Neutrophil migration towards C5a and CXCL8 is prevented by non‐steroidal anti‐inflammatory drugs via inhibition of different pathways

Abstract

BACKGROUND AND PURPOSENon-steroidal anti-inflammatory drugs (NSAIDs) have been shown to induce PG-independent anti-inflammatory actions. Here, we investigated the role of three different NSAIDs (naproxen, ibuprofen and oxaprozin) on neutrophil responses to CXCL8 and C5a. EXPERIMENTAL APPROACHHuman neutrophils were isolated from healthy volunteers by dextran and Ficoll-Hypaque 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 Alexa-Fluor 488-conjugated phalloidin fluorescent assay. Integrin expression was assessed by flow cytometry. The phosphorylation of intracellular kinases was studied by Western blot. KEY RESULTSPretreatment with NSAIDs did not affect neutrophil degranulation, but inhibited neutrophil migration and polymerization of F-actin, in response to CXCL8 and C5a. Pretreatment with different NSAIDs prevented C5a-induced integrin (CD11b) up-regulation, while only ibuprofen reduced CXCL8-induced CD11b up-regulation. Pre-incubation with naproxen or oxaprozin, but not ibuprofen, inhibited the PI3K/Akt-dependent chemotactic pathways. Both endogenous (released in cell supernatants) or exogenous (added to cell cultures) PGE2 did not affect C5a-or CXCL8-induced activities. Short-term incubation with NSAIDs did not affect neutrophil PGE2 release. CONCLUSION AND IMPLICATIONSTreatment with NSAIDs reduced C5a-and CXCL8-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 PI3K/Akt pathway. Both NSAID actions were independent of COX inhibition and PGE2 release.

Phosphoinositide-3 kinase gamma regulates caspase-1 activation and leukocyte recruitment in acute murine gout

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AQP5-1364A/C polymorphism and the AQP5 expression influence sepsis survival and immune cell migration: a prospective laboratory and patient study

“…The lower compartment contained 10 −8 M fMLP (Sigma-Aldrich, Taufkirchen, Germany) in 500 µl RPMI or control media. The cells were incubated at 37 °C with 5% CO 2 in air for 0.5 and 1.5 h in duplicate for each sample as described before [21]. The migrated cells were counted after 0.5 and 1.5 h using the MUSE Count & Viability Kit (Merck Millipore, Darmstadt, Germany).…”

Host-pathogen interplay in the respiratory environment of cystic fibrosis

“…High dose ibuprofen decreases neutrophil migration as measured through oral mucosal washes in clinical trials [87] and slows the progression of CF lung disease, particularly in children [88]. The mechanism involved in ibuprofen-affected neutrophil migration has not been clearly delineated but may be due to reduced CD11 upregulation and polymerization of actin in response to IL-8 and C5a [89]. Azithromycin has been shown to reduce pulmonary exacerbations and improve CF lung function [90] by different mechanisms, which include suppression of cytokine production, increased tight junctions and protection of respiratory epithelium from damage.…”