A7 CANNABINOID 1 AND 2 RECEPTOR AGONISTS AND MU-OPIOID RECEPTOR AGONISTS SYNERGISTICALLY INHIBIT COLONIC NOCICEPTION DURING ACUTE COLITIS

TL;DR

Combining low doses of CB2R and MOR agonists can reduce colon pain in mice with acute colitis, offering a potential new treatment for inflammatory bowel disease.

Contribution

The study reveals that CB2R agonists alone or in combination with MOR agonists inhibit colonic pain during acute colitis, a novel finding for analgesic strategies.

Findings

CB2R agonist HU-308 inhibits colonic mechanosensitivity in colitis but not in healthy mice.

Combining sub-analgesic doses of CB1R and MOR agonists reduces pain in both healthy and colitic mice.

Combining sub-analgesic CB2R and MOR agonists reduces pain only in mice with acute colitis.

Abstract

Abdominal pain is a debilitating symptom in patients with inflammatory bowel disease. Previously we have shown that combining sub-analgesic doses of cannabinoid 1 receptor (CB1R), but not cannabinoid 2 receptor (CB2R), and mu-opioid receptor (MOR) agonists synergistically inhibits colonic nociception in healthy mice. However, it is unknown whether this combination has analgesic efficacy in a pre-clinical model of colitis.

To determine the effects of combining sub-analgesic doses of CBR and MOR agonists on colonic nociception during acute colitis.

Colitis was induced in male and female C57BL/6 mice with 2.5% dextran sulfate sodium in drinking water. Extracellular afferent nerve recordings were obtained from ex vivo flat sheet preparations of mouse distal colon. Mechanosensitivity of single afferent axons was assessed via probing of the colon with a 1g von Frey hair before and after superfusion of agonists of CB1R or CB2R plus MOR. To examine effects in vivo, visceromotor response (VMR) to colorectal distention (volume range 20-80 µL) was measured via electromyography. Mice were injected intraperitoneally with vehicle, or a combination of CB1R or CB2R agonist plus morphine 30-minutes prior to VMR experiment. Data were analyzed using a one or two-way ANOVA with Bonferroni test. N denotes number of mice; n denotes number of single afferent axons.

In afferent nerve recordings, in contrast to healthy mice, the CB2R agonist HU-308 (1 µM and 3 µM) inhibited colonic mechanosensitivity in mice with colitis (1 µM: p<0.01, N=5, n=12; 3 µM: p<0.01, N=7, n=10); a lower concentration (300 nM) had no effect (p=0.52, N=6, n=10). The CB1R agonist ACEA (10 µM) reduced mechanosensitivity during acute colitis (p<0.05, n=11, N=6), whereas 100 nM (p>0.99, n=7, N=5) and 1 µM (p=0.25, n=10, N=6) had no effect. A combination of sub-analgesic concentrations of ACEA (100 nM) and DAMGO (MOR agonist, 1 nM) inhibited colonic mechanosensitivity in healthy mice (p<0.01, N=4, n=8) and during acute colitis (p<0.05, N=6, n=8). While a combination of sub-analgesic concentrations of HU-308 (300 nM) and DAMGO (1 nM) had no effect in healthy mice (p=0.70, N=4, n=8), it inhibited colonic mechanosensitivity during acute colitis (p<0.01, N=8, n=15). In VMR experiments, a combination of a sub-analgesic dose of ACEA (0.3 mg/kg) with morphine (0.3 mg/kg) reduced VMR (p<0.01, N=7) during acute colitis. Similarly, a combination of a sub-analgesic dose of HU-308 (1 mg/kg) and morphine (0.3 mg/kg) reduced VMR (p<0.01, N=6) during acute colitis.

A CB2R agonist inhibits colonic nociception during acute colitis, but not in healthy mice. A sub-analgesic combination of CB1R and MOR agonists can inhibit pain in healthy and inflamed mice, while combining sub-analgesic CB2R and MOR agonists is only inhibitory during acute colitis.

NRC