothesis, the specific aims of this research plan will be; 1) to co-localize CB1 receptors with CGRP in small and medium diameter trigeminal ganglion primary sensory neurons through combined in situ hybridization and immunohistochemistry, 2) to evaluate the G-protein coupling properties of the CB1 receptor utilizing selective immunoprecpitation with radiolabeled CB1 ligands and commercially available G-protein alpha subunit antibodies; and 3) to examine in cultured trigeminal ganglion the second messenger pathways through which the CB1 receptor modulates capsaicin-evoked CGRP release utilizing pharmacological methodologies and calcium imaging approaches.

大麻素具有悠久的药用史，其镇痛性特性有充分的文献证明，但是，它们的作用机理，尤其是在外围的作用机理，对此知之甚少。 鉴于这些镇痛作用，再加上许多当前使用的镇痛药的不良副作用，包括NSAIDS和阿片类药物，我们认为必须仔细审查大麻素化合物在外围伤害性部位的作用。本提案将检验以下假设：大麻素抑制降钙素基因相关的肽（CGRP）通过大麻素型受体（CBI）介导的钙涌入钙的降低。 该假设的理性是，CB1的大麻素激活将刺激β-gamma亚基与Galphao的解离，而Galphao直接与P/Q和N型钙通道直接相互作用以抑制钙的涌入。 反过来，这将减少可用的电压敏感钙池，从而导致刺激状态下原代感觉神经元的秘书颗粒的释放减少。 为了解决这一假设，该研究计划的具体目的是； 1) to co-localize CB1 receptors with CGRP in small and medium diameter trigeminal ganglion primary sensory neurons through combined in situ hybridization and immunohistochemistry, 2) to evaluate the G-protein coupling properties of the CB1 receptor utilizing selective immunoprecpitation with radiolabeled CB1 ligands and commercially available G-protein alpha subunit抗体； 3）要在培养的三叉神经节中检查第二个使者途径，CB1受体通过该通路调节辣椒素诱发的CGRP释放，利用药理方法论和钙成像方法。