Anti-inflammatory Effects of Novel Minor Cannabinoids and Terpenes on Cellular and Murine Models of HIV and HIV Proteins

摘要 大约50%的艾滋病毒感染者患有顽固性疼痛，许多人自我报告说 通过吸食大麻来缓解症状。然而，关于大麻的影响的数据仍然有限。 在感染艾滋病毒、接受购物车治疗的人群中。研究最多的化合物是Δ9-THC，它具有精神活性 以及限制治疗潜力的成瘾效应。然而，有120多种次要的大麻素和200多种萜类 在大麻中，我们著名的大麻研究实验室发现，许多大麻中含有非精神活性物质， 治疗效果。我们确定了大麻成分可以减少HIV介导的炎症/星形胶质细胞增生症 细胞培养和HIV-蛋白介导的小鼠内脏痛。我们假设几种少量的大麻类物质和 萜类化合物将补充CART，以改善由传染性艾滋病毒引起的病毒血症、炎症和细胞毒性- 1.此外，我们预计其中几种化合物可以改善炎症，机械，内脏， 以及由CART或HIV-1蛋白促进的神经病理性疼痛状态。目标1将确定重要的大麻 在体外调节艾滋病毒介导的病毒血症和炎症的成分和靶点。利用我们广泛的 大麻素、萜烯和挥发油，我们将筛选出高大麻二醇(CBD)混合物和44纯大麻二醇 微量的大麻素和萜类化合物对模拟的人外周血单个核细胞或小胶质细胞有作用。 感染或感染HIV-1IIIB或HIV-1BaL。将评估病毒血症、细胞因子产生和细胞毒性 随后将使用CB1、CB2和GPR55的拮抗剂来研究其药效学机制。 目的研究小麻素和萜类化合物的体内抗伤害性和镇痛作用。 与HIV-1蛋白/CART相关的炎症性、热性、内脏和神经性疼痛的啮齿动物模型。剂量- 大麻红素的抗炎、抗痛敏和抗伤害性依赖作用，10β-羟基- 8-四氢大麻酚和β-石竹烯将是焦点的纯化合物，与高CBD提取物一起 (以及目标1中确定的其他抗炎线索)。这些成分将在男性和女性中进行评估 表达(或不表达)HIV-1Tat或gp120蛋白的转基因小鼠。小鼠将被维持在 CART评估潜在的改善或与大麻素相关结果的相互作用。目标3将决定 大麻素影响中枢和肠道神经系统炎症的重要机制 和在HIV-1蛋白/购物车暴露后的萜烯。与疼痛相关的脑区、脊髓、背根 在AIM 2中收集的神经节、回肠、水肿和血浆将被评估细胞因子的产生。脑和回肠 将通过[35S]GTPγS试验以及神经元形态和功能来评估CB1或CB2G蛋白的活性 单核细胞来源的细胞和星形胶质细胞增生症。这些研究将提供对次要疾病的机制的洞察 大麻素和萜类化合物的作用，将揭示其抗病毒、抗炎和抗伤害性的潜力 非精神活性大麻成分，并将阐明艾滋病毒相关和非艾滋病毒相关的治疗方法， 顽固的疼痛状态。

Summary Approximately 50% of HIV-infected patients suffer from intractable pain and many individuals self-report the consumption of cannabis for alleviating their symptoms. However, data are still limited on the effects of cannabis in the HIV-infected, cART-treated population. The compound most studied is Δ9-THC, which exerts psychoactive and addictive effects that limit therapeutic potential. Yet, there are over 120 minor cannabinoids and 200 terpenes in cannabis and our renowned Marijuana Research Laboratory has found many of them hold non-psychoactive, therapeutic effects. We identified cannabis constituents that reduce HIV-mediated inflammation/astrogliosis in cell culture and HIV-protein mediated visceral pain in mice. We hypothesize that several minor cannabinoids and terpenes will complement cART to ameliorate viremia, inflammation, and cytotoxicity caused by infectious HIV- 1. Moreover, we anticipate several of these compounds to ameliorate the inflammatory, mechanical, visceral, and neuropathic pain states promoted by cART or HIV-1 proteins. Aim 1 will determine the important cannabis constituents and targets that modulate HIV-mediated viremia and inflammation in vitro. Utilizing our extensive library of cannabinoids, terpenes, and volatile oils, we will screen a high-cannabidiol (CBD) mixture and 44 pure minor cannabinoids and terpenes against human peripheral blood mononuclear cells or microglia that are mock- infected or infected with HIV-1IIIB or HIV-1BaL. Viremia, cytokine production, and cytotoxicity will be assessed and the pharmacodynamic mechanisms will be subsequently examined using antagonists to CB1, CB2, and GPR55. Aim 2 will determine the in vivo anti-nociceptive and analgesic effects of minor cannabinoids and terpenes in rodent models of HIV-1 protein/cART-related inflammatory, thermal, visceral, and neuropathic pain. Dose- dependent anti-inflammatory, anti-hyperalgesic, and anti-nociceptive effects of cannabichromene, 10β-hydroxy- 8-tetrahydrocannabinol, and β–caryophyllene will be the pure compounds of focus, along with High-CBD extract (and other anti-inflammatory leads identified in Aim 1). These constituents will be assessed in male and female transgenic mice that express (or do not express) the HIV-1 Tat or gp120 proteins. Mice will be maintained on cART to assess potential improvements or interactions with cannabinoid-related outcomes. Aim 3 will determine the important central and enteric nervous system inflammatory mechanisms that are influenced by cannabinoids and terpenes following HIV-1 protein/cART exposure. Pain-related brain regions, spinal cord, dorsal root ganglion, ileum, edema, and plasma collected in Aim 2 will be assessed for cytokine production. Brain and ileum will be assessed for CB1 or CB2 G-protein activity via [35S]GTPγS assay as well as neuron morphology and monocyte-derived cell and astrogliosis. TThese studies will provide insight into the mechanisms by which minor cannabinoids and terpenes act, will reveal the anti-viremic, anti-inflammatory, and antinociceptive potential of non-psychoactive cannabis constituents, and will elucidate therapeutics for HIV-related and non-HIV-related, intractable pain states.