title: Thymoquinone strongly inhibits fMLF-induced neutrophil functions and exhibits anti-inflammatory properties in vivo.
abstract: Polymorphonuclear neutrophils are key players in host defense against pathogens through the robust production of superoxide anion by the NADPH oxidase and the release of antibacterial proteins from granules. However, inappropriate release of these agents in the extracellular environment induces severe tissue injury, thereby contributing to the physiopathology of acute and chronic inflammatory disorders. Many studies have been carried out to identify molecules capable of inhibiting phagocyte functions, in particular superoxide anion production, for therapeutic purposes. In the present study, we show that thymoquinone (TQ), the major component of the volatile oil from Nigella sativa (black cumin) seeds strongly inhibits fMLF-induced superoxide production and granules exocytosis in neutrophils. The inhibition of superoxide anion was not due to a scavenger effect, as TQ did not inhibit superoxide anion produced by the xanthine/xanthine oxidase system. Interestingly, TQ impaired the phosphorylation on Ser-304 and Ser-328 of p47(PHOX), a cytosolic subunit of the NADPH oxidase. TQ also attenuated specific and azurophilic granule exocytosis in fMLF-stimulated neutrophils as evidenced by decreased cell surface expression of gp91(PHOX) and CD11b, and release of myeloperoxidase. Furthermore, both the PKC and MAPK pathways, which are involved in p47(PHOX) phosphorylation and granules exocytosis, respectively, were inhibited by TQ in fMLF-stimulated neutrophils. Finally, in a model of pleurisy induced by λ-carrageenan in rats, TQ reduced neutrophil accumulation in the pleural space, showing that it not only inhibits PMN functions in vitro, but also exhibits anti-inflammatory properties in vivo. Thus, TQ possesses promising anti-inflammatory therapeutic potential.
authors.affiliation: INSERM U1149, ERL 8252 CNRS, Centre de Recherche sur l'Inflammation, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Laboratoire d'Excellence Inflamex, Faculté de Médecine, Site Xavier Bichat, Paris, France; Laboratoire de Chimie des Matériaux et des Vivants: Activité et réactivité, Faculté des Sciences, Université El-Hadj Lakhdar, Batna, Algeria; Laboratoire de Biochimie appliquée-Inflammation: Activités Pharmaco-Biologiques des Substances Naturelles, Département de Biochimie, Université Ferhat Abbes, Sétif, Algeria.
authors.forename: Kaouthar
authors.initials: K
authors.lastname: Boudiaf
authors.affiliation: INSERM U1149, ERL 8252 CNRS, Centre de Recherche sur l'Inflammation, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Laboratoire d'Excellence Inflamex, Faculté de Médecine, Site Xavier Bichat, Paris, France.
authors.forename: Margarita
authors.initials: M
authors.lastname: Hurtado-Nedelec
authors.affiliation: INSERM U1149, ERL 8252 CNRS, Centre de Recherche sur l'Inflammation, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Laboratoire d'Excellence Inflamex, Faculté de Médecine, Site Xavier Bichat, Paris, France.
authors.forename: Sahra Amel
authors.initials: SA
authors.lastname: Belambri
authors.affiliation: INSERM U1149, ERL 8252 CNRS, Centre de Recherche sur l'Inflammation, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Laboratoire d'Excellence Inflamex, Faculté de Médecine, Site Xavier Bichat, Paris, France.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Marie
authors.affiliation: Département des Sciences de la Nature et de la Vie, Université Mohamed Khider, Biskra, Algeria.
authors.forename: Yacine
authors.initials: Y
authors.lastname: Derradji
authors.affiliation: Laboratoire de Biochimie appliquée-Inflammation: Activités Pharmaco-Biologiques des Substances Naturelles, Département de Biochimie, Université Ferhat Abbes, Sétif, Algeria.
authors.forename: Mustapha
authors.initials: M
authors.lastname: Benboubetra
authors.affiliation: INSERM U1149, ERL 8252 CNRS, Centre de Recherche sur l'Inflammation, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Laboratoire d'Excellence Inflamex, Faculté de Médecine, Site Xavier Bichat, Paris, France.
authors.forename: Jamel
authors.initials: J
authors.lastname: El-Benna
authors.affiliation: INSERM U1149, ERL 8252 CNRS, Centre de Recherche sur l'Inflammation, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Laboratoire d'Excellence Inflamex, Faculté de Médecine, Site Xavier Bichat, Paris, France. Electronic address: my-chan.dang@inserm.fr.
authors.forename: Pham My-Chan
authors.initials: PM
authors.lastname: Dang
mesh_terms: D000328:Adult; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D016227:Benzoquinones / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D009249:NADP / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D031881:Nigella sativa / Q000737:chemistry*; D010998:Pleurisy / Q000188:drug therapy / Q000276:immunology / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D013481:Superoxides / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D016227:Benzoquinones; D013481:Superoxides; D009249:NADP; D009240:N-Formylmethionine Leucyl-Phenylalanine; C003466:thymoquinone; D002118:Calcium
keywords: Degranulation; NADPH oxidase; Neutrophils; Phosphorylation; ROS; Superoxide anion; Thymoquinone; p47(PHOX)