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  • Fluid resuscitation therapy in endotoxemic hamsters improves survival and attenuates capillary perfusion deficits and inflammatory responses by a mechanism related to nitric oxide.

Fluid resuscitation therapy in endotoxemic hamsters improves survival and attenuates capillary perfusion deficits and inflammatory responses by a mechanism related to nitric oxide.

Journal of translational medicine (2014-08-26)
Nivaldo Ribeiro Villela, Ana Olimpia Maia Teixeira dos Santos, Marcos Lopes de Miranda, Eliete Bouskela
ABSTRACT

Relative hypovolemia is frequently found in early stages of severe sepsis and septic shock and prompt and aggressive fluid therapy has become standard of care improving tissue perfusion and patient outcome. This paper investigates the role of the nitric oxide pathway on beneficial microcirculatory effects of fluid resuscitation. After skinfold chamber implantation procedures and endotoxemia induction by intravenous Escherichia coli lipopolysaccharide administration (2 mg x kg(-1)), male golden Syrian hamsters were fluid resuscitated and then sequentially treated with L-Nω-Nitroarginine and L-Arginine hydrochloride (LPS/FR/LNNA group). Intravital microscopy of skinfold chamber preparations allowed quantitative analysis of microvascular variables including venular leukocyte rolling and adhesion. Macro-hemodynamic, biochemical and hematological parameters as well as survival rate were also evaluated. Endotoxemic hamsters treated with fluid therapy alone (LPS/FR group) and non-treated animals (LPS group) served as controls. Fluid resuscitation was effective in reducing lipopolysaccharide-induced microcirculatory changes. After 3 hours of lipopolysaccharide administration, non-fluid resuscitated animals (LPS group) had the lowest functional capillary density (1% from baseline for LPS group vs. 19% for LPS/FR one; p <0.05). At the same time point, arteriolar mean internal diameter was significantly wider in LPS/FR group than in LPS one (100% vs. 50% from baseline). Fluid resuscitation also reduced leukocyte-endothelium interactions and sequestration (p <0.05 for LPS vs. LPS/FR group) and increased survival (median survival time: 2 and 5.5 days for LPS and LPS/FR groups, respectively; p <0.05). Nitric oxide synthase inhibition prevented these protective effects, while L-Arginine administration markedly restored many of them. Our results suggest that the underlying mechanism of fluid therapy is the restoration of nitric oxide bioavailability, because inhibition of NOS prevented many of its beneficial effects. Nevertheless, further investigations are required in experimental models closer to conditions of human sepsis to confirm these results.

MATERIALS
Product Number
Brand
Product Description

Supelco
Rhodamine 6G, analytical standard
Sigma-Aldrich
Rhodamine 6G, Dye content 99 %
Sigma-Aldrich
Rhodamine 6G, Dye content ~95 %
Sigma-Aldrich
Rhodamine 6G, suitable for fluorescence, BioReagent
SAFC
L-Arginine monohydrochloride
Arginine hydrochloride, European Pharmacopoeia (EP) Reference Standard
Supelco
Arginine Hydrochloride, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
L-Arginine monohydrochloride, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
L-Arginine monohydrochloride, not synthetic, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Arginine monohydrochloride, reagent grade, ≥98% (HPLC), powder
Sigma-Aldrich
Nω-Nitro-L-arginine, ≥98% (TLC)
Supelco
L-Arginine hydrochloride solution, 100 mM amino acid in 0.1 M HCl, analytical standard
Sigma-Aldrich
L-Arginine monohydrochloride, BioUltra, ≥99.5% (AT)