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Competition for the amino acid arginine by endothelial nitric-oxide synthase (NOS3) and (pro-)inflammatory NO-synthase (NOS2) during endotoxemia appears essential in the derangement of the microcirculatory flow. This study investigated the role of NOS2 and NOS3 combined with/without citrulline supplementation on the NO-production and microcirculation during endotoxemia. Wildtype (C57BL6/N background; control;  n  = 36),  Nos2 -deficient, ( n  = 40),  Nos3 -deficient ( n  = 39) and  Nos2/Nos3 -deficient mice ( n  = 42) received a continuous intravenous LPS infusion alone (200 μg total, 18 h) or combined with L-citrulline (37.5 mg, last 6 h). The intestinal microcirculatory flow was measured by side-stream dark field (SDF)-imaging. The jejunal intracellular NO production was quantified by in vivo NO-spin trapping combined with electron spin-resonance (ESR) spectrometry. Amino-acid concentrations were measured by high-performance liquid chromatography (HPLC). LPS infusion decreased plasma arginine concentration in control and  Nos3 −/−   compared to  Nos2 −/−   mice. Jejunal NO production and the microcirculation were significantly decreased in control and  Nos2 −/−   mice after LPS infusion. No beneficial effects of L-citrulline supplementation on microcirculatory flow were found in  Nos3 −/−   or  Nos2 −/− /Nos3 −/−   mice. This study confirms that L-citrulline supplementation enhances de novo arginine synthesis and NO production in mice during endotoxemia with a functional NOS3-enzyme (control and  Nos2 −/−   mice), as this beneficial effect was absent in  Nos3 −/−   or  Nos2 −/− /Nos3 −/−   mice.

international journal of molecular sciences

Microcirculatory Function during Endotoxemia—A Functional Citrulline-Arginine-NO Pathway and NOS3 Complex Is Essential to Maintain the Microcirculation