F 2 -isoprostanes affect macrophage migration and CSF-1 signalling.

Autor: Proudfoot JM; School of Biomedical Sciences, University of Western Australia, Royal Perth Hospital Medical Research Foundation, GPO Box X2213, Perth, Western Australia 6847, Australia. Electronic address: julie.proudfoot@uwa.edu.au., Murrey MW; School of Biomedical Sciences, University of Western Australia, The Queen Elizabeth II Medical Centre, Nedlands, Western Australia 6009, Australia. Electronic address: michael.murrey@research.uwa.edu.au., McLean S; School of Biomedical Sciences, University of Western Australia, The Queen Elizabeth II Medical Centre, Nedlands, Western Australia 6009, Australia. Electronic address: samantha.mclean@telethonkids.org.au., Greenland EL; School of Biomedical Sciences, University of Western Australia, The Queen Elizabeth II Medical Centre, Nedlands, Western Australia 6009, Australia. Electronic address: eloise.greenland@research.uwa.edu.au., Barden AE; Medical School, University of Western Australia, Royal Perth Hospital Medical Research Foundation, GPO Box X2213, Perth, Western Australia 6847, Australia. Electronic address: anne.barden@uwa.edu.au., Croft KD; School of Biomedical Sciences, University of Western Australia, Royal Perth Hospital Medical Research Foundation, GPO Box X2213, Perth, Western Australia 6847, Australia. Electronic address: kevin.croft@uwa.edu.au., Galano JM; Institut des Biomolécules Max Mousseron (IBMM), UMR 5247, CNRS, University Montpellier, ENSCM, Faculté de Pharmacie, 15 Av. Ch. Flahault, BP 14491, F-34093 Montpellier Cedex 05, France. Electronic address: Jean-Marie.Galano@umontpellier.fr., Durand T; Institut des Biomolécules Max Mousseron (IBMM), UMR 5247, CNRS, University Montpellier, ENSCM, Faculté de Pharmacie, 15 Av. Ch. Flahault, BP 14491, F-34093 Montpellier Cedex 05, France. Electronic address: Thierry.Durand@umontpellier.fr., Mori TA; Medical School, University of Western Australia, Royal Perth Hospital Medical Research Foundation, GPO Box X2213, Perth, Western Australia 6847, Australia. Electronic address: trevor.mori@uwa.edu.au., Pixley FJ; School of Biomedical Sciences, University of Western Australia, The Queen Elizabeth II Medical Centre, Nedlands, Western Australia 6009, Australia. Electronic address: fiona.pixley@uwa.edu.au.
Jazyk: angličtina
Zdroj: Free radical biology & medicine [Free Radic Biol Med] 2018 Oct; Vol. 126, pp. 142-152. Date of Electronic Publication: 2018 Aug 08.
DOI: 10.1016/j.freeradbiomed.2018.08.007
Abstrakt: F 2 -isoprostanes (F 2 -IsoP) are formed in vivo via free radical peroxidation of arachidonic acid. Enhanced oxidative stress is implicated in the development of atherosclerosis in humans and F 2 -IsoP have been detected in atherosclerotic plaque. Colony stimulating factor-1 (CSF-1) is essential to macrophage survival, proliferation and differentiation and has been detected in human atherosclerotic plaques. Accumulation of macrophages within the vascular wall is an important component of atherosclerosis but little is known about the effect of F 2 -IsoP on the migration of these cells. Our aim was to examine the effect of free and lipid-bound 15-F 2t -isoprostane (15-F 2t -IsoP) on macrophage migration and investigate the signalling pathways involved. Mouse macrophages (cell line BAC1.2F5) were pre-incubated with 15-F 2t -IsoP (free, bound to cholesterol or monoacylglycerol or within oxidized phospholipid) and cell migration was assessed using chemotaxis towards CSF-1 in Boyden chambers. Migration was also measured using the wound healing assay with primary mouse bone marrow derived macrophages. We showed that 15-F 2t -IsoP dose-dependently inhibited BAC1.2F5 macrophage spreading and adhesion but stimulated their migration towards CSF-1, with maximum effect at 10 µM. Analysis of CSF-1 stimulated signalling pathways in BAC1.2F5 macrophages showed that phosphorylation of Akt, a key mediator of cell migration, and one of its regulators, the mTORC2 component, Rictor, was significantly decreased. In contrast, phosphorylation of the adhesion kinases, FAK and Pyk2, and the adhesion scaffold protein, paxillin, was enhanced after treatment with 15-F 2t -IsoP. Mouse bone marrow macrophages were transfected with FAK or Pyk2 small interfering RNA (siRNA) to examine the role of FAK and Pyk2 in 15-F 2t -IsoP signalling. Pyk2 silencing inhibited 15-F 2t -IsoP-induced reduction in cell area and phospho-paxillin adhesion numbers. The size distribution of adhesions in the presence of 15-F 2t -IsoP was also affected by Pyk2 silencing and there was a trend for Pyk2 silencing to reduce 15-F 2t -IsoP-stimulated macrophage migration. These results demonstrate that 15-F 2t -IsoP affects macrophage adhesions and migration, which are integral components of macrophage involvement in atherosclerosis.
(Copyright © 2018 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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