Context: Alternanthera repens (L.) Kuntze (Amaranthaceae) is widely used in Mexican traditional medicine for the treatment of gastrointestinal disorders that are mainly related to diarrhea. Objective: The aim of the present study was to investigate the spasmolytic effect of hexane (Hx), methanol (Me) and aqueous (Aq) extracts as well as chromatographic Me fractions (F1-F6) of A. repens in rat ileum. Materials and methods: Dried and powdered aerial parts were used to obtain the extracts. The rat ileum preparations were incubated in Tyrode's solution gassed (95% O 2 -5% CO 2 ) at 37 C. The effect on the contractile response of isolated ileum was evaluated by obtaining cumulative concentration-response curves to CaCl 2 , KCl, 5-HT and acetylcholine in the absence and presence of different doses of Aq (0.56-2.09 mg/mL), Me (0.24-0.91 mg/mL) and Hx (0.24-0.91 mg/mL) extracts, as well as six Me fractions of 0.66 mg/mL (F1 to F6).