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1 School of Life Sciences, Napier University, Edinburgh, United Kingdom
2 ELEGI Laboratory, University of Edinburgh, Edinburgh, United Kingdom
* To whom correspondence should be addressed. E-mail: da.brown{at}napier.ac.uk.
We have previously examined the ability of air pollution particles (PM10) to promote release of the pro-inflammatory cytokine tumour necrosis factor alpha (TNF-
) from human peripheral blood mononuclear cells and demonstrated a role for calcium as a signalling molecule in this process. We have now studied the ability of oxidative stress induced by a synthetic oxidant tert-butylhydroperoxide (tBHP) to induce TNF production via calcium signalling in the mouse macrophage cell line (J774). The oxidant tBHP significantly increased intracellular calcium and the release of TNF- in J774 cells, an effect which was reduced to control levels by inhibition of calcium signalling with verapamil, BAPTA-AM and W7. This study also investigated interactions between PM10 treated macrophages and epithelial cells by using conditioned medium (CM) from PM10 treated mononuclear cells to stimulate the release of the neutrophil chemoattractant chemokine IL-8 from A549 lung epithelial cells. TNF- protein release was demonstrated in human mononuclear cells after PM10 treatment, an effect which was inhibited by calcium antagonists. Treatment of A549 cells with monocyte/PM10 CM produced increased IL-8 release which was reduced with CM from monocyte/PM10/calcium antagonist treatments. The expression of ICAM-1 was increased after incubation with CM from monocyte/PM10 treatment and this increase was prevented by treatment with CM from monocyte/PM10/calcium antagonist. These data demonstrate a link between oxidative stress, calcium and inflammatory mediator production in macrophages and lung epithelial cells.
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