Arginase attenuates inhibitory nonadrenergic noncholinergic nerve-induced nitric oxide generation and airway smooth muscle relaxation.
BACKGROUND
Recent evidence suggests that endogenous arginase activity potentiates airway responsiveness to methacholine by attenuation of agonist-induced nitric oxide (NO) production, presumably by competition with epithelial constitutive NO synthase for the common substrate, L-arginine. Using guinea pig tracheal open-ring preparations, we now investigated the involvement of arginase in the modulation of neuronal nitric oxide synthase (nNOS)-mediated relaxation induced by inhibitory nonadrenergic noncholinergic (iNANC) nerve stimulation.


METHODS
Electrical field stimulation (EFS; 150 mA, 4 ms, 4 s, 0.5 - 16 Hz)-induced relaxation was measured in tracheal preparations precontracted to 30% with histamine, in the presence of 1 microM atropine and 3 microM indomethacin. The contribution of NO to the EFS-induced relaxation was assessed by the nonselective NOS inhibitor L-NNA (0.1 mM), while the involvement of arginase activity in the regulation of EFS-induced NO production and relaxation was investigated by the effect of the specific arginase inhibitor nor-NOHA (10 microM). Furthermore, the role of substrate availability to nNOS in EFS-induced relaxation was measured in the presence of various concentrations of exogenous L-arginine.


RESULTS
EFS induced a frequency-dependent relaxation, ranging from 6.6 +/- 0.8% at 0.5 Hz to 74.6 +/- 1.2% at 16 Hz, which was inhibited with the NOS inhibitor L-NNA by 78.0 +/- 10.5% at 0.5 Hz to 26.7 +/- 7.7% at 8 Hz (P < 0.01 all). In contrast, the arginase inhibitor nor-NOHA increased EFS-induced relaxation by 3.3 +/- 1.2-fold at 0.5 Hz to 1.2 +/- 0.1-fold at 4 Hz (P < 0.05 all), which was reversed by L-NNA to the level of control airways in the presence of L-NNA (P < 0.01 all). Similar to nor-NOHA, exogenous L-arginine increased EFS-induced airway relaxation (P < 0.05 all).


CONCLUSIONS
The results indicate that endogenous arginase activity attenuates iNANC nerve-mediated airway relaxation by inhibition of NO generation, presumably by limiting L-arginine availability to nNOS.
Maarsingh
Harm
H
Department of Molecular Pharmacology, University Centre for Pharmacy, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, The Netherlands. h.maarsingh@rug.nl
Tio
Marieke A
MA
Zaagsma
Johan
J
Meurs
Herman
H
D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D001119:Arginase / Q000378:metabolism*; D004558:Electric Stimulation / Q000379:methods; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008297:Male; D009119:Muscle Contraction; D009126:Muscle Relaxation; D009130:Muscle, Smooth / Q000294:innervation* / Q000503:physiopathology*; D009433:Neural Inhibition; D009474:Neurons / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D011941:Receptors, Adrenergic / Q000378:metabolism; D011950:Receptors, Cholinergic / Q000378:metabolism; D014132:Trachea / Q000294:innervation* / Q000503:physiopathology*
D011941:Receptors, Adrenergic; D011950:Receptors, Cholinergic; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D001119:Arginase; D000109:Acetylcholine