Identification of cytokines and signaling proteins differentially regulated by sumatriptan/naproxen

Abstract

Objectives: The goal of this study was to use protein array analysis to investigate temporal regulation of stimulated cytokine expression in trigeminal ganglia and the spinal trigeminal nucleus in response to co-treatment of sumatriptan and naproxen sodium or individual drug.

Background: Activation of neurons and glia in trigeminal ganglia and the spinal trigeminal nucleus leads to increased levels of cytokines that promote peripheral and central sensitization, which are key events in migraine pathology. While recent clinical studies have provided evidence that a combination of sumatriptan and naproxen sodium is more efficacious in treating migraine than either drug alone, it is not well understood why the combination therapy is superior to monotherapy.

Methods: Male Sprague-Dawley rats were left untreated (control), injected with capsaicin, or pretreated with sumatriptan/naproxen, sumatriptan, or naproxen for 1 hour prior to capsaicin. Trigeminal ganglia and the spinal trigeminal nucleus were isolated 2 and 24 hours after capsaicin or drug treatment, and levels of 90 proteins were determined using a RayBio® Label-Based Rat Antibody Array (RayBiotech, Norcross, GA, USA).

Results: Capsaicin stimulated a >3-fold increase in expression of the majority of cytokines in trigeminal ganglia at 2 hours that was sustained at 24 hours. Significantly, treatment with sumatriptan/naproxen almost completely abolished the stimulatory effects of capsaicin at 2 and 24 hours. Capsaicin stimulated >3-fold expression of more proteins in the spinal trigeminal nucleus at 24 hours when compared to 2 hours. Similarly, sumatriptan/naproxen abolished capsaicin stimulation of proteins in the spinal trigeminal nucleus at 2 hours and greatly suppressed protein expression 24 hours post-capsaicin injection. Interestingly, treatment with sumatriptan alone suppressed expression of different cytokines in trigeminal ganglia and the spinal trigeminal nucleus than repressed by naproxen sodium.

Conclusion: We found that the combination of sumatriptan/naproxen was effective in blocking capsaicin stimulation of pro-inflammatory proteins implicated in the development of peripheral and central sensitization in response to capsaicin activation of trigeminal neurons. Based on our findings that sumatriptan and naproxen regulate expression of different proteins in trigeminal ganglia and the spinal trigeminal nucleus, we propose that these drugs function on therapeutically distinct cellular targets to suppress inflammation and pain associated with migraine.

Figure 1

Sumatriptan/naproxen combination treatment suppresses capsaicin stimulated changes in the number and level of cytokine expression in trigeminal ganglia tissues at 2 (A) and 24 hours (B). Trigeminal ganglia were obtained from animals treated with capsaicin (CAP) alone, or cotreated with capsaicin and naproxen (N), sumatriptan (S), or the combination of both drugs (S/N). The height of each box corresponds to the number of proteins significantly (p < 0.05) elevated above untreated control levels, whose mean was made equal to one. The average fold change above control levels is illustrated by different colors. n = 3 independent experiments for each condition.

Figure 2

Sumatriptan/naproxen combination treatment suppresses capsaicin stimulated changes in the number and level of cytokine expression in spinal trigeminal nuclei at 2 (A) and 24 hours (B). Sections of the upper spinal cord containing the spinomedulary junction were obtained from animals treated with capsaicin (CAP) alone, or cotreated with capsaicin and naproxen (N), sumatriptan (S), or the combination of both drugs (S/N). The height of each box corresponds to the number of proteins significantly (p < 0.05) elevated above untreated control levels, whose mean was made equal to one. The average fold change above control levels is illustrated by different colors. n = 3 independent experiments for each condition.