FAAH inhibition in migraine pain
| Main Authors: | Greco Rosaria, Demartini Chiara, Tassorelli Cristina, Zanaboni Anna, Tumelero Elena, Reggiani Angelo, Misto Alessandra, Piomelli Daniele |
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| Format: | info dataset |
| Terbitan: |
, 2021
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| Subjects: | |
| Online Access: |
https://zenodo.org/record/5176109 |
Daftar Isi:
- This dataset comprises the findings obtained in the study aimed at investigating the relevance of fatty-acid amide hydrolase (FAAH) inhibition in the nitroglycerin (NTG) animal model of migraine, by testing the effects of the globally active FAAH inhibitor URB597. The attention was focused on mapping neuronal c-Fos protein expression, measurement of anandamide (AEA) and palmitoylethanolamide (PEA) levels in brain areas and in trigeminal ganglia, evaluation of pain-related trigeminal behavior and quantification of molecular mediators in rats that received URB597 (2 mg/kg i.p.) either before or after NTG administration (10 mg/kg, i.p.). Evaluation of c-Fos expression by immunohistochemistry: cell counts of individual brain nuclei were made from every sixth section throughout their rostrocaudal extent for each rat. The nuclei evaluated were identified with the rat brain atlas Paxinos and Watson 4th edition. Trigeminal nucleus caudalis (TNC): from bregma −14.08 mm (interaural −5.08 mm) until −17 mm (interaural −8 mm); nucleus tractus solitarii (NTS): from bregma −13.24 mm (interaural −4.24 mm) until −13.68 mm (interaural −4.68 mm); locus ceruleus (LC): from bregma −9.80 mm (interaural −0.80 mm) until −10.04 mm (interaural −1.04 mm); parabrachial nucleus (PAB): from bregma −8.80 mm (interaural 0.20 mm) until −9.80 mm (interaural −0.50 mm); periaqueductal gray (PAG): from bregma −7.80 mm (interaural 1.20 mm) until −8.80 mm (interaural 0.20 mm). Measurement of AEA and PEA levels: medulla, cervical spinal cord and trigeminal ganglia were homogenized in cold methanol (2 ml) containing AEA-d4 and PEA-d4 as internal standards. After extraction the lipids were measured using a Xevo TQ UPLC-MS/MS system equipped with a reversed-phase BEH C18 column (2.1 × 50 mm, 1.7 μm particle size) (Waters, Milford, USA). Pain-related behavior in the orofacial formalin test: the face rubbing was measured counting the seconds the animal spent grooming the injected area (upper lip, lateral to the nose) with the ipsilateral forepaw or hindpaw 0–6 min (Phase I) or 12–45 min (Phase II) after formalin injection (50 μl, s.c.). The observation time was divided into 15 blocks of 3 min each. mRNA expression levels: calcitonin gene-related peptide (CGRP), neuronal nitric oxide synthase (nNOS), interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-alpha) mRNA were evaluated in in medulla, cervical spinal cord and trigeminal ganglia; whereas, in dura mater were evaluated IL-1β and inducible NOS (iNOS). 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.
- Funding: Italian Ministry of Health to IRCCS Mondino Foundation, Pavia, Italy (RF2013-02355704)