Transient receptor potential vanilloid 1 (TRPV1) channels have been implicated in pain mechanisms and, particularly, in the development of hyperalgesia. We used selective TRPV1 antagonists (NGV-1, SB-750364 and JYL 1421) to assess the role of TRPV1 channels in repetitive noxious colorectal distension (CRD)-induced visceral pain responses in rats. Isobaric CRD (80 mmHg) induced a viscerosomatic response, indicative of visceral pain associated to the distension procedure. Repetition (12 consecutive distensions) of the CRD resulted in an increase in the response over time (119 ± 23% increase at distension 12, P < 0.05 vs response during the 1st distension) indicative of acute mechanical sensitization. NGV-1 (0.1, 0.3, 1 or 3 μmol/kg, i.v.) prevented in a dose-related manner the development of sensitization, without inducing hypoalgesic responses. SB-750364 (30 μmol/kg, i.v.) had a transitory effect, partially reducing the sensitization response, while JYL 1421 (4.7 μmol/kg, i.v.) was without effect. In the same conditions, the cannabinoid receptor 1 (CB 1) agonist, WIN55,212-2 (0.1 μmol/kg) reduced pain responses leading to a hypoalgesic state. At 3 μmol/kg, NGV-1, did not affect the pressure–volume relationship during CRD, indicating that TRPV1 channels do not modulate colonic compliance. These observations suggest that TRPV1 channels are involved in the development of acute mechanical colonic hyperalgesia during repetitive noxious CRD in rats. Antagonism of TRPV1 channels might result in antihyperalgesic effects without hypoalgesic activity and might be beneficial in the treatment of visceral pain disorders, such as irritable bowel syndrome. These observations warrant the clinical assessment of TRPV1 antagonists for the treatment of visceral pain.