Cannabidiol and terpenoid interactions in amygdalar regulation of pain states

项目总结 慢性疼痛影响多达三分之一的美国人口，依赖阿片类药物治疗慢性疼痛 疼痛在很大程度上导致了阿片类药物的流行。开发替代的止痛疗法对 减少阿片类药物的使用，而植物大麻二醇(CBD)是一个很有前途的候选药物。我们有 结果表明，CBD有能力在3周内减轻小鼠的慢性神经病理性疼痛样反应。 萜类化合物(如β-石竹烯)，也在大麻植物中发现，是一种潜在的第二类 大麻中的药理活性化合物可能有止痛作用，尽管它们 体外和体内的药效学特性还知之甚少。同样，还有一个不完整的 了解CBD和萜类化合物是如何单独或联合产生抗伤害性的(“环境”)， 而它们的脊椎上“疼痛控制”的神经药理机制仍不清楚。建议数 实验将首先测试CBD和萜类化合物的特定组合是否可以产生短期的 没有耐受性的炎症性和长期神经病理性的抗伤害感受。然后，我们将建立 CBD和萜类化合物在疼痛的关键脑核-杏仁基底外侧核中的作用。最后，我们会 测定CBD和萜类化合物在体外和体内的药理和生化信号谱 活着。在目标1中，我们将在我们的明胶中使用CBD和萜类化合物的混合物，调查员管理以及我们的明胶 自我给药模型，并测量部分坐骨神经结扎后的消耗和疼痛评分 确定CBD和/或萜类化合物是否对长时间的疼痛有止痛作用。我们还将核实 这些混合物本身并不具有回报，这对药物滥用责任至关重要。在目标2中，我们将 用单细胞测试杏仁核是否是CBD/萜类止痛作用的关键脑回路部位 疼痛状态下杏仁核神经元的钙成像，同时局部注射CBD和萜类化合物 进入杏仁核产生止痛作用。此外，我们将敲除大麻素受体和其他假定的 CBD/萜类化合物在BLA内的作用部位，以确定每个部位在产生行为和 生理反应。在目标3中，我们将使用体外和体内系统来确定重要的生化 CBD的特征和萜类化合物对其可能的受体靶点的作用。我们将测量细胞信号活动 通过动态质量重分配(DMR)、MAP激酶信号转导和活性氧物种的产生。 这些目标将使公众健康了解长期使用大麻二醇/萜类化合物的好处和风险，如 并提供关键的机械洞察，将帮助开发和理解定制的药物 可以利用大麻治疗慢性疼痛的方法来获得治疗效益。

PROJECT SUMMARY Chronic pain affects up to one third of the United States population, and the reliance on opioids to treat chronic pain has contributed substantially to the opioid epidemic. Developing alternative pain therapies is critical to reducing the use of opioids, and the phyto-cannabinoid cannabidiol (CBD) is a promising candidate. We have shown that CBD has the ability to reduce chronic neuropathic pain-like responses in mice over 3 weeks. Terpenoids (eg beta-caryophyllene), also found in the cannabis plant, are a potential second class of pharmacologically active compounds in cannabis with possible analgesic benefits, although their pharmacodynamic properties in vitro and in vivo are poorly understood. Likewise, there is an incomplete understanding of how CBD and terpenoids produce anti-nociception, by themselves or combined (“entourage”), and their supraspinal neuropharmacological mechanisms of “pain control” remain unknown. The proposed experiments will first test whether specific combinations of CBD and terpenoids can produce short-term inflammatory and long-term neuropathic antinociception without tolerance. We will then establish the action of CBD and terpenoids in a critical brain nucleus for pain, the basolateral amygdala. Finally, we will determine the pharmacological and biochemical signaling profiles of CBD and terpenoids in vitro and in vivo. In Aim 1, we will use mixtures of CBD and terpenoids, investigator administered as well as in our gelatin self-administration model, and measure both consumption and pain scores after partial sciatic nerve ligation to determine if CBD and/or terpenoids provide analgesic benefit over protracted periods of pain. We will also verify that these mixtures are not inherently rewarding, which is critical for substance abuse liability. In Aim 2, we will test whether the amygdala is a critical brain circuit site for CBD/terpenoid analgesic action using single cell calcium imaging of amygdalar neurons during pain states, in parallel with local injections of CBD and terpenoids into the amygdala to produce analgesia. Further, we will knock out cannabinoid receptors and other putative sites of CBD/terpenoid action within the BLA to establish necessity of each in producing behavioral and physiological responses. In Aim 3, we will use in vitro and in vivo systems to determine the important biochemical features of CBD and terpenoid action at their putative receptor targets. We will measure cellular signaling activity via dynamic mass redistribution (DMR), MAP Kinase signaling, and the generation of reactive oxygen species. These aims will inform public health about the benefits and risks of long-term cannabidiol/terpenoid usage, as well as providing crucial mechanistic insight that will help develop and understand whether tailored medicinal cannabis approaches for chronic pain can be harnessed for therapeutic benefit.