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The effects of the ethanol extract of Tilia cordata Miller (Tiliaceae) were studied in vitro using intestinal smooth muscle cells of guinea pigs dispersed by collagenase. The extract induced a dose-dependent contraction of the dispersed smooth muscle cells. The obtained data indicated a direct effect of the T. cordata extract on the intestinal smooth muscle cells. In addition, the contractions induced by the extract were inhibited by atropine. These observations indicate involvement of some active constituents, with cholinergic properties, found in the alcohol extract to induce contraction of the intestinal smooth muscle cells via activation of the muscarinic receptors.