Research reportEvidence for the involvement of metabotropic glutamatergic, neurokinin 1 receptor pathways and protein kinase C in the antinociceptive effect of dipyrone in mice
Introduction
Dipyrone (metamizol), a pirazolone derivative classified as a non-steroidal anti-inflammatory agent, is widely used in most countries as an antipyretic and analgesic drug. Recently, Chandrasekharan et al. [13] proposed that dipyrone could act through a newly defined mechanism via interaction with COX-3, a cyclooxygenase-1 variant that is abundant in cerebral cortex. However, despite many experimental results [1], [4], [15], [16], [19], [20], [24], [25], [31], [42], [48] taken as evidence of nociception, the precise mechanism through which dipyrone exerts these clinically relevant actions remains uncertain.
Data from our laboratory have shown that dipyrone elicits significant and dose-related spinal and supraspinal antinociception when assessed against formalin, capsaicin and glutamate-induced nociception in mice [4]. The mechanism associated with the antinociceptive action of dipyrone does not seem to be related to its interaction with opioids, GABAB or serotonin receptors, ATP-sensitive potassium channels, nitric oxide or G proteins sensitive to pertussis toxin [4]. Besides antagonising glutamate-induced pain, dipyrone partly inhibited glutamate binding sites in brain cortex [4]. Recently, Ortiz et al. [34] confirmed the absence of involvement of ATP-sensitive K+ channels in dipyrone antinociception, but reported that two blockers of the large and small-conductance-calcium-activated K+ channels, apamin and charybdotoxin, dose-dependently blocked dipyrone-induced antinociception in the formalin test. In contrast, Alves and Duarte [3] found that glibenclamide, an ATP-sensitive K+ channel blocker, but not apamin or charybdotoxin, injected into the paw, was capable of inhibiting the antinociception caused by i.pl. injection of dipyrone against prostaglandin E2-induced hyperalgesia. Thus, depending on the model used to induce nociception and on the route of administration, different mechanisms seem to be involved in the antinociceptive action of dipyrone in rodents.
Glutamatergic receptors are widespread in the central nervous system and are divided into two major types, ionotropic (coupled to membrane ion channels) and metabotropic (mGluRs, coupled to G proteins). Recent evidence obtained using glutamatergic-selective receptor agonists and antagonists indicates that all the subtypes of ionotropic (N-methyl-d-aspartate (NMDA), α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA) and kainate) and metabotropic (groups I, II and III) glutamatergic receptors play relevant roles in the establishment or maintenance of painful states, either in acute or chronic nociception [32], [40], [41], [49] (for review, see Ref. [9]). Furthermore, evidence also suggests that glutamate and some neuropeptides, especially substance P (SP), co-exist in both large and small diameter peripheral sensory fibres [14], [43], [46]. In addition, SP increases the glutamate-induced nociception [10] and the two neurotransmiters can act synergistically in the transmission of pain [5], [26], [38], [45].
In the present study, we sought to examine further the mechanisms through which dipyrone causes antinociception in mice. For this purpose we used selective glutamatergic receptor agonists, substance P and a direct activator of protein kinase C (PKC).
Section snippets
Animals
Male Swiss mice (25–35 g) were used, housed at 22±2 °C under a 12:12-h light/dark cycle and with access to food and water ad libitum. Experiments were performed during the light phase of the cycle. Animals were acclimatised to the laboratory for at least 1 h before testing and were used once during the experiments. Experiments reported in this study were carried out in accordance with current guidelines for the care of laboratory animals and ethical guidelines for investigation of experimental
Effect of dipyrone on spinal excitatory amino acids (EAAs)- or substance P (SP)-induced biting response
Fig. 1A and B shows that dipyrone (15–500 mg/kg), given i.p., 30 min prior resulted in a significant and dose-related inhibition of the biting responses to glutamate (175 nmol/site, i.t.) or trans-ACPD (2.6 nmol/site, i.t.). The inhibition observed was 41.8±15.4% and 80.8±4.3%, respectively. The calculated mean ID50 value for the antinociceptive effect of dipyrone against trans-ACPD-induced biting was 29.2 (22.8–37.4) mg/kg. Given by i.p. route, dipyrone (60–180 mg/kg) also inhibited by
Discussion
The results of the present study extend previous data from our group [4], suggesting that the glutamatergic system plays a critical role in the peripheral and spinal mechanisms through which dipyrone exerts its antinociceptive action in mice. By using selective glutamate receptor agonists we have confirmed previous data from the literature showing that intrathecal administration of trans-ACPD (2.6 nmol/i.t.), kainate (110 pmol/i.t.), AMPA (135 pmol/i.t.) and NMDA (450 pmol/i.t.) results in
Acknowledgements
This study was supported by grants from the Conselho Nacional de Desenvolvimento Cientı́fico e Tecnológico (CNPq), Financiadora de Estudos e Projetos (FINEP), and Programa de Apoio aos Núcleos de Excelência (PRONEX), Brazil. J.S. Siebel is an undergraduate Pharmacy student and she thanks CNPq for support in the form of a studentship.
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