RESUMO
1. Electrical field stimulation (EFS) of intrinsic nerves in the rat proximal duodenum induces a frequency-dependent non-adrenergic-non-cholinergic (NANC) relaxation response. 2. The inhibitors of L-arginine-NO synthase L-NG-nitro arginine methyl-ester (L-NAME) and L-NOARG (L-NG-nitro arginine) reduced the NANC relaxations elicited by EFS in a dose- and time-dependent manner; L-NOARG was two times more potent than L-NAME (IC50 = 14.3 vs 25.2 microM) and these effects were partially reverted by the addition of 300-1000 microM L-arginine but not of 300-1000 microMD-arginine. Relaxation caused by vasoactive intestinal peptide (VIP; 0.1 microM) or ATP (20 microM) was not blocked by L-NAME or L-NOARG. 3. The magnitude of the blockade caused by L-NAME and L-NOARG was dependent on the frequency of stimulation. At low frequencies (below 1 Hz) both L-NAME and L-NOARG abolished the relaxations, while at 2 to 8 Hz only partial inhibition was observed (maximal inhibition: 44.6 per cent +/- 5.2 and 63.4 per cent +/- 3.4, respectively) 4. The basal tonus of the duodenum was increased by 10-300 microM L-NAME and 10-300 microM L-NOARG and this effect was blocked by 1 mM L-arginine. 5. Nitric oxide generated from acidified NaNO2 caused a dose-dependent (EC50 = 2.75 microM) relaxation of the duodenum which was not affected by 100 microM L-NAME, 100 microM L-NOARG or 1 microM tetrodotoxin (TTX). 6. NADPH-diaphorase positive neurons and fibers identified by histochemistry were present in the myenteric plexus and along both circular and longitudinal muscle fibers indicating that nitric oxide could be synthetized by these neural structures