Objectives To investigate the effects and involved mechanisms of scopolamine (Scop) on rabbit ear blood vessels. Methods Rabbit ear blood vessels were desympathetic and desensory innervation with surgical operation. Diameters of dorsal auricular arterial trunks in vivo were measured with a pair of compasses and the ruler in a dissecting microscope, and effluents from isolated ear under constant perfusion pressure were recorded with a digital drop-recorder. Results Intramuscular injection of Scop 0.1 mg/kg made the diameter of denerved dorsal auricular arterial trunks, as well as that of innerved ones, significantly increased. Scop by itself, at the maximal concentration (Cmax) of 3 μM, 30 μM and 300 μM, did not alter the effluent flow from the isolated denervated rabbit ear, but chlorpromazine (CPZ), at Cmax of 1 μM, acetylcholine (ACh), 0.25μM, all significantly increased the effluent flow, and norepinephrine (NE), 0.1μM, significantly decreased the effluent. Scop, 3 μM, did not affect ACh (0.25μM)-induced the increase of effluent flow, but Scop,30μM, alleviated the increase. Scop, 3μM, did not affect NE (0.1 μM)-induced the decrease of effluent flow, but Scop, 10, 30 and 100 μM, significantly alleviated the decrease. Conclusions The study suggests that Scop has no direct vasodilator effect. The vasodilator effect of Scop is not due to the blockade of muscarinic receptor. However, Scop can dilate blood vessels contracted by α1-adrenoceptor activation.