Serteyn, Didier[Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux >]
Mouithys-Mickalad, Ange[Université de Liège - ULg > > Centre de l'oxygène : Recherche et développement (C.O.R.D.) >]
Society for Free Radical International 16th biennal meeting
du 6 septembre 2012 au 9 septembre 2012
[en] Curcumin ; ROS ; Neutrophils ; HL-60 cells
[en] Neutrophils (PMNs) are involved in host defense against infections through the production of reactive oxygen species (ROS) and the release of an oxidant enzyme, myeloperoxidase (MPO), to kill pathologic agents. But, an excessive stimulation of PMNs is associated with development of inflammatory diseases. Neutrophils are prime targets to control inflammatory events and the therapeutic use of polyphenols is proposed to lower oxidative stress. The aim of this work was to study antioxidant effect of NDS27, a water-soluble form of curcumin, on stimulated equine PMNs and human promyelocytic leukemia cells (HL-60) which are less differenciated.
2',7'-Dichlorofluorescin diacetate and lucigenin were used to measure ROS production by activated HL-60 cells or PMNs. NDS27 (10-6 to 10-4 M) was pre-incubated with cells and eliminated before their activation to study its intracellular effects on ROS production. The effect of NDS27 on MPO activity released by the cells was determined by SIEFED. Likewise, the ability of NDS27 to enter into the cells was checked by HPLC on the cellular extracts.NDS27 significantly and dose-dependently inhibited the ROS production in both cell types without affecting their viability. Its intracellular effect showed higher efficiency for PMNs while its interaction with HL-60 cells remained better. The activity of MPO released by PMNs and HL-60 cells was decreased by NDS27 with a more efficient effect for PMNs. Our findings suggest that the greater efficiency of NDS27 in mature PMNs is not due to a better membrane permeability or a better interaction between membrane and NDS27, but rather to an inhibitory effect on the ROS production by the more mature cells, probably by targeting the enzymes implied in respiratory burst like MPO and NADPH oxidase. The modulatory effect of NDS27 towards oxidant activity of cells involved in immune and inflammatory response opens therapeutic perspectives to control pathologies with excessive inflammatory reactions.
Centre de l’Oxygène, Recherche et Développement - CORD
Fonds pour la formation à la Recherche dans l'Industrie et dans l'Agriculture (Communauté française de Belgique) - FRIA
Purification de la NADPH oxydase des neutrophiles équins : mise au point de nouveaux outils pour la quantification et la mesure spécifique de son activité et pour la recherche d'inhibiteurs de l'enzyme