PMID- 28017872
OWN - NLM
STAT- MEDLINE
DCOM- 20180705
LR  - 20181202
IS  - 1089-8611 (Electronic)
IS  - 1089-8603 (Linking)
VI  - 63
DP  - 2017 Feb 28
TI  - Nitrite-derived nitric oxide reduces hypoxia-inducible factor 1alpha-mediated 
      extracellular vesicle production by endothelial cells.
PG  - 1-12
LID - S1089-8603(16)30182-3 [pii]
LID - 10.1016/j.niox.2016.12.005 [doi]
AB  - INTRODUCTION: Extracellular vesicles (EVs) are small, spherical particles 
      enclosed by a phospholipid bilayer ( approximately 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 (NaNO(2)) 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% O(2), 
      24 h) and compared to endothelial cells exposed to normoxia (21% O(2)) with and 
      without the presence of sodium nitrite (NaNO(2)) (30 muM). Allopurinol (100 muM), 
      an inhibitor of xanthine oxidoreductase, was added both alone and in combination 
      with NaNO(2) 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-1alpha 
      and HIF-2alpha 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% O(2)) 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 
      (NaNO(2)) reduced the hypoxic enhancement of EV production. Treatment of cells 
      with the xanthine oxidoreductase inhibitor allopurinol, in addition to NaNO(2) 
      attenuated the NaNO(2)-attributed suppression of hypoxia-mediated EV release. 
      Transfection of cells with HIF-1alpha siRNA, but not HIF-2alpha 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-1alpha, but not HIF-2alpha. Furthermore, the reduction of 
      NO(2)(-) to NO via xanthine oxidoreductase during hypoxia appears to inhibit 
      HIF-1alpha-mediated EV production.
CI  - Copyright (c) 2016 Elsevier Inc. All rights reserved.
FAU - Burnley-Hall, Nicholas
AU  - Burnley-Hall N
AD  - School of Medicine, Cardiff University, Cardiff, CF24 4HQ, UK.
FAU - Willis, Gareth
AU  - Willis G
AD  - Division Newborn Medicine, Boston Children's Hospital, Harvard Medical School, 
      Harvard University, Boston, 02115, MA, USA.
FAU - Davis, Jessica
AU  - Davis J
AD  - Institute of Cancer & Genetics, Cardiff University, Cardiff, CF14 4XN, UK.
FAU - Rees, D Aled
AU  - Rees DA
AD  - Neurosciences and Mental Health Research Institute, School of Medicine, Cardiff 
      University, Cardiff, CF24 4HQ, UK.
FAU - James, Philip E
AU  - James PE
AD  - Cardiff School of Health Sciences, Cardiff Metropolitan University, Cardiff, CF5 
      2SG, UK. Electronic address: PJames@cardiffmet.ac.uk.
LA  - eng
PT  - Journal Article
DEP - 20161223
PL  - United States
TA  - Nitric Oxide
JT  - Nitric oxide : biology and chemistry
JID - 9709307
RN  - 0 (Basic Helix-Loop-Helix Transcription Factors)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Hypoxia-Inducible Factor 1, alpha Subunit)
RN  - 1B37H0967P (endothelial PAS domain-containing protein 1)
RN  - 31C4KY9ESH (Nitric Oxide)
RN  - 63CZ7GJN5I (Allopurinol)
RN  - EC 1.17.1.4 (Xanthine Dehydrogenase)
RN  - J06Y7MXW4D (Deferoxamine)
RN  - M0KG633D4F (Sodium Nitrite)
SB  - IM
MH  - Allopurinol/pharmacology
MH  - Basic Helix-Loop-Helix Transcription Factors/metabolism
MH  - Deferoxamine/pharmacology
MH  - Endothelial Cells/*metabolism/pathology
MH  - Enzyme Inhibitors/pharmacology
MH  - Extracellular Vesicles/chemistry/*metabolism
MH  - Human Umbilical Vein Endothelial Cells/metabolism
MH  - Humans
MH  - Hypoxia/metabolism
MH  - Hypoxia-Inducible Factor 1, alpha Subunit/*metabolism
MH  - Nitric Oxide/*physiology
MH  - Particle Size
MH  - Sodium Nitrite/metabolism
MH  - Xanthine Dehydrogenase/antagonists & inhibitors
OTO - NOTNLM
OT  - Extracellular vesicles
OT  - Hypoxia
OT  - Hypoxia-inducible factor
OT  - Nitric oxide
OT  - Nitrite
EDAT- 2016/12/27 06:00
MHDA- 2018/07/06 06:00
CRDT- 2016/12/27 06:00
PHST- 2016/10/04 00:00 [received]
PHST- 2016/12/07 00:00 [revised]
PHST- 2016/12/15 00:00 [accepted]
PHST- 2016/12/27 06:00 [pubmed]
PHST- 2018/07/06 06:00 [medline]
PHST- 2016/12/27 06:00 [entrez]
AID - S1089-8603(16)30182-3 [pii]
AID - 10.1016/j.niox.2016.12.005 [doi]
PST - ppublish
SO  - Nitric Oxide. 2017 Feb 28;63:1-12. doi: 10.1016/j.niox.2016.12.005. Epub 2016 Dec 
      23.