Cargando…

The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner

The purpose of the present study was to examine the effect of the selective α1 antagonist tamsulosin (TAM) on human retinal pigment epithelium cells, ARPE 19. Two-dimension (2D) and three-dimension (3D) cultured ARPE 19 cells were used in the following characterizations: (1) ultrastructure by scanni...

Descripción completa

Detalles Bibliográficos
Autores principales: Ida, Yosuke, Sato, Tatsuya, Watanabe, Megumi, Umetsu, Araya, Tsugeno, Yuri, Furuhashi, Masato, Hikage, Fumihito, Ohguro, Hiroshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9598783/
https://www.ncbi.nlm.nih.gov/pubmed/36290524
http://dx.doi.org/10.3390/bioengineering9100556
_version_ 1784816436940636160
author Ida, Yosuke
Sato, Tatsuya
Watanabe, Megumi
Umetsu, Araya
Tsugeno, Yuri
Furuhashi, Masato
Hikage, Fumihito
Ohguro, Hiroshi
author_facet Ida, Yosuke
Sato, Tatsuya
Watanabe, Megumi
Umetsu, Araya
Tsugeno, Yuri
Furuhashi, Masato
Hikage, Fumihito
Ohguro, Hiroshi
author_sort Ida, Yosuke
collection PubMed
description The purpose of the present study was to examine the effect of the selective α1 antagonist tamsulosin (TAM) on human retinal pigment epithelium cells, ARPE 19. Two-dimension (2D) and three-dimension (3D) cultured ARPE 19 cells were used in the following characterizations: (1) ultrastructure by scanning electron microscopy (SEM) (2D); (2) barrier functions by transepithelial electrical resistance (TEER) measurements, and FITC-dextran permeability (2D); (3) real time cellular metabolisms by Seahorse Bioanalyzer (2D); (4) physical properties, size and stiffness measurements (3D); and (5) expression of extracellular matrix (ECM) proteins, including collagen1 (COL1), COL4, COL6 and fibronectin (FN) by qPCR and immunohistochemistry (2D and 3D). TAM induced significant effects including: (1) alteration of the localization of the ECM deposits; (2) increase and decrease of the TEER values and FITC-dextran permeability, respectively; (3) energy shift from glycolysis into mitochondrial oxidative phosphorylation (OXPHOS); (4) large and stiffened 3D spheroids; and (5) down-regulations of the mRNA expressions and immune labeling of most ECM proteins in a concentration-dependent manner. However, in some ECM proteins, COL1 and COL6, their immunolabeling intensities were increased at the lowest concentration (1 μM) of TAM. Such a discrepancy between the gene expressions and immunolabeling of ECM proteins may support alterations of ECM localizations as observed by SEM. The findings reported herein indicate that the selective α1 antagonist, TAM, significantly influenced ECM production and distribution as well as cellular metabolism levels in a concentration-dependent manner.
format Online
Article
Text
id pubmed-9598783
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-95987832022-10-27 The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner Ida, Yosuke Sato, Tatsuya Watanabe, Megumi Umetsu, Araya Tsugeno, Yuri Furuhashi, Masato Hikage, Fumihito Ohguro, Hiroshi Bioengineering (Basel) Article The purpose of the present study was to examine the effect of the selective α1 antagonist tamsulosin (TAM) on human retinal pigment epithelium cells, ARPE 19. Two-dimension (2D) and three-dimension (3D) cultured ARPE 19 cells were used in the following characterizations: (1) ultrastructure by scanning electron microscopy (SEM) (2D); (2) barrier functions by transepithelial electrical resistance (TEER) measurements, and FITC-dextran permeability (2D); (3) real time cellular metabolisms by Seahorse Bioanalyzer (2D); (4) physical properties, size and stiffness measurements (3D); and (5) expression of extracellular matrix (ECM) proteins, including collagen1 (COL1), COL4, COL6 and fibronectin (FN) by qPCR and immunohistochemistry (2D and 3D). TAM induced significant effects including: (1) alteration of the localization of the ECM deposits; (2) increase and decrease of the TEER values and FITC-dextran permeability, respectively; (3) energy shift from glycolysis into mitochondrial oxidative phosphorylation (OXPHOS); (4) large and stiffened 3D spheroids; and (5) down-regulations of the mRNA expressions and immune labeling of most ECM proteins in a concentration-dependent manner. However, in some ECM proteins, COL1 and COL6, their immunolabeling intensities were increased at the lowest concentration (1 μM) of TAM. Such a discrepancy between the gene expressions and immunolabeling of ECM proteins may support alterations of ECM localizations as observed by SEM. The findings reported herein indicate that the selective α1 antagonist, TAM, significantly influenced ECM production and distribution as well as cellular metabolism levels in a concentration-dependent manner. MDPI 2022-10-14 /pmc/articles/PMC9598783/ /pubmed/36290524 http://dx.doi.org/10.3390/bioengineering9100556 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ida, Yosuke
Sato, Tatsuya
Watanabe, Megumi
Umetsu, Araya
Tsugeno, Yuri
Furuhashi, Masato
Hikage, Fumihito
Ohguro, Hiroshi
The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner
title The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner
title_full The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner
title_fullStr The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner
title_full_unstemmed The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner
title_short The Selective α1 Antagonist Tamsulosin Alters ECM Distributions and Cellular Metabolic Functions of ARPE 19 Cells in a Concentration-Dependent Manner
title_sort selective α1 antagonist tamsulosin alters ecm distributions and cellular metabolic functions of arpe 19 cells in a concentration-dependent manner
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9598783/
https://www.ncbi.nlm.nih.gov/pubmed/36290524
http://dx.doi.org/10.3390/bioengineering9100556
work_keys_str_mv AT idayosuke theselectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT satotatsuya theselectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT watanabemegumi theselectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT umetsuaraya theselectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT tsugenoyuri theselectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT furuhashimasato theselectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT hikagefumihito theselectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT ohgurohiroshi theselectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT idayosuke selectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT satotatsuya selectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT watanabemegumi selectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT umetsuaraya selectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT tsugenoyuri selectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT furuhashimasato selectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT hikagefumihito selectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner
AT ohgurohiroshi selectivea1antagonisttamsulosinaltersecmdistributionsandcellularmetabolicfunctionsofarpe19cellsinaconcentrationdependentmanner