[en] In neutrophils (PMNs), superoxide anion (O2●-), the first reactive oxygen species (ROS) produced to kill pathogenic agents, is generated by NADPH oxidase, an enzymatic complex formed by the translocation of cytosolic subunits to the membrane flavocytochrome b558. In horses, excessive activation of PMNs is often associated with deadly pathologies and the modulation of their ROS production by acting on NADPH oxidase is a prime target to manage inflammation. We developed a cell-free assay to measure the activity of equine NADPH oxidase assembled in vitro, in order to test the effects of natural or synthetic compounds on the enzyme activity or assembly. The cell-free assay was validated with diphenyleneiodonium chloride and Gp91ds-tat, two inhibitors largely described for human NADPH oxidase. The anti-oxidant effects of curcumin and resveratrol at final concentration ranging from 10-4 to 10-6 M were studied on whole cells by chemiluminescence (CL) and by cell-free assay, in which the molecule was added before or after the enzyme assembly. The CL assay demonstrated that curcumin efficiently inhibited the O2●- production and easily entered into PMNs or interacted with their membrane. Cell-free assay showed that curcumin acted on the reconstitution of NADPH oxidase even at 10-5 M, while resveratrol appeared to be an O2●- scavenger rather than an inhibitor of NADPH oxidase activity, since it acted from outside the cell in CL and after the complex assembly in cell-free assay. By acting directly on NADPH oxidase, curcumin should be a good candidate for the treatment of acute or inflammatory diseases involving an excessive ROS production.
Research center :
CORD - Centre de l'Oxygène, Recherche et Développement - ULiège
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Derochette, Sandrine ; Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Franck, Thierry ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Mouithys-Mickalad, Ange ; Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Ceusters, Justine ; Université de Liège - ULiège > Clinique des grands animaux (chirurgie)
Deby-Dupont, Ginette
Lejeune, Jean-Philippe
Neven, Philippe
Serteyn, Didier ; Université de Liège - ULiège > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux
Language :
English
Title :
Curcumin and resveratrol act by different ways on NADPH oxidase activity and reactive oxygen species produced by equine neutrophils
Alternative titles :
[fr] La curcumine et le resveratrol agissent de façon différente sur l'activité de la NADPH oxydase et les ROS produites par les neutrophiles équins
Publication date :
2013
Journal title :
Chemico-Biological Interactions
ISSN :
0009-2797
eISSN :
1872-7786
Publisher :
Elsevier, Limerick, Ireland
Volume :
206
Pages :
186-193
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
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
Funders :
FRIA - Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture [BE]
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