S Kopprasch1, A Gatzweiler, M Kohl, H E Schröder. 1. Technical University of Dresden, Carl Gustav Carus Medical School, Department of Internal Medicine III, Germany.
Abstract
UNLABELLED: The in vitro effect of endothelin-1 (ET-1) on the capacity of polymorphonuclear leukocytes (PMNLs) to generate reactive oxygen species (ROS) was investigated. Human PMNLs were separated from healthy volunteers and preincubated for 10 min. at 37 degrees C with varying concentrations (10(-7)-10(-12) M) of ET-1. After subsequent stimulation with FMLP (10(-7) M) or opsonized zymosan (0.5 mg/ml) the intra- and extracellular generation of ROS was assessed by luminol-amplified chemiluminescence, superoxide radical (.O2-) and hydrogen peroxide (H2O2) production. RESULTS: ET-1 alone failed to stimulate ROS generation. Neither the capacity for extracellular generation of oxygen metabolites nor the production of ROS with an intracellular origin was changed after preincubation of PMNLs with ET-1. ET-1 did not cause a shift of the .O2-/H2O2 production ratio after stimulation of PMNLs with FMLP. These findings suggest that ET-1 in vitro does not prime human PMNLs for enhanced production of ROS.
UNLABELLED: The in vitro effect of endothelin-1 (ET-1) on the capacity of polymorphonuclear leukocytes (PMNLs) to generate reactive oxygen species (ROS) was investigated. Human PMNLs were separated from healthy volunteers and preincubated for 10 min. at 37 degrees C with varying concentrations (10(-7)-10(-12) M) of ET-1. After subsequent stimulation with FMLP (10(-7) M) or opsonized zymosan (0.5 mg/ml) the intra- and extracellular generation of ROS was assessed by luminol-amplified chemiluminescence, superoxide radical (.O2-) and hydrogen peroxide (H2O2) production. RESULTS:ET-1 alone failed to stimulate ROS generation. Neither the capacity for extracellular generation of oxygen metabolites nor the production of ROS with an intracellular origin was changed after preincubation of PMNLs with ET-1. ET-1 did not cause a shift of the .O2-/H2O2 production ratio after stimulation of PMNLs with FMLP. These findings suggest that ET-1 in vitro does not prime human PMNLs for enhanced production of ROS.
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