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The Rab-Rabphilin system in injured human podocytes stressed by glucose overload and angiotensin II

Kidney injury in hypertension and diabetes entails, among in other structures, damage in a key cell of the glomerular filtration barrier, the podocyte. Podocytes are polarized and highly differentiated cells in which vesicular transport, partly driven by Rab GTPases, is a relevant process. The aim o...

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Autores principales: Martinez-Arroyo, Olga, Ortega, Ana, Perez-Hernandez, Javier, Chaves, Felipe J., Redon, Josep, Cortes, Raquel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Physiological Society 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7473899/
https://www.ncbi.nlm.nih.gov/pubmed/32567349
http://dx.doi.org/10.1152/ajprenal.00077.2020
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author Martinez-Arroyo, Olga
Ortega, Ana
Perez-Hernandez, Javier
Chaves, Felipe J.
Redon, Josep
Cortes, Raquel
author_facet Martinez-Arroyo, Olga
Ortega, Ana
Perez-Hernandez, Javier
Chaves, Felipe J.
Redon, Josep
Cortes, Raquel
author_sort Martinez-Arroyo, Olga
collection PubMed
description Kidney injury in hypertension and diabetes entails, among in other structures, damage in a key cell of the glomerular filtration barrier, the podocyte. Podocytes are polarized and highly differentiated cells in which vesicular transport, partly driven by Rab GTPases, is a relevant process. The aim of the present study was to analyze Rab GTPases of the Rab-Rabphilin system in human immortalized podocytes and the impact of high glucose and angiotensin II. Furthermore, alterations of the system in urine cell pellets from patients with hypertension and diabetes were studied. Apoptosis was analyzed in podocytes, and mRNA level quantification, Western blot analysis, and immunofluorescence were developed to quantify podocyte-specific molecules and Rab-Rabphilin components (Rab3A, Rab27A, and Rabphilin3A). Quantitative RT-PCR was performed on urinary cell pellet from patients. The results showed that differentiated cells had reduced protein levels of the Rab-rabphillin system compared with undifferentiated cells. After glucose overload and angiotensin II treatment, apoptosis was increased and podocyte-specific proteins were reduced. Rab3A and Rab27A protein levels were increased under glucose overload, and Rabphilin3A decreased. Furthermore, this system exhibited higher levels under stress conditions in a manner of angiotensin II dose and time treatment. Immunofluorescence imaging indicated different expression patterns of podocyte markers and Rab27A under treatments. Finally, Rab3A and Rab27A were increased in patient urine pellets and showed a direct relationship with albuminuria. Collectively, these results suggest that the Rab-Rabphilin system could be involved in the alterations observed in injured podocytes and that a mechanism may be activated to reduce damage through the vesicular transport enhancement directed by this system.
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spelling pubmed-74738992021-08-01 The Rab-Rabphilin system in injured human podocytes stressed by glucose overload and angiotensin II Martinez-Arroyo, Olga Ortega, Ana Perez-Hernandez, Javier Chaves, Felipe J. Redon, Josep Cortes, Raquel Am J Physiol Renal Physiol Research Article Kidney injury in hypertension and diabetes entails, among in other structures, damage in a key cell of the glomerular filtration barrier, the podocyte. Podocytes are polarized and highly differentiated cells in which vesicular transport, partly driven by Rab GTPases, is a relevant process. The aim of the present study was to analyze Rab GTPases of the Rab-Rabphilin system in human immortalized podocytes and the impact of high glucose and angiotensin II. Furthermore, alterations of the system in urine cell pellets from patients with hypertension and diabetes were studied. Apoptosis was analyzed in podocytes, and mRNA level quantification, Western blot analysis, and immunofluorescence were developed to quantify podocyte-specific molecules and Rab-Rabphilin components (Rab3A, Rab27A, and Rabphilin3A). Quantitative RT-PCR was performed on urinary cell pellet from patients. The results showed that differentiated cells had reduced protein levels of the Rab-rabphillin system compared with undifferentiated cells. After glucose overload and angiotensin II treatment, apoptosis was increased and podocyte-specific proteins were reduced. Rab3A and Rab27A protein levels were increased under glucose overload, and Rabphilin3A decreased. Furthermore, this system exhibited higher levels under stress conditions in a manner of angiotensin II dose and time treatment. Immunofluorescence imaging indicated different expression patterns of podocyte markers and Rab27A under treatments. Finally, Rab3A and Rab27A were increased in patient urine pellets and showed a direct relationship with albuminuria. Collectively, these results suggest that the Rab-Rabphilin system could be involved in the alterations observed in injured podocytes and that a mechanism may be activated to reduce damage through the vesicular transport enhancement directed by this system. American Physiological Society 2020-08-01 2020-06-22 /pmc/articles/PMC7473899/ /pubmed/32567349 http://dx.doi.org/10.1152/ajprenal.00077.2020 Text en Copyright © 2020 the American Physiological Society http://creativecommons.org/licenses/by/4.0/deed.en_US Licensed under Creative Commons Attribution CC-BY 4.0 (http://creativecommons.org/licenses/by/4.0/deed.en_US) : © the American Physiological Society.
spellingShingle Research Article
Martinez-Arroyo, Olga
Ortega, Ana
Perez-Hernandez, Javier
Chaves, Felipe J.
Redon, Josep
Cortes, Raquel
The Rab-Rabphilin system in injured human podocytes stressed by glucose overload and angiotensin II
title The Rab-Rabphilin system in injured human podocytes stressed by glucose overload and angiotensin II
title_full The Rab-Rabphilin system in injured human podocytes stressed by glucose overload and angiotensin II
title_fullStr The Rab-Rabphilin system in injured human podocytes stressed by glucose overload and angiotensin II
title_full_unstemmed The Rab-Rabphilin system in injured human podocytes stressed by glucose overload and angiotensin II
title_short The Rab-Rabphilin system in injured human podocytes stressed by glucose overload and angiotensin II
title_sort rab-rabphilin system in injured human podocytes stressed by glucose overload and angiotensin ii
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7473899/
https://www.ncbi.nlm.nih.gov/pubmed/32567349
http://dx.doi.org/10.1152/ajprenal.00077.2020
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