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Neutrophil migration towards C 5a and CXCL 8 is prevented by non‐steroidal anti‐inflammatory drugs via inhibition of different pathways
Author(s) -
Bertolotto Maria,
Contini Paola,
Ottonello Luciano,
Pende Aldo,
Dallegri Franco,
Montecucco Fabrizio
Publication year - 2014
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1111/bph.12670
Subject(s) - chemistry , degranulation , pertussis toxin , chemotaxis , protein kinase b , pharmacology , microbiology and biotechnology , biochemistry , receptor , signal transduction , biology , g protein
Background and Purpose Non‐steroidal anti‐inflammatory drugs ( NSAID s) have been shown to induce PG‐independent anti‐inflammatory actions. Here, we investigated the role of three different NSAID s (naproxen, ibuprofen and oxaprozin) on neutrophil responses to CXCL 8 and C 5a. Experimental Approach Human neutrophils were isolated from healthy volunteers by dextran and F icoll‐ H ypaque density gradients. Neutrophils were pre‐incubated with different concentrations (1–100 µM) of NSAIDs or kinase inhibitors. Neutrophil degranulation into supernatants was tested by elisa and zymography. Neutrophil chemotaxis was determined using Boyden chambers. F ‐actin polymerization was determined by A lexa‐ F luor 488‐conjugated phalloidin fluorescent assay. Integrin expression was assessed by flow cytometry. The phosphorylation of intracellular kinases was studied by W estern blot. Key Results Pretreatment with NSAIDs did not affect neutrophil degranulation, but inhibited neutrophil migration and polymerization of F ‐actin, in response to CXCL 8 and C 5a. Pretreatment with different NSAID s prevented C 5a‐induced integrin ( CD 11b) up‐regulation, while only ibuprofen reduced CXCL 8‐induced CD 11b up‐regulation. Pre‐incubation with naproxen or oxaprozin, but not ibuprofen, inhibited the PI 3 K / A kt‐dependent chemotactic pathways. Both endogenous (released in cell supernatants) or exogenous (added to cell cultures) PGE 2 did not affect C 5a‐ or CXCL 8‐induced activities. Short‐term incubation with NSAID s did not affect neutrophil PGE 2 release. Conclusion and Implications Treatment with NSAIDs reduced C 5a‐ and CXCL 8‐induced neutrophil migration and F ‐actin polymerization via different mechanisms. Inhibition by ibuprofen was associated with integrin down‐regulation, while naproxen and oxaprozin blocked the PI 3 K / A kt pathway. Both NSAID actions were independent of COX inhibition and PGE 2 release.