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The Functional Significance of the Rho/Rho-Kinase Pathway in Human Erythrocytes
Objective: Erythrocyte deformability, which can be influenced by various intracellular signaling mechanisms, such as nitric oxide, cAMP, cGMP, and protein kinases, is the most important physiological factor providing the blood flow in microcirculation. However, the functional significance of the Rho...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Galenos Publishing
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4102045/ https://www.ncbi.nlm.nih.gov/pubmed/25035675 http://dx.doi.org/10.4274/tjh.2013.0115 |
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author | Tiftik, R. Nalan Başkurt, Oğuz K. Kul, Seval Büyükafşar, Kansu |
author_facet | Tiftik, R. Nalan Başkurt, Oğuz K. Kul, Seval Büyükafşar, Kansu |
author_sort | Tiftik, R. Nalan |
collection | PubMed |
description | Objective: Erythrocyte deformability, which can be influenced by various intracellular signaling mechanisms, such as nitric oxide, cAMP, cGMP, and protein kinases, is the most important physiological factor providing the blood flow in microcirculation. However, the functional significance of the Rho/Rho-kinase pathway, which contributes cell shape changes and the reorganization of the actin cytoskeleton, has yet to be explored in erythrocytes. Therefore, we examined the influence of several activators and inhibitors of Rho/Rho-kinase signaling on human erythrocyte deformability. Materials and Methods: RhoA and ROCK-2 proteins were studied by western blotting. Influences of 2 Rho-kinase inhibitors, fasudil and Y-27632 (both 10-7 to 10-4 M), on erythrocyte deformability was determined by ektacytometer at various shear stresses (0-30 Pa) in the presence or absence of a known Rho activator, lysophosphatidic acid (LPA, 10-5 to 5x10-5 M, 1-15 min). Results: LPA incubation reduced deformability with concomitant RhoA-GTP inhibition. Y-27632 and fasudil also decreased deformability, but had no effect on LPA-induced reduction of deformability. Rho inhibitor C3 had no effect on RhoA activation. Reduction in RhoA activation was induced by sub-hemolytic mechanical stress. Conclusion: Our findings may indicate that the Rho/Rho-kinase pathway could contribute to the regulation of deformability of human erythrocytes. |
format | Online Article Text |
id | pubmed-4102045 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Galenos Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-41020452014-07-17 The Functional Significance of the Rho/Rho-Kinase Pathway in Human Erythrocytes Tiftik, R. Nalan Başkurt, Oğuz K. Kul, Seval Büyükafşar, Kansu Turk J Haematol Research Article Objective: Erythrocyte deformability, which can be influenced by various intracellular signaling mechanisms, such as nitric oxide, cAMP, cGMP, and protein kinases, is the most important physiological factor providing the blood flow in microcirculation. However, the functional significance of the Rho/Rho-kinase pathway, which contributes cell shape changes and the reorganization of the actin cytoskeleton, has yet to be explored in erythrocytes. Therefore, we examined the influence of several activators and inhibitors of Rho/Rho-kinase signaling on human erythrocyte deformability. Materials and Methods: RhoA and ROCK-2 proteins were studied by western blotting. Influences of 2 Rho-kinase inhibitors, fasudil and Y-27632 (both 10-7 to 10-4 M), on erythrocyte deformability was determined by ektacytometer at various shear stresses (0-30 Pa) in the presence or absence of a known Rho activator, lysophosphatidic acid (LPA, 10-5 to 5x10-5 M, 1-15 min). Results: LPA incubation reduced deformability with concomitant RhoA-GTP inhibition. Y-27632 and fasudil also decreased deformability, but had no effect on LPA-induced reduction of deformability. Rho inhibitor C3 had no effect on RhoA activation. Reduction in RhoA activation was induced by sub-hemolytic mechanical stress. Conclusion: Our findings may indicate that the Rho/Rho-kinase pathway could contribute to the regulation of deformability of human erythrocytes. Galenos Publishing 2014-06 2014-06-10 /pmc/articles/PMC4102045/ /pubmed/25035675 http://dx.doi.org/10.4274/tjh.2013.0115 Text en © Turkish Journal of Hematology, Published by Galenos Publishing. http://creativecommons.org/licenses/by/2.5/ 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 work is properly cited. |
spellingShingle | Research Article Tiftik, R. Nalan Başkurt, Oğuz K. Kul, Seval Büyükafşar, Kansu The Functional Significance of the Rho/Rho-Kinase Pathway in Human Erythrocytes |
title | The Functional Significance of the Rho/Rho-Kinase Pathway in Human Erythrocytes |
title_full | The Functional Significance of the Rho/Rho-Kinase Pathway in Human Erythrocytes |
title_fullStr | The Functional Significance of the Rho/Rho-Kinase Pathway in Human Erythrocytes |
title_full_unstemmed | The Functional Significance of the Rho/Rho-Kinase Pathway in Human Erythrocytes |
title_short | The Functional Significance of the Rho/Rho-Kinase Pathway in Human Erythrocytes |
title_sort | functional significance of the rho/rho-kinase pathway in human erythrocytes |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4102045/ https://www.ncbi.nlm.nih.gov/pubmed/25035675 http://dx.doi.org/10.4274/tjh.2013.0115 |
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