Al-Ani, Bahjat, Hewett, Peter W., Ahmed, Suborna, Cudmore, Melissa, Fujisawa, Takeshi, Ahmad, Shakil and Ahmed, Asif (2006). The release of nitric oxide from S-nitrosothiols promotes angiogenesis. PLoS ONE, 1 (1),
Abstract
Free nitric oxide (NO) reacts with sulphydryl residues to form S-nitrosothiols, which act as NO reservoirs. We sought to determine whether thiol-preserving agents and antioxidants, such as dithiothreitol (DTT) and vitamin C, induce NO release from S-nitrosylated proteins in endothelial cell cultures to promote angiogenesis. NO release was measured directly in cell supernatants using a Sievers NO Analyser, and in vitro angiogenesis was assessed by quantifying capillary-like tube network formation of porcine aortic endothelial cells (PAEC) on growth factor-reduced Matrigel. Incubation of PAEC with DTT or vitamin C significantly increased NO release in a concentration-dependent manner. However, the nitric oxide synthase (NOS) inhibitors, L-NNA and L-NIO, had no effect on DTT- or vitamin C-induced NO release, and there was no concomitant increase in the phosphorylation of endothelial NOS at serine-1177 following DTT or vitamin C treatment. DTT and vitamin C increased capillary-like tube network formation by nine- and two-fold, respectively, and the addition of copper ions doubled the effect of vitamin C. Surprisingly, DTT maintained endothelial tube networks for up to one month under serum-free conditions, and selective inhibitors of guanylyl cyclase (ODQ) and PKG (KT-5823) blocked this, demonstrating the requirement of cyclic GMP and PKG in this process. Both DTT and vitamin C are capable of releasing sufficient NO from S-nitrosothiols to induce capillary morphogenesis. This study provides the first evidence that increased denitrosylation leads to increased bioavailability of NO, independent of NOS activity, to promote sustained angiogenesis.
Publication DOI: | https://doi.org/10.1371/journal.pone.0000025 |
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Divisions: | College of Health & Life Sciences > School of Biosciences College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine College of Health & Life Sciences > Aston Medical School College of Health & Life Sciences |
Additional Information: | © 2006 Al-Ani et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
Uncontrolled Keywords: | angiogenesis,antioxidants,cell membrane,endothelial cells,growth facotrs,nitiric acid,phosphorylation,vitamin c |
Last Modified: | 10 Oct 2024 07:12 |
Date Deposited: | 15 May 2014 15:40 |
Full Text Link: |
http://www.plos ... al.pone.0000025 |
Related URLs: | PURE Output Type: | Article |
Published Date: | 2006-12-20 |
Authors: |
Al-Ani, Bahjat
Hewett, Peter W. Ahmed, Suborna Cudmore, Melissa Fujisawa, Takeshi Ahmad, Shakil ( 0000-0002-9294-0475) Ahmed, Asif ( 0000-0002-8755-8546) |