Evaluation of the induction of vasoactive mediators from equine digital vein endothelial cells by endotoxin

Menzies-Gow, N J and Bailey, S R and Berhane, Y and Brooks, A C and Elliott, J (2008) Evaluation of the induction of vasoactive mediators from equine digital vein endothelial cells by endotoxin. AMERICAN JOURNAL OF VETERINARY RESEARCH, 69 (3). pp. 349-355.

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Abstract

Objective - To determine the effect of endotoxin (lipopolysaccharide [LPS]) on vasoactive mediator production by cultured equine digital vein endothelial cells (EDVECs). Sample Population - EDVECs obtained from forelimb digital veins of 7 healthy adult horses. Procedures - EDVECs were incubated with or without LPS (1 mu g/mL) for 0, 2, 4, 6, 22, and 24 hours. The EDVECs were incubated for 18 hours with LPS (10 mu g/mL to 1 mu g/mL) with or without ibuprofen, cycloheximide, or L-nitroarginine methyl ester. Medium concentrations of prostacyclin, cyclic guanosine monophosphate, endothelin-1, and thromboxane A(2) were determined. Changes in inducible nitric oxide synthase and cyclooxygenase-2 expression were determined. Results - LPS stimulated mean 4.2- and 14.1-fold increases in EDVEC prostacyclin and cyclic guanosine monophosphate production, respectively, after 22 hours. These effects were LPS concentration-dependent (LPS concentrations that induced a response halfway between the maximum response and baseline of 1.50 and 1.22 ng/mL, respectively). The LPS-induced cyclic guanosine monophosphate production was significantly inhibited (to basal concentrations) by L-nitroarginine methyl ester, and prostacyclin production was inhibited by cycloheximide and ibuprofen. Production of thromboxane A(2) by EDVECs was not detected. Endothelin-1 accumulated in the medium, but LPS did not enhance its production. Inducible nitric oxide synthase. expression in EDVECs was not detected with the available antibodies, whereas LPS stimulated cyclooxygenase-2 expression in a time- and concentration-dependent manner. Conclusions and Clinical Relevance - LPS stimulated vasoactive mediator production by equine endothelial cells, which may play a role in LPS-induced digital hypoperfusion.