Characterization of the biochemical and behavioral effects of cannabidiol: implications for migraine

This dataset comprises the findings obtained in the study aimed at investigating the anti-migraine effects of cannabidiol (CBD) in the migraine rat models based by nitroglycerin (NTG) administration. We assayed CBD distribution (15 and 30mg/kg, i.p.) in plasma and in cranial areas related to migraine pain in male Sprague Dawley rats treated chronically (5 days). Successively, we tested CBD activity on the behavioral and biochemical effects induced in the acute and the chronic migraine animal models by NTG administration. In the acute migraine model, rats received CBD (15 mg or 30 mg/kg, i.p) 3 h after NTG (10 mg/kg i.p.) or vehicle injection. In the chronic migraine model, rats were treated with CBD and NTG every other day over nine days with the following doses: CBD 30 mg/kg i.p., NTG 10 mg/kg i.p. We evaluated behavioral parameters with the open field and the orofacial formalin tests. We explored the fatty acid amide hydrolase (FAAH) gene expression, cytokines mRNA and protein levels in selected brain areas and CGRP serum level. CBD levels in the meninges, trigeminal ganglia, cervical spinal cord, medulla pons, and plasma were higher 1 h after the last treatment than after 24 h, suggesting that CBD penetrates but does not accumulate in these tissues. In the acute model, CBD significantly reduced NTG-induced trigeminal hyperalgesia and CGRP and cytokine mRNA levels in peripheral and central sites. In the chronic model, CBD caused only a significant decrease in NTG-induced IL-6 protein levels in the medulla-pons, and trigeminal ganglion. It also reduced CGRP serum levels. In both experimental conditions, there was no modulation of anxiety, motor/exploratory behavior, or grooming. The in vivo and ex vivo assessments were: 1) mRNA expression levels: CGRP, SP, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and FAAH in cervical spinal cord, medulla-pons and trigeminal ganglia. mRNA levels were measured by rt-PCR. All samples were assayed in triplicate and gene expression levels were calculated according to 2−∆∆Ct = 2− (∆Ct gene − ∆Ct housekeeping gene) formula by using Ct (cycle threshold) values. 2) Central and peripheral CBD levels were quantified in rat brain tissue and plasma samples using a previously proven online solid phase extraction (SPE) high-performance liquid chromatography (HPLC) method coupled with tandem mass spectrometry (MS/MS). 3) Cytokines and CGRP protein levels were measured using the ELISA kits. The samples' measured absorbance was compared to a standard curve using a microplate reader. 4) Behavior Formalin test= the face rubbing was measured counting the seconds the animal spent grooming the injected area (seconds). 5) open field test= distance (expressed in meters) travelled in the apparatus; time spent (expressed in seconds) in the center of the apparatus; number of rearing; time spent in grooming behavior (expressed in seconds). Results in brief These data show that CBD modulates migraine-related nociceptive transmission, likely via a complex signaling mechanism involving different pathways.本数据集源自一项旨在探究大麻二酚（cannabidiol, CBD）对基于硝酸甘油（nitroglycerin, NTG）给药构建的偏头痛大鼠模型抗偏头痛效应的研究，收录了该研究的全部实验结果。我们对连续5天慢性给药的雄性Sprague Dawley大鼠，检测了其血浆及偏头痛疼痛相关颅区的CBD分布情况，给药剂量为15与30mg/kg，腹腔注射（intraperitoneal, i.p.）。随后，我们评估了CBD对急性及慢性偏头痛动物模型中NTG给药诱导的行为学与生化效应的干预活性。在急性偏头痛模型中，大鼠于NTG（10mg/kg腹腔注射）或赋形剂给药3小时后，接受CBD（15或30mg/kg腹腔注射）处理。在慢性偏头痛模型中，大鼠每间隔一日给药，持续9天，给药方案如下：CBD 30mg/kg腹腔注射，NTG 10mg/kg腹腔注射。我们采用旷场实验（open field test）与口面部福尔马林实验，对行为学参数进行评估。我们检测了选定脑区的脂肪酸酰胺水解酶（fatty acid amide hydrolase, FAAH）基因表达、细胞因子mRNA及蛋白水平，以及血清降钙素基因相关肽（calcitonin gene-related peptide, CGRP）水平。末次给药后1小时，大鼠脑膜、三叉神经节、颈脊髓、延髓脑桥以及血浆中的CBD水平均高于给药后24小时，提示CBD可穿透上述组织但未在其中蓄积。在急性模型中，CBD可显著降低NTG诱导的三叉神经痛觉过敏，以及外周与中枢位点的CGRP与细胞因子mRNA水平。在慢性模型中，CBD仅可显著降低延髓-脑桥与三叉神经节中NTG诱导的白细胞介素-6（interleukin-6, IL-6）蛋白水平，同时可降低血清CGRP水平。在两种实验条件下，均未观察到焦虑、运动/探索行为或理毛行为的显著变化。体内与离体实验评估内容如下：1. mRNA表达水平：检测颈脊髓、延髓-脑桥及三叉神经节中的CGRP、P物质（substance P, SP）、IL-6、肿瘤坏死因子-α（tumor necrosis factor-α, TNF-α）及FAAH的mRNA水平。采用实时荧光定量PCR（rt-PCR）检测mRNA水平，所有样本均设三次重复，基因表达水平按照2−∆∆Ct = 2−(∆Ct靶基因 − ∆Ct持家基因)公式，以循环阈值（cycle threshold, Ct）值进行计算。2. 采用此前已验证的在线固相萃取（solid phase extraction, SPE）-高效液相色谱（high-performance liquid chromatography, HPLC）联用串联质谱（tandem mass spectrometry, MS/MS）法，对大鼠脑组织及血浆样本中的中枢与外周CBD水平进行定量。3. 采用酶联免疫吸附测定（enzyme linked immunosorbent assay, ELISA）试剂盒检测细胞因子与CGRP蛋白水平，通过酶标仪读取样本吸光度，并与标准曲线进行比对。4. 福尔马林实验：通过记录动物舔舐/理毛注射区域的时长（秒），量化面部擦挠行为。5. 旷场实验：记录动物在装置内的移动距离（以米计）、在装置中心区域停留的时长（以秒计）、直立次数，以及理毛行为持续时长（以秒计）。简要结果：本研究数据表明，CBD可调控偏头痛相关的伤害性感受传递，其潜在机制可能涉及多条复杂的信号通路。