P Bodin1, G Burnstock. 1. Autonomic Neuroscience Institute, Royal Free Hospital School of Medicine, London, UK.
Abstract
OBJECTIVE AND DESIGN: The effects of lipopolysaccharide (LPS), a potent inflammatory mediator, on the shear stress stimulated release of adenosine triphosphate (ATP) were investigated on endothelial cells from human umbilical vein in primary culture. METHODS: Human umbilical vein endothelial cells (HUVEC) in primary cultures were subjected to shear stress using a cone and plate apparatus. ATP released by the cells was measured by luminometry, using a luciferin-luciferase assay. RESULTS: Under conditions of shear stress alone (25dyn/cm2), ATP accumulates into the culture medium and reaches a maximum after 3 to 5 min of stimulation (121.7+/-13.2 pmol/ml). The shear stress-stimulated release of ATP was significantly increased after a 4 h pre-incubation of endothelial cells with 50 microg/ml (314.4+/-26.7 pmol/ml) and 10microg/ml lipopolysaccharide (207.7+/-22.2 pmol/ml). Dexamethasone, an anti-inflammatory glucocorticoid, inhibited the effects of lipopolysaccharide. CONCLUSIONS: These results show that non-damaged endothelial cells release ATP under experimental inflammatory conditions and support an early role of extracellular ATP in the inflammatory process.
OBJECTIVE AND DESIGN: The effects of lipopolysaccharide (LPS), a potent inflammatory mediator, on the shear stress stimulated release of adenosine triphosphate (ATP) were investigated on endothelial cells from human umbilical vein in primary culture. METHODS:Human umbilical vein endothelial cells (HUVEC) in primary cultures were subjected to shear stress using a cone and plate apparatus. ATP released by the cells was measured by luminometry, using a luciferin-luciferase assay. RESULTS: Under conditions of shear stress alone (25dyn/cm2), ATP accumulates into the culture medium and reaches a maximum after 3 to 5 min of stimulation (121.7+/-13.2 pmol/ml). The shear stress-stimulated release of ATP was significantly increased after a 4 h pre-incubation of endothelial cells with 50 microg/ml (314.4+/-26.7 pmol/ml) and 10microg/ml lipopolysaccharide (207.7+/-22.2 pmol/ml). Dexamethasone, an anti-inflammatory glucocorticoid, inhibited the effects of lipopolysaccharide. CONCLUSIONS: These results show that non-damaged endothelial cells release ATP under experimental inflammatory conditions and support an early role of extracellular ATP in the inflammatory process.
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