Nitrous oxide suppresses tonic and phasic nociceptive behaviors but not formalin-induced c-Fos expression in the rat spinal cord dorsal horn.

Abstract

The aim of this study was to investigate the anesthetic and analgesic effects of subanesthetic concentration of nitrous oxide and to compare these effects with halothane and fentanyl. The antinociceptive effects were assessed in male Sprague-Dawley rats by behavioral responses to phasic and tonic nociceptive stimulations and biochemical index of pain, formalin-induced Fos-like immunoreactivity (Fos-LI), in spinal cord dorsal horn. Neurological functions (proprioception, mental status and motor function) were monitored to determine whether or not behavioral responses were impaired by anesthetic action of the treatment. Four groups of rats treated with: (1) saline, (2) 75% nitrous oxide (0.5 MAC), (3) 0.5% halothane (0.5 MAC) and (4) fentanyl 100 micrograms/kg were subject either to graded intensity of CO2 laser stimulation (phasic pain) or s.c. injection of 50 microliters 2.5% formalin (tonic pain) in two separate studies. All rats in the tonic pain study were killed for immunohistochemistry at 60 min after formalin injection. Maximal counts of Fos-LI labelled neurons in rat spinal cord dorsal horn were compared according to the laminar distribution. We found that all rats exhibited normal righting reflexes regardless of whatever treatment. Nitrous oxide and halothane greatly impaired mental status and motor function, indicating that both agents could induce a modest degree of sedation and paresis at subanesthetic concentrations. Fentanyl increased the threshold level to noxious thermal stimulation, and reduced the formalin-induced licking/biting behaviors and the number of Fos-LI labelled neurons which are predominantly found in the neck of the dorsal horn. Nitrous oxide and halothane increased the thermal nociceptive threshold, suppressed licking/biting behavior in both early and late phases of the formalin test. Unlike fentanyl, nitrous oxide and halothane failed to suppress c-fos expression. The extent and pattern of nitrous oxide-induced antinociception was identical to halothane, which is known to have little or no analgesic effect. The lack of attenuated biochemical response to tonic pain stimulation may suggest that nitrous oxide fails to suppress spinal sensitization despite its potent inhibition on behavioral hyperalgesia. These findings suggest that, at the spinal level, subanesthetic concentration of nitrous oxide greatly attenuates nociceptive behaviors mainly due to a non-selective suppression of behavioral responses that are commonly associated with inhalation anesthetic drugs.