Rats with experimental colitis suffer from impaired gastric emptying. We previously showed that this phenomenon involves afferent neurons within the pelvic nerve. In this study, we aimed to identify the mediators involved in this afferent hyperactivation. Colitis was induced using trinitrobenzene sulphate instillation (TNBS). We determined gastric emptying (GE), distal front and geometric center (GC) of intestinal transit 30 min after intragastric administration of a semiliquid Evans blue solution. We evaluated the effects of the transient receptor potential vanilloid type 1 (TRPV1) antagonists capsazepine (5-10 mg/kg) and BCTC (1-10 mg/kg) and the calcitonin gene-related peptide (CGRP) receptor antagonist CGRP (8-37) (150 µg/kg). To determine TRPV1 receptor antagonist sensitivity, we examined their effect on capsaicin-induced relaxations of isolated gastric fundus muscle strips. Immunocytochemical staining of TRPV1 and RT-PCR analysis of TRPV1 mRNA were performed in dorsal root ganglion (DRG) L6-S1. TNBS-induced colitis reduced GE but had no effect on intestinal motility. Capsazepine reduced GE in controls but had no effect in rats with colitis. At doses which had no effects in controls, BCTC and CGRP (8-37) significantly improved colitis-induced gastroparesis. Capsazepine inhibited capsaicin-induced relaxations by 35% while BCTC completely abolished them. TNBS-induced colitis increased TRPV1-like immunoreactivity and TRPV1 mRNA content in pelvic afferent neuronal cell bodies in DRG L6-S1. In conclusion, distal colitis in rats impairs gastric emptying via sensitized pelvic afferent neurons. We provided pharmacological, immunocytochemical and molecular biological evidence that this sensitization is mediated by TRPV1 receptors and involves CGRP release.