AMINOGUANIDINE ATTENUATES THE DELAYED CIRCULATORY FAILURE AND IMPROVES SURVIVAL IN RODENT MODELS OF ENDOTOXIC-SHOCK

1 We have investigated the effects of aminoguanidine, a relatively selective inhibitor of the cytokine-inducible isoform of nitric oxide synthase (iNOS), on the delayed circulatory failure, vascular hyporeactivity to vasoconstrictor agents, and iNOS activity in a rat model of circulatory shock induced by bacterial endotoxin (E. coli lipopolysaccharide; LPS). In addition, we have evaluated the effect of aminoguanidine on the 24 h survival rate in a murine model of endotoxaemia. 2 Male Wistar rats were anaesthetized and instrumented for the measurement of mean arterial blood pressure (MAP) and heart rate (HR). Injection of LPS (10 mg kg(-1), i.v.) resulted in a fall in MAP from 115 +/- 4 mmHg (time 0, control) to 79 +/- 9 mmHg at 180 min (P<0.05, n = 10). The presser effect of noradrenaline (NA, 1 mu g kg(-1), i.v.) was also significantly reduced at 60, 120 and 180 min after LPS injection. In contrast, animals pretreated with aminoguanidine (15 mg kg(-1), i.v., 20 min prior to LPS injection) maintained a significantly higher MAP (at 180 min, 102 +/- 3 mmHg, n = 10, P<0.05) when compared to rats given only LPS (LPS-rats). Cumulative administration of aminoguanidine (15 mg kg(-1) and 45 mg kg(-1)) given 180 min after LPS caused a dose-related increase in MAP and reversed the hypotension. Aminoguanidine also significantly alleviated the reduction of the presser response to NA: indeed, at 180 min, the presser response returned to normal in aminoguanidine pretreated LPS-rats. 3 Thoracic aortae obtained from rats at 180 min after LPS showed a significant reduction in the contractile responses elicited by NA (10(-9)-10(-6) M). Pretreatment with aminoguanidine (15 mg kg(-1), i.v., at 20 min prior to LPS) significantly prevented this LPS-induced hyporeactivity to NA ex vivo. 4 Endotoxaemia for 180 min resulted in a significant increase in iNOS activity in the lung from 0.6 +/- 0.2 pmol mg(-1) min(-1) (control, n = 4) to 4.8 +/- 0.3 pmol mg(-1) min(-1) (P<0.05, n = 6). In LPS-rats treated with aminoguanidine, iNOS activity in the lung was attenuated by 44 +/- 5% (n = 6, P<0.05). Moreover, when added in vitro to lung homogenates obtained from LPS-rats, aminoguanidine and N-omega-nitro-L-arginine methyl ester (L-NAME; 10(-8) to 10(-3) M) caused a concentration-dependent inhibition of iNOS activity (n = 3-6, IC50: 30 +/- 12 and 11 +/- 6 mu M, respectively P>0.05). In contrast, aminoguanidine was a less potent inhibitor than L-NAME of the constitutive nitric oxide synthase in rat brain homogenates (n = 3-6, IC50 is 140 +/- 10 and 0.6 +/- 0.1 mu M, respectively, P<0.05). In addition, the inhibitory effect of aminoguanidine on iNOS activity showed a slower onset than that of L-NAME (maximal inhibition at 90 min and 30 min, respectively). 5 Treatment of conscious Swiss albino (T/O) mice with a high dose of endotoxin (60 mg kg(-1), i.p.) resulted in a survival rate of only 8% at 24 h (n = 12). However, therapeutic application of aminoguanidine (15 mg kg(-1), i.p. at 2 h and 6 h after LPS) increased the 24 h survival rate to 75% (n = 8), whereas L-NAME (3 mg kg(-1), i.p. at 2 h and 6 h after LPS) did not affect the survival rate (11%, n = 9). 6 Thus, aminoguanidine inhibits iNOS activity and attenuates the delayed circulatory failure caused by endotoxic shock in the rat and improves survival in a murine model of endotoxaemia. Aminoguanidine, or novel, more potent selective inhibitors of iNOS may be useful in the therapy of septic shock.