Abstract: The aim of present study was to investigate the role of opioid and α-adrenergic receptors in antispasmodic activity of Ruta chalepensis (L.) hydroalcoholic extract (RCHE) in rat ileum contractions induced by KCl. A piece of ileum (2 cm) was dissected out from male Wistar rats and mounted in an organ bath containing air bubbled Tyrode solution (37°C). Isotonic Transducer is exceptionally sensitive and stable and connected to Harvard Universal Oscillograph (UK). The tissue was under 1 g initial tension and contractions were recorded isotonically. The RCHE was prepared by macerated with alcohol (70%). Ileum was contracted by KCl (60 mM) and cumulative concentrations of RCHE were applied (0.01-0.07 mg mL-1) before and after tissue incubation (30 min) with naloxon (1 μM) or phentolamine (1 μM). RCHE reduced KCl-induced contraction in a concentration dependent manner (p<0.001). However, tissue incubation with naloxone and phentolamine attenuated the spasmolytic effect of RCHE (p<0.001). We conclude that in the antispasmodic activity of RCHE, at least in part, the opioid and α -adrenergic receptors are involved.