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Šantak, Borislav; Radermacher, Peter; Iber, Thomas; Adler, Jens; Wachter, Ulrich; Vassilev, Damian; Georgieff, Michael; Vogt, Josef
British journal of pharmacology, December 1997, Letnik: 122, Številka: 8Journal Article
1 In this investigation the NO production rate is quantified in the pig during normotensive endotoxin‐induced shock with increased cardiac output and during subsequent treatment with the NO synthase inhibitor Nω‐monomethy‐l‐arginine (l‐NMMA). NO production rate was derived from the plasma isotope‐enrichment of 15N‐labelled nitrate (15NO3−). 2 Three groups of animals (control, n=5; endotoxin, n=6; endotoxin+l‐NMMA, n=6) were anaesthetized and instrumented for the measurement of systemic and pulmonary haemodynamics. Each animal received a primed‐continuous infusion of stable, non‐radioactively labelled Na15NO3 (bolus 30 mg, infusion rate 2.1 mg h−1). Arterial blood samples were taken 5, 10, 15, 30, 60 and 90 min later and every 90 minutes until the end of the experiment. 3 Continuous i.v. infusion of endotoxin was incrementally adjusted until mean pulmonary artery pressure (PAP) reached 50 mmHg and subsequently titrated to keep mean PAP ∼amp;35 mmHg. Hydroxyethylstarch was administered as required to maintain mean arterial pressure (MAP)>60 mmHg. Six hours after the start of the endotoxin continuous i.v. l‐NMMA (1 mg kg−1 h−1) was administered to the endotoxin+l‐NMMA group. Haemodynamic data were measured before as well as 9 h after the start of the endotoxin. 4 After conversion of NO3− to nitro‐trimethoxybenzene and gas chromatography‐mass spectrometry analysis the total NO3− pool, basal NO3− production rate and the increase per unit time in NO3− production rate were calculated from the time‐course of the 15NO3− plasma isotope‐enrichment. A two compartment model was assumed for the NO3− kinetics, one being an active pool in which newly generated NO3− appears and from which it is eliminated, the other being an inactive volume of distribution in which only passive exchange takes place with the active compartment. 5 Although MAP did not change during endotoxin infusion alone, cardiac output (CO) increased by 42±40% (P<0.05 versus baseline) by the end of the experiment due to a significant (P<0.05 versus baseline) fall in systemic vascular resistance (SVR) to 65±25% of the baseline value. l‐NMMA given with endotoxin did not change MAP, and both CO and SVR were maintained close to the pre‐shock levels. 6 Baseline plasma NO3− concentrations were 43±13 and 40±10 μmol l−1 in the control and endotoxin animals, respectively, and did not differ at the end of the experiment (39±8 and 44±15 μmol l−1, respectively). The mean NO3− pool and basal NO3− production rate were 1155±294 μmol and 140±32 μmol h−1, respectively, without any intergroup difference. Endotoxin significantly increased NO3− production rate (23±10 μmol h−2, P<0.05 versus control (6±7 μmol h−2) and endotoxin+l‐NMMA groups). l‐NMMA given with endotoxin (−1±2 μmol h−2, P<0.05 versus control and endotoxin groups) had no effect. 7 Analysis of the time course of the 15NO3− plasma isotope enrichment during primed‐continuous infusion of Na15NO3 allowed us to quantify the endotoxin‐induced increase in NO3− production rate independently of total NO3− plasma concentrations. Low‐dose l‐NMMA blunted the increase in NO3− production rate while maintaining basal NO3− formation.
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