ONO-1714, a new inducible nitric oxide synthase inhibitor, attenuates diaphragmatic dysfunction associated with cerulein-induced pancreatitis in rats.

OBJECTIVES: Acute experimental pancreatitis (induced by cerulein) recently has been reported to cause marked diaphragmatic dysfunction, which may contribute to respiratory distress in this setting. In cerulein-induced acute pancreatitis, expression of inducible nitric oxide synthase is induced to produce a large amount of nitric oxide. Nitric oxide excessively produced has been implicated in diaphragmatic dysfunction induced by a variety of etiologies. The aims of the current study were, first, to examine whether nitric oxide overproduced through inducible nitric oxide synthase is involved in cerulein-induced impairment of diaphragmatic function, and second, if demonstrated, to assess effects of ONO-1714, an inducible nitric oxide synthase inhibitor, on diaphragmatic dysfunction associated with cerulein-induced acute pancreatitis.

DESIGN: Prospective, randomized animal study.

SETTING: University research laboratory.

SUBJECTS: Ninety-one male Sprague-Dawley rats, weighing 200-250 g.

INTERVENTIONS: Rats were randomly divided into seven groups (n = 8 each): CONT-SAL, CAER-SAL, CONT-ONO, CAER-DEX, CAER-AMI, CAER-ONOhigh, and CAER-ONOlow. Groups labeled CAER received two consecutive intraperitoneal doses (50 microg/kg) of cerulein, whereas groups labeled CONT received two consecutive intraperitoneal injections of saline. Groups labeled SAL received intraperitoneal saline before cerulein or saline. The group labeled DEX received 2 mg/kg intraperitoneal dexamethasone, and the group labeled AMI received 100 mg/kg intraperitoneal aminoguanidine. The groups labeled ONO, ONOhigh, and ONOlow received ONO-1714 at 0.1 mg/kg, 0.1 mg/kg, and 0.03 mg/kg, respectively, before cerulein or saline.

MEASUREMENTS AND MAIN RESULTS: Diaphragmatic contractility and fatigability were assessed in vitro by using muscle strips excised from the costal diaphragms 6 hrs after the first dose of cerulein or saline. Expression of inducible nitric oxide synthase protein in the diaphragm was assessed by immunohistochemistry by using anti-inducible nitric oxide synthase antibody. Plasma concentrations of nitrite plus nitrate and diaphragmatic concentrations of malondialdehyde were measured. With another set of rats (n = 5 each group), diaphragmatic inducible nitric oxide synthase activity was determined. Twitch and tetanic tensions and tensions generated during fatigue trial were lower in group CAER-SAL than in group CONT-SAL. Cerulein increased diaphragmatic malondialdehyde and plasma nitrite plus nitrate concentrations. Positive immunostaining for inducible nitric oxide synthase protein was found in group CAER-SAL. Dexamethasone and aminoguanidine attenuated the diaphragmatic mechanical damages. A high dose of ONO-1714 attenuated cerulein-induced impairment of diaphragmatic contractility and endurance capacity, although a low dose of the drug failed to do so.

CONCLUSIONS: Cerulein-induced diaphragmatic dysfunction was attributable, in part, to nitric oxide overproduced via inducible nitric oxide synthase. Pretreatment with ONO-1714 at a dose of 0.1 mg/kg attenuated diaphragmatic dysfunction associated with cerulein-induced pancreatitis in rats assessed by contractile profiles and endurance capacity. This beneficial effect of ONO-1714 may be attributable, in part, to inhibition of diaphragmatic lipid peroxidation induced by nitric oxide-derived free radicals.