Cargando…
An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model
We report herein on the effects of brimonidine (BRI), an α2-adrenergic agonist, on two-dimensional (2D) and three-dimensional (3D) cell-cultured TGF-β2-untreated and -treated human trabecular meshwork (HTM) cells. In the presence of TGF-β2 (5 ng/mL), (1) the effects of BRI on (1) the 2D HTM monolaye...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9312232/ https://www.ncbi.nlm.nih.gov/pubmed/35877361 http://dx.doi.org/10.3390/bioengineering9070310 |
_version_ | 1784753794188312576 |
---|---|
author | Watanabe, Megumi Sato, Tatsuya Tsugeno, Yuri Higashide, Megumi Furuhashi, Masato Umetsu, Araya Suzuki, Soma Ida, Yosuke Hikage, Fumihito Ohguro, Hiroshi |
author_facet | Watanabe, Megumi Sato, Tatsuya Tsugeno, Yuri Higashide, Megumi Furuhashi, Masato Umetsu, Araya Suzuki, Soma Ida, Yosuke Hikage, Fumihito Ohguro, Hiroshi |
author_sort | Watanabe, Megumi |
collection | PubMed |
description | We report herein on the effects of brimonidine (BRI), an α2-adrenergic agonist, on two-dimensional (2D) and three-dimensional (3D) cell-cultured TGF-β2-untreated and -treated human trabecular meshwork (HTM) cells. In the presence of TGF-β2 (5 ng/mL), (1) the effects of BRI on (1) the 2D HTM monolayers’ barrier function were investigated as estimated using trans-endothelial electrical resistance (TEER) measurement and FITC dextran permeability; (2) real-time analyses of cellular metabolism using a Seahorse Bioanalyzer; (3) the largeness and hardness of 3D spheroids; and (4) the expression of genes that encode extracellular matrix (ECM) proteins, including collagens (COL) 1, 4, and 6; fibronectin (FN) and α-smooth muscle actin (α-SMA); ECM modulators, including a tissue inhibitor of matrix proteinase (TIMP) 1–4; matrix metalloproteinase (MMP) 2, 9, and 14; and several endoplasmic reticulum (ER) stress-related genes, including the X-box-binding protein 1 (XBP1), the spliced XBP1 (sXBP1), glucose-regulated protein (GRP)78, GRP94, and CCAAT-enhancer-binding protein homologous protein (CHOP). BRI markedly inhibited the TGF-β2-induced increase in the values of TEER of the 2D cell monolayer and the hardness of the 3D spheroids, although it had no effect on their sizes. BRI also cancelled the TGF-β2-induced reduction in mitochondrial maximal respiration but had no effect on the glycolytic capacity. In addition, the gene expression of these molecules was quite different between the 2D and 3D cultures of HTM cells. The present observations found in this study indicate that BRI may beneficially affect TGF-β2-induced changes in both cultures, 2D and 3D, of HTM cells, although their structural and functional properties that were altered varied significantly between both cultures of HTM cells. |
format | Online Article Text |
id | pubmed-9312232 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-93122322022-07-26 An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model Watanabe, Megumi Sato, Tatsuya Tsugeno, Yuri Higashide, Megumi Furuhashi, Masato Umetsu, Araya Suzuki, Soma Ida, Yosuke Hikage, Fumihito Ohguro, Hiroshi Bioengineering (Basel) Article We report herein on the effects of brimonidine (BRI), an α2-adrenergic agonist, on two-dimensional (2D) and three-dimensional (3D) cell-cultured TGF-β2-untreated and -treated human trabecular meshwork (HTM) cells. In the presence of TGF-β2 (5 ng/mL), (1) the effects of BRI on (1) the 2D HTM monolayers’ barrier function were investigated as estimated using trans-endothelial electrical resistance (TEER) measurement and FITC dextran permeability; (2) real-time analyses of cellular metabolism using a Seahorse Bioanalyzer; (3) the largeness and hardness of 3D spheroids; and (4) the expression of genes that encode extracellular matrix (ECM) proteins, including collagens (COL) 1, 4, and 6; fibronectin (FN) and α-smooth muscle actin (α-SMA); ECM modulators, including a tissue inhibitor of matrix proteinase (TIMP) 1–4; matrix metalloproteinase (MMP) 2, 9, and 14; and several endoplasmic reticulum (ER) stress-related genes, including the X-box-binding protein 1 (XBP1), the spliced XBP1 (sXBP1), glucose-regulated protein (GRP)78, GRP94, and CCAAT-enhancer-binding protein homologous protein (CHOP). BRI markedly inhibited the TGF-β2-induced increase in the values of TEER of the 2D cell monolayer and the hardness of the 3D spheroids, although it had no effect on their sizes. BRI also cancelled the TGF-β2-induced reduction in mitochondrial maximal respiration but had no effect on the glycolytic capacity. In addition, the gene expression of these molecules was quite different between the 2D and 3D cultures of HTM cells. The present observations found in this study indicate that BRI may beneficially affect TGF-β2-induced changes in both cultures, 2D and 3D, of HTM cells, although their structural and functional properties that were altered varied significantly between both cultures of HTM cells. MDPI 2022-07-12 /pmc/articles/PMC9312232/ /pubmed/35877361 http://dx.doi.org/10.3390/bioengineering9070310 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 Watanabe, Megumi Sato, Tatsuya Tsugeno, Yuri Higashide, Megumi Furuhashi, Masato Umetsu, Araya Suzuki, Soma Ida, Yosuke Hikage, Fumihito Ohguro, Hiroshi An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model |
title | An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model |
title_full | An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model |
title_fullStr | An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model |
title_full_unstemmed | An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model |
title_short | An α2-Adrenergic Agonist, Brimonidine, Beneficially Affects the TGF-β2-Treated Cellular Properties in an In Vitro Culture Model |
title_sort | α2-adrenergic agonist, brimonidine, beneficially affects the tgf-β2-treated cellular properties in an in vitro culture model |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9312232/ https://www.ncbi.nlm.nih.gov/pubmed/35877361 http://dx.doi.org/10.3390/bioengineering9070310 |
work_keys_str_mv | AT watanabemegumi ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT satotatsuya ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT tsugenoyuri ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT higashidemegumi ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT furuhashimasato ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT umetsuaraya ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT suzukisoma ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT idayosuke ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT hikagefumihito ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT ohgurohiroshi ana2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT watanabemegumi a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT satotatsuya a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT tsugenoyuri a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT higashidemegumi a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT furuhashimasato a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT umetsuaraya a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT suzukisoma a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT idayosuke a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT hikagefumihito a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel AT ohgurohiroshi a2adrenergicagonistbrimonidinebeneficiallyaffectsthetgfb2treatedcellularpropertiesinaninvitroculturemodel |