Arthritis & Rheumatism, Volume 63,
November 2011 Abstract Supplement
Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Chicago, Illinois November 4-9, 2011.
Superoxide Anion Mediates the L-Selectin Down-Regulation Induced by Non-Steroidal Anti-Inflammatory Drugs In Human Neutrophils.
Dominguez-Luis1, M. Jesus, Herrera-Garcia2, Ada, Arce-Franco3, M.Teresa, Cardenas4, Susana, Rodriguez-Pardo5, Marta, Feria5, Manuel, Sanchez-Madrid4, Francisco
Hospital Universitario de Canarias, La Laguna, Spain
Hospital Universitario de Canarias, La Laguna. Tenerife, Spain
Rheumatology Service, La Laguna, Spain
Hospital Universitario La Princesa, Madrid, Spain
University of La Laguna, La Laguna. Tenerife, Spain
A group of non-steroidal anti-inflammatory drugs (NSAIDs) are able to induce in neutrophils the shedding of L-selectin, a surface molecule that plays a critical step in the migration of leukocytes to site of inflammation, through a mechanism still not well-understood. The aim of this work was to study both the functional effect of NSAIDs on the neutrophils/endothelial cells dynamic interaction, and the potential implication of reactive oxygen species (ROS) production in the L-selectin down-regulation caused by NSAIDs
Human neutrophils were isolated from peripheral blood of healthy volunteers and patients with chronic granulomatous disease (CDG) (p47phox-/-), a group of hereditary diseases in which cells of the immune system have difficulty forming superoxide radical. The dynamic interaction between neutrophils and activated endothelium was studied in a flow chamber. Surface expression of L-selectin and CD11b were assayed by flow cytometry. Neutrophil-free supernatant L-selectin concentration was determined by ELISA. Intracellular and mitochondrial ROS production was detected by flow cytometry using dihidroethidium (DHE) and MitoSOX Red, respectively. Wilcoxon signed-rank test (p<0.05) was used to evaluate the statistical significance.
When human neutrophils were incubated with diclofenac but not with piroxicam, a significant reduction in the number of cells that rolled on activated endothelial cells was observed. Dithiol reducing agents such as, DL-Dithiothreitol (DTT) and 2,3-dimercapto-1-propane-sulfonic acid (DMPS) abrogated the down-regulatory effects of diclofenac, flufenamic acid and meclofenamic acid on neutrophils L-selectin basal expression. Several NSAIDs (n=10) caused variable increases in the mitochondrial and intracellular ROS concentration in neutrophils that were inversely proportional to the change they produced in L-selectin surface expression (r=-0.8, p< 0.01). The inhibition of NADPH oxidase activity with diphenyleneiodonium (DPI) prevented the down-regulation of L-selectin by diclofenac. The pre-incubation of neutrophils with superoxide dismutase but not with catalase showed both a significant protective effect on the L-selectin down-regulation induced by several NSAIDs and prevented diclofenac's effect on neutrophil rolling (p<0.05). Interestingly, neutrophils from CGD patients showed a significant less ability to release L-selectin from neutrophil surface than control in response to diclofenac.
All these data indicate that: 1) a group of NSAIDs is capable of interfering with the ability of neutrophils to interact with endothelial cells by means of the L-selectin-shedding and 2) the generation of superoxide anion by activation of NADPH oxidase at plasma membrane level seems to play a major role in the L-selectin shedding induced by NSAID in neutrophils, suggesting that ROS generation may assume a potentially novel function as regulators of inflammatory response
To cite this abstract, please use the following information:
Dominguez-Luis, M. Jesus, Herrera-Garcia, Ada, Arce-Franco, M.Teresa, Cardenas, Susana, Rodriguez-Pardo, Marta, Feria, Manuel, et al; Superoxide Anion Mediates the L-Selectin Down-Regulation Induced by Non-Steroidal Anti-Inflammatory Drugs In Human Neutrophils. [abstract]. Arthritis Rheum 2011;63 Suppl 10 :30