NO/cGMP production is important for the endogenous peripheral control of hyperalgesia during inflammation.

Various studies have demonstrated the role of the nitric oxide (NO)/cGMP pathway in pain processing. Our group has also shown that this system participates in opioid-induced antinociception during peripheral inflammation. We have previously observed that inflammation mobilizes an endogenous opioidergic system to control hyperalgesia. Here, we investigated whether the NO/cGMP pathway underlies peripheral endogenous nociception control during inflammation. In this study, a pharmacological approach was used in conjunction with the rat paw pressure test to assess the effects of intraplantar NO synthase inhibitor NG-Nitro-l-arginine (NOArg), guanylyl cyclase inhibitor methylene blue (MB), phosphodiesterase-5 inhibitor zaprinast (ZP), or NO precursor l-arginine injection on carrageenan-induced hyperalgesia, which mimics an inflammatory process, or by prostaglandin E(2) (PGE(2)), which directly sensitizes nociceptors. Intraplantar carrageenan (62.5, 125, 250 or 500µg) or PGE(2) (0.1, 0.5 or 2µg) administration produced hyperalgesia, which manifested as a reduction in the rat nociceptive threshold to mechanical stimuli. NOArg (25, 50 or 100µg/paw) and MB (125, 250 or 500µg/paw) induced significant and dose-dependent reductions in the nociceptive threshold of carrageenan-induced (125µg/paw) hyperalgesia, but not PGE(2)-induced (0.5µg/paw) hyperalgesia. This was a local effect because it did not produce any modifications in the contralateral paw. Both Zaprinast (100, 200 or 400µg/paw) and l-arginine (100, 200 or 400µg/paw) significantly counteracted carrageenan-induced hyperalgesia (250µg/paw), yielding an increase in the nociceptive threshold compared with the control. Zaprinast (200µg/paw) or l-arginine (400µg/paw) did not produce an antinociceptive effect in the contralateral paw, indicating local action. In addition, at the same dose that was able to modify carrageenan-induced hyperalgesia, neither zaprinast nor l-arginine modified PGE(2) (2µg) injection-induced hyperalgesia of the rat paw. Taken together, these results indicate that the l-arginine/NO/cGMP pathway functions as an endogenous modulator of peripheral inflammatory hyperalgesia.

Inflammation mobilizes local resources to control hyperalgesia: the role of endogenous opioid peptides.

The aim of the present study was to investigate the mechanisms underlying the endogenous control of nociception at a peripheral level during inflammation. Using a pharmacological approach and the rat paw pressure test, we assessed the effect of an intraplantar injection of naloxone, an opioid receptor antagonist, and bestatin, an aminopeptidase inhibitor, on hyperalgesia induced by carrageenan, which mimics an inflammatory process, or prostaglandin E(2) (PGE(2)), which directly sensitizes nociceptors. Naloxone induced a significant and dose-dependent (25, 50 or 100 µg) increase in carrageenan-induced hyperalgesia, but not PGE(2)-induced hyperalgesia. Bestatin (400 µg/paw) significantly counteracted carrageenan-induced hyperalgesia, inducing an increase in the nociceptive threshold compared to control, but it did not modify hyperalgesia induced by PGE(2) injection into the rat paw. Positive ß-endorphin immunoreactivity was increased in paw inflammation induced by carrageenan in comparison with the control group. However, PGE(2) did not significantly alter the immunostained area. These results provide evidence for activation of the endogenous opioidergic system during inflammation and indicate that this system regulates hyperalgesia through a negative feedback mechanism, modulating it at a peripheral level.

Prevention by celecoxib of secondary hyperalgesia induced by formalin in rats.

Administration of formalin in rat paws results in stimulation of nociceptive pathways, which leads to an increase in the excitability of neurons present in dorsal horn. This increased neuron excitability, described as central sensitization, may result in development of inflammatory pain at a distant site of injury application, known as secondary hyperalgesia. The aim of the present study was to verify whether formalin injection in rat paws would lead to secondary hyperalgesia development, as measured by the tail-flick test. We also aimed to investigate whether celecoxib, a specific cyclooxygenase 2 (COX-2) inhibitor, would affect secondary hyperalgesia. Formalin injected into the rat paws significantly reduced the latency for a flick response in the rat tail, which characterized development of secondary hyperalgesia. In addition, formalin-induced secondary hyperalgesia was locally prevented by pre-but not post-celecoxib treatment. However, celecoxib administered spinally inhibited formalin-induced secondary hyperalgesia, either administered previously or following formalin. In contrast, piroxicam, an unspecific COX inhibitor which displays an increased selectivity towards COX-1, only prevented secondary hyperalgesia to formalin at a high dose following spinal administration. Taken together, these results suggest that COX-2 plays an important role both in the central and in the peripheral nerve sensitization following formalin administration in rat paws. They also suggested that once central sensitization starts it can no longer be blocked by a specific COX-2 inhibitor administered locally. Notwithstanding, spinal administration of a specific COX-2 inhibitor still blocks ongoing sensitization and prevents maintenance of central sensitization.

Evidence for participation of GABA(A) receptors in a rat model of secondary hyperalgesia.

We investigated the involvement of endogenous gamma-aminobutyric acid (GABA) in the modulation of secondary hyperalgesia induced by intraplantar (i.pl.) injection of 5% formalin in the rat tail-flick test. Intraplantar injection of gabamimetic drugs such as gabapentin (150-600 microg/site) or phenobarbital (20-80 microg/site) reversed secondary hyperalgesia, as measured by an increase in the tail-flick latency, thus displaying a peripheral antihyperalgesic effect. Central inhibition of the secondary hyperalgesia response by gabapentin was obtained following injection of either 200 microg intrathecally (i.t.) or 50 mg intraperitoneally (i.p.). The effects induced by gabamimetics were blocked locally or centrally by prior treatment with the specific GABA(A) receptor antagonist, bicuculline (80 ng/paw or 20 ng, i.t.). These data indicate the participation of endogenous GABA in the modulation of secondary hyperalgesia, through either a peripheral and/or a central action. They also indicate that GABA(A) receptors might be involved since a specific antagonist of these receptors (bicuculline) blocked this response.