Cannabinoid receptor-mediated antinociception with acetaminophen drug combinations in rats with neuropathic spinal cord injury pain

Abstract

Pre-clinical evidence demonstrates that neuropathic spinal cord injury (SCI) pain is maintained by a number of neurobiological mechanisms, suggesting that treatments directed at several pain-related targets may be more advantageous compared to a treatment focused on a single target. The current study evaluated the efficacy of the non-opiate analgesic acetaminophen, which has several putative analgesic mechanisms, combined with analgesic drugs used to treat neuropathic pain in a rat model of below-level neuropathic SCI pain. Following an acute compression of the mid-thoracic spinal cord, rats exhibited robust hind paw hypersensitivity to innocuous mechanical stimulation. Fifty percent antinociceptive doses of gabapentin, morphine, tramadol or memantine were combined with an ineffective dose of acetaminophen; acetaminophen alone was not antinociceptive. The combination of acetaminophen with either tramadol or memantine resulted in an additive antinociceptive effect. Acetaminophen combined with either morphine or gabapentin, however, resulted in supra-additive (synergistic) efficacy. One of the analgesic mechanisms of acetaminophen is inhibiting the uptake of endocannabinoids from the extracellular space. Pre-treatment with AM251, a cannabinoid-1 receptor (CB1) antagonist, significantly diminished the antinociceptive effect of the acetaminophen + gabapentin combination. Pre-treatment with AM630, a cannabinoid-2 receptor (CB2) antagonist, did not have an effect on this combination. By contrast, both AM251 and AM630 reduced the efficacy of the acetaminophen + morphine combination. None of the active drugs alone were affected by either CB receptor antagonist. The results imply that modulation of the endocannabinoid system in addition to other mechanisms mediate the synergistic antinociceptive effects of acetaminophen combinations. Despite the presence of a cannabinoid mechanism, synergism was not present in all acetaminophen combinations. The combination of currently available drugs may be an appropriate option in ameliorating neuropathic SCI pain if single drug therapy is ineffective.

Figure 1

Comparison of the potencies of gabapentin (A), memantine (B), morphine (C) and tramadol (D) alone and in combination with acetaminophen (APAP). The horizontal axis is the dose of the active drug (in mg/kg) and the vertical axis is percent maximum possible effect (%). For comparison, the effects of 100 mg/kg APAP alone (∇) at 90 min (A) and 60 min (B) post-injection are shown. Data are expressed as mean ± S.E.M. n = 6-8/group.

Figure 2

Effect of CB receptor antagonists on the antinociceptive effect of acetaminophen+gabapentin. Rats were pretreated with either vehicle (veh) or antagonist 30 min prior to a second injection of either vehicle or APAP+gabapentin combination. Rats were tested 90 min post-injection of either acetaminophen+gabapentin (APAP+GP) or vehicle. (A) A robust antinociception was observed in APAP+GP rats pretreated with vehicle. However, pretreatment with AM251 significantly reduced the antinociceptive effect of APAP+GP. (B) Pretreatment with AM630 did not affect APAP+GP antinociception. Data are expressed as mean ± S.E.M. *p < 0.05 vs. veh/veh, #p < 0.05 vs. veh/APAP+GP; n = 7-8/group.

Figure 3

Effect of CB receptor antagonists on the antinociceptive effect of acetaminophen+morphine. Rats were pretreated with either vehicle (veh) or antagonist 30 min prior to a second injection of either vehicle or APAP+morphine combination. Rats were tested 60 min post-injection of either acetaminophen+morphine (APAP+Mor) or vehicle. (A) A robust antinociception was observed in APAP+Mor rats pretreated with vehicle. Pretreatment with AM251 significantly reduced the antinociceptive effect of APAP+Mor, however, withdrawal thresholds were still significantly elevated compared to the AM251/veh group. (B) Pretreatment with AM630 significantly reduced the antinociceptive effect of APAP+Mor, however, withdrawal thresholds were still significantly elevated compared to the AM630/veh group. Data are expressed as mean ± S.E.M. *p < 0.05 vs. veh/veh; #p < 0.05 vs. veh/APAP+Mor; ˆp < 0.05 vs. antagonist/veh; n = 7/group.

Figure 4

Effect of CB receptor antagonists on the antinociceptive effect of acetaminophen+tramadol. Rats were pretreated with either vehicle (veh) or antagonist 30 min prior to a second injection of either vehicle or acetaminophen+tramadol combination. Rats were tested 90 min post-injection of either acetaminophen+tramadol (APAP+Tram) or vehicle. (A) A robust antinociception was observed in APAP+Tram rats pretreated with vehicle. However, pretreatment with AM251 significantly reduced the antinociceptive effect of APAP+Tram. (B) Pretreatment with AM630 did not significantly change the antinociceptive effect of APAP+Tram. Data are expressed as mean ± S.E.M. *p < 0.05 vs. veh/veh; #p < 0.05 vs. veh/APAP+Tram; n = 7/group.

Figure 5

Effect of a μ-opioid receptor antagonist on the antinociceptive effect of acetaminophen+tramadol. Rats were pretreated with either vehicle (veh) or antagonist 30 min prior to a second injection of either vehicle or acetaminophen+tramadol combination. Rats were tested 90 min following the second injection of either APAP+Tram or vehicle. Pretreatment with naloxone significantly reduced the antinociceptive effect of APAP+Tram. Data are expressed as mean ± S.E.M. *p < 0.05 vs. veh/veh; #p < 0.05 vs. veh/APAP+Tram; n = 7/group.