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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.