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This study aimed to compare the effects of these mediators on pain, anxiety, and depression. Ninety\u2010one adult male Sprague\u2013Dawley rats were used for the experiments. Locomotor activity, elevated plus maze, forced swimming, tail clip, hot plate, and writhing tests were used for the assessments after the administration of <jats:sc>l<\/jats:sc>\u2010arginine (30\u2013100\u2009mg\/kg), a NO donor; NaHS (5\u201310\u2009mg\/kg), a H<jats:sub>2<\/jats:sub>S donor; and CORM\u20102 (5\u201310\u2009mg\/kg), CO donor. Intraperitoneal H<jats:sub>2<\/jats:sub>S, NO, malondialdehyde (MDA), glutathione (GSH), and tumor necrosis factor\u2010\u03b1 (TNF\u2010\u03b1) levels were determined by enzyme\u2010linked immunosorbent assay (ELISA). No statistical significance was found in the locomotor activity. NO and CO significantly extended latency at high doses in tail clip test. No significant activity was observed at any dose of all three substances on a hot plate. Both doses of CO and high doses of NO and H<jats:sub>2<\/jats:sub>S showed an antinociceptive effect in the writhing test. While the opioidergic system plays a role in the spinal antinociceptive effect of <jats:sc>l<\/jats:sc>\u2010arginine, both serotonergic and opioidergic systems play a role in its peripheral antinociceptive effect. The serotonergic system plays a role in the peripheral antinociceptive effect of CORM\u20102. The time spent in open arm increased significantly in all groups an elevated plus maze. High doses of all three substances significantly increased the duration of immobility in the forced swimming test. No statistical significance was observed in MDA, GSH, and TNF\u2010\u03b1 levels. High doses of NO and CO showed a spinal antinociceptive effect. Both doses of CO and high doses of NO and H<jats:sub>2<\/jats:sub>S showed a peripheral antinociceptive effect. 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