Background: The petal′s hydro-alcoholic extract of Rosa damascena Mill. on ileum contractions of Wistar rats and its possible mechanism were investigated. Methods: Forty-eight male Wistar rats were divided into six groups. Ileum was placed adjacent to propranolol (1 μM), naloxone (1 μM) and L-NAME (100 μM) and also under the influence of different doses (2-8 mM) of calcium chloride. Results: Cumulative extract of R. damascena Mill. (100, 500, and 1000 mg/L) decreased ileum contractions induced by KCl (60 mM) in a dose-dependent manner (P < 0.0001). Propranolol and naloxone significantly decreased the inhibitory effect of the extract on contractions induced by KCl (P < 0.001), but L-NAME was ineffective. Furthermore, calcium led to the contraction of depolarized tissue through KCI and this contractile effect decreased significantly induced by the cumulative concentrations of the extract (P < 0.001). Conclusions: The results indicate that R. damascena Mill. dose-dependently (100, 500, and 1000 mg/L) decreases ileum movements of the rat probably through stimulating the β-adrenergic and opioid receptors and voltage-dependent calcium channels, and it may be used to treat digestive disorders.