A new synthetic flavone derivative, 6,7-dimethoxy-8-methyl-3 ,4 ,5-trihydroxyflavone, was studied for its capacity to protect the acetylcholine-induced relaxation of rabbit ear and cerebral arteries from inhibition by superoxide anion. This property was evaluated via two types of in vitro experiments, using rabbit ear or basilar arteries mounted in organ baths equipped for isometric tension measurement. When a high level of superoxide anion was generated by adding 3 10 - 4 M pyrogallol to the bath, the relaxation to acetylcholine was substantially inhibited. This inhibition was significantly reversed by both superoxide dismutase (25 and/or 50 U/mL) and the flavonoid (3 10 - 6 M and/or 10 - 5 M) in both types of arteries. In the presence of the basal level of superoxide anion, the responses to acetylcholine were significantly potentiated by the flavonoid (10 - 5 M) in the ear but not the basilar artery. Thus this flavonoid protects endothelium-dependent relaxation from high levels of superoxide anion possibly by scavenging superoxide anion and may have a certain therapeutic value as an agent capable of promoting natural vasodilatation.