Objective To investigate the analgesia induced by cobrotoxin (CT) from venom of Naja naja atra, and the effects of atropine and naloxone on the antinociceptive activity of CT in rodent pain models. Methods CT was administered intraperitoneally (33.3,50,7μg/kg) , intra-cerebral venticularly (2.4μg /kg) or microinjected into periaque-ductal gray (PAG,1.2μg /kg). The antinociceptive action was tested using the hot-plate test and the acetic acid writhing test in mice and rats. The involvement of cholinergic system and the opioid system in CT-induced analgesia was examined by pretreatment of animals with atropine (0.5mg/kg, im or 10mg/kg, ip) or naloxone (3mg/kg,ip) . The effect of CT on motor activity was tested using the Animex test. Results CT (33.3,50 and 75μg/kg,ip) exhibited a dose-dependent analgesic action in mice as determined with hot-plate test and acetic acid writhing test. In the mouse acetic acid writhing test, the intra-cerebral ventricle administration of CT 2.4μg/kg (1/23th of a systemic dose) produced marked analgesic effects. Microinjection of CT 1.2μg/kg (1/46th of systemic dose) into the PAG also elicited a robust analgesic action in the hot-plate test in rats. Atropine at 0.5 mg/kg (im) or naloxone at 3 mg/kg (ip) failed to block the analgesic effects of CT, but atropine at 10 mg/kg (ip) did antagonize the analgesia mediated by CT in the mouse acetic acid writhing test. At the highest effective dose of antinociception (75μg/kg) , CT did not change the spontaneous mobility of mice. Conclusion These results suggest that CT from Naja naja atra venom has analgesic effects. Central nervous system may be involved in CT’ analgesic effects and the PAG may be the primary central site where CT exerts its effects. The central cholinergic system but not opioid system appears to be involved in the antinociceptive action of CT. Objective To investigate the analgesia induced by cobrotoxin (CT) from venom of Naja naja atra, and the effects of atropine and naloxone on the antinociceptive activity of CT in rodent pain models. Methods CT was administered intraperitoneally (33.3, 50, 7 μg / kg) , intra-cerebral venticularly (2.4 μg / kg) or microinjected into periaque-ductal gray (PAG, 1.2 μg / kg). The antinociceptive action was tested using the hot-plate test and the acetic acid writhing test in mice and rats. involvement of cholinergic system and the opioid system in CT-induced analgesia was examined by pretreatment of animals with atropine (0.5 mg / kg, im or 10 mg / kg, ip) or naloxone (3 mg / kg, ip) Results CT (33.3,50 and 75 μg / kg, ip) exhibited a dose-dependent analgesic action in mice as determined with hot-plate test and acetic acid writhing test. In the mouse acetic acid writhing test, the intra-cerebral ventricle administration of CT 2.4 μg / kg (1 Microinjection of CT 1.2 μg / kg (1/46 th of systemic dose) into the PAG also elicited a robust analgesic action in the hot-plate test in rats. Atropine at 0.5 mg / kg (im) or naloxone at 3 mg / kg (ip) failed to block the analgesic effects of CT, but atropine at 10 mg / kg (ip) did antagonize the analgesia mediated by CT in the mouse acetic acid writhing test. At the Most effective dose of antinociception (75 μg / kg), CT did not change the spontaneous mobility of mice. Conclusion These results suggest that CT from Naja naja atra venom has analgesic effects. Central nervous system may be involved in CT ’analgesic effects and the PAG The central cholinergic system but not opioid system appears to be involved in the antinociceptive action of CT.