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Time course of nitric oxide production after endotoxin challenge in mice

¹Division of Nutrition and Metabolism, University Hospital Clementino Fraga Filho, Federal University of Rio de Janeiro, 21941-590 Rio de Janeiro, Brazil; and ²Department of Surgery, Maastricht University, NL-6200 MD Maastricht, The Netherlands

Submitted 1 December 2003 ; accepted in final form 15 July 2004

Nitric oxide (NO) regulates numerous processes during endotoxemia and inflammation. However, the sequential changes in whole body (Wb) nitric oxide (NO) production during endotoxemia in vivo remain to be clarified. Male Swiss mice were injected intraperitoneally with saline (control group) or lipopolysaccharide (LPS group). After 0, 2, 4, 6, 9, 12, and 24 h, animals received a primed constant infusion of L-[guanidino-¹⁵N₂-²H₂]arginine, L-[ureido-¹⁵N]citrulline, L-[5-¹⁵N]glutamine, and L-[ring-²H₅]phenylalanine in the jugular vein. Arterial blood was collected for plasma arginine (Arg), citrulline (Cit), glutamine (Gln), and phenylalanine (Phe) concentrations and tracer-to-tracee ratios. NO production was calculated as plasma Arg-to-Cit flux, Wb de novo Arg synthesis as plasma Cit-to-Arg flux, and Wb protein breakdown as plasma Phe flux. LPS reduced plasma Arg and Cit and increased Gln and Phe concentrations. Two peaks of NO production were observed at 4 and 12 h after LPS. Although LPS did not affect total Arg production, de novo Arg production decreased after 12 h. The second peak of NO production coincided with increased Wb Cit, Gln, and Phe production. In conclusion, the curve of NO production in both early and late phases of endotoxemia is not related to plasma Arg kinetics. However, because Wb Cit, Gln, and Phe fluxes increased concomitantly with the second peak of NO production, NO production is probably related to the catabolic phase of endotoxemia.

lipopolysaccharide; arginine; citrulline; glutamine; phenylalanine

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