Tachykinins stimulate nitric oxide generation by canine cultured tracheal epithelium

We studied the effects of tachykinins on the generation of nitric oxide (NO) from canine cultured tracheal epithelial cells using a specific amperometric sensor for this molecule. Immersion of the NO-selective electrode in the medium bathing the cells detected the baseline current of 30.5-61.7 pA, which corresponded to NO concentration ([NO]) at 44.0 +/- 7.6 nM (mean +/- S.E.M.). Substance P (SP, 10(-6) M) increased the current from 51.3 +/- 9.8 to 73.6 +/- 11.4 pA (P < 0.001), an effect that was not affected by NG-nitro-D-arginine methylester, but inhibited by NG-nitro-L-arginine methylester by 83 +/- 9% (P < 0.001), and this inhibition was restored by the subsequent addition of L-arginine, but not by D-arginine. SP and neurokinin A (NKA) increased [NO] in a dose-dependent manner, the maximal increases from the baseline level being 71.0 +/- 14.9 and 33.4 +/- 8.5 nM, respectively (P < 0.001 for each), whereas neurokinin B (NKB) had no effect. In the presence of phosphoramidon, the response of each tachykinin was augmented, but the rank order of potency was still NKA > SP >> NKB. These results suggest that NO is spontaneously released from airway epithelium and that tachykinins stimulate epithelial NO generation via NK2 receptors.