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F2-isoprostanes affect macrophage migration and CSF-1 signalling
J M Proudfoot1, M W Murrey2, S McLean2
1School of Biomedical Sciences, University of Western Australia, Royal Perth Hospital Medical Research Foundation, GPO Box X2213, Perth, Western Australia 6847, Australia.
Free Radical Biology & Medicine
|August 11, 2018
Summary
15-F2t-isoprostane (15-F2t-IsoP) stimulates macrophage migration towards CSF-1, despite inhibiting cell adhesion. This isoprostane impacts macrophage signaling pathways, influencing their role in atherosclerosis development.
Area of Science:
- Biochemistry and Molecular Biology
- Cardiovascular Research
- Cell Biology
Background:
- Oxidative stress, indicated by F2-isoprostanes (F2-IsoP), is linked to atherosclerosis.
- Macrophages are key players in atherosclerosis, and their migration is crucial.
- Colony stimulating factor-1 (CSF-1) is vital for macrophage function and present in atherosclerotic plaques.
Purpose of the Study:
- To investigate the effect of 15-F2t-isoprostane (15-F2t-IsoP) on macrophage migration.
- To explore the underlying signaling pathways involved in 15-F2t-IsoP-mediated macrophage responses.
- To determine the role of specific adhesion kinases in 15-F2t-IsoP's effects on macrophages.
Main Methods:
- Macrophage cell lines (BAC1.2F5) and primary bone marrow-derived macrophages were used.
- Cell migration was assessed using Boyden chambers and wound healing assays.
- Signaling pathways were analyzed via Western blotting, and gene silencing (siRNA) was employed for FAK and Pyk2.
Main Results:
- 15-F2t-IsoP dose-dependently inhibited BAC1.2F5 macrophage spreading and adhesion but stimulated migration towards CSF-1.
- 15-F2t-IsoP decreased Akt and Rictor phosphorylation while increasing FAK, Pyk2, and paxillin phosphorylation.
- Pyk2 silencing counteracted 15-F2t-IsoP-induced reduction in cell area and phospho-paxillin adhesions, with a trend towards reduced migration.
Conclusions:
- 15-F2t-IsoP modulates macrophage adhesion and migration, impacting key signaling pathways.
- The findings highlight a novel role for 15-F2t-IsoP in macrophage behavior relevant to atherosclerosis.
- Pyk2 appears to play a significant role in mediating 15-F2t-IsoP's effects on macrophage adhesion and migration.
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