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First, aortic rings from control rats were relaxed by ACh, whereas those from ZDF rats were relaxed by sodium nitroprusside (SNP), and the effect of insulin was evaluated in both cases. Second, effects of insulin on the relaxation in aortic rings from control rats were assessed. Third, possible mechanisms were explored by testing the effects of specific inhibitors on these actions of insulin. Acetylcholine-induced relaxation was significantly reduced in aortic rings from ZDF rats. Insulin significantly potentiated the relaxation response to SNP in rings from control rats, whereas the response to acetylcholine was not significantly affected by insulin in the presence of glucose. The relaxation response to acetylcholine and that induced by insulin were similarly inhibited by the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester, whereas it was inhibited by the antioxidant aminoguanidine. 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