Show simple item record

dc.contributor.authorBurnley-Hall, Nicholas
dc.contributor.authorWillis, Gareth
dc.contributor.authorDavis, Jessica
dc.contributor.authorRees, D. Aled
dc.contributor.authorJames, Philip
dc.date.accessioned2017-01-24T13:45:01Z
dc.date.available2017-01-24T13:45:01Z
dc.date.issued2016-12-23
dc.identifier.citationBurnley-Hall, N., Willis, G., Davis, J., Rees, D.A. and James, P.E. (2017) 'Nitrite-derived nitric oxide reduces hypoxia-inducible factor 1α-mediated extracellular vesicle production by endothelial cells', Nitric Oxide, 63, pp.1-12.en_US
dc.identifier.issn1089-8603
dc.identifier.urihttp://hdl.handle.net/10369/8314
dc.descriptionThis article was published in Nitric Oxide on 23 December 2016 (online), available at http://dx.doi.org/10.1016/j.niox.2016.12.005en_US
dc.description.abstractIntroduction Extracellular vesicles (EVs) are small, spherical particles enclosed by a phospholipid bilayer (∼30–1000 nm) released from multiple cell types, and have been shown to have pathophysiological roles in a plethora of disease states. The transcription factor hypoxia-inducible factor-1 (HIF-1) allows for adaptation of cellular physiology in hypoxia and may permit the enhanced release of EVs under such conditions. Nitric oxide (NO) plays a pivotal role in vascular homeostasis, and can modulate the cellular response to hypoxia by preventing HIF-1 accumulation. We aimed to selectively target HIF-1 via sodium nitrite (NaNO2) addition, and examine the effect on endothelial EV, size, concentration and function, and delineate the role of HIF-1 in EV biogenesis. Methods Endothelial (HECV) cells were exposed to hypoxic conditions (1% O2, 24 h) and compared to endothelial cells exposed to normoxia (21% O2) with and without the presence of sodium nitrite (NaNO2) (30 μM). Allopurinol (100 μM), an inhibitor of xanthine oxidoreductase, was added both alone and in combination with NaNO2 to cells exposed to hypoxia. EV and cell preparations were quantified by nanoparticle tracking analysis and confirmed by electron microscopy. Western blotting and siRNA were used to confirm the role of HIF-1α and HIF-2α in EV biogenesis. Flow cytometry and time-resolved fluorescence were used to assess the surface and intravesicular protein content. Results Endothelial (HECV) cells exposed to hypoxia (1% O2) produced higher levels of EVs compared to cells exposed to normoxia. This increase was confirmed using the hypoxia-mimetic agent desferrioxamine. Treatment of cells with sodium nitrite (NaNO2) reduced the hypoxic enhancement of EV production. Treatment of cells with the xanthine oxidoreductase inhibitor allopurinol, in addition to NaNO2 attenuated the NaNO2-attributed suppression of hypoxia-mediated EV release. Transfection of cells with HIF-1α siRNA, but not HIF-2α siRNA, prior to hypoxic exposure prevented the enhancement of EV release. Conclusion These data provide evidence that hypoxia enhances the release of EVs in endothelial cells, and that this is mediated by HIF-1α, but not HIF-2α. Furthermore, the reduction of NO2− to NO via xanthine oxidoreductase during hypoxia appears to inhibit HIF-1α-mediated EV production.en_US
dc.language.isoenen_US
dc.publisherElsevieren_US
dc.relation.ispartofseriesNitric Oxide;
dc.subjectextracellular vesiclesen_US
dc.subjectHypoxiaen_US
dc.subjectHypoxia-inducible factoren_US
dc.subjectNitriteen_US
dc.subjectNitric oxideen_US
dc.titleNitrite-derived nitric oxide reduces hypoxia-inducible factor 1α-mediated extracellular vesicle production by endothelial cellsen_US
dc.typeArticleen_US
dc.identifier.doihttp://dx.doi.org/10.1016/j.niox.2016.12.005
dcterms.dateAccepted2016-12-15
rioxxterms.funderCardiff Metropolitan Universityen_US
rioxxterms.identifier.projectCardiff Metropolian (Internal)en_US
rioxxterms.versionAMen_US
rioxxterms.licenseref.urihttps://creativecommons.org/licenses/by-nc-nd/3.0/en_US
rioxxterms.licenseref.startdate2017-01-24
rioxxterms.freetoread.startdate2017-12-23
rioxxterms.funder.project37baf166-7129-4cd4-b6a1-507454d1372een_US


Files in this item

Thumbnail

This item appears in the following collection(s)

Show simple item record