Rosiglitasone and ROCK Inhibitors Modulate Fibrogenetic Changes in TGF-β2 Treated Human Conjunctival Fibroblasts (HconF) in Different Manners
Purpose: The effects of Rho-associated coiled-coil containing protein kinase (ROCK) 1 and 2 inhibitor, ripasudil hydrochloride hydrate (Rip), ROCK2 inhibitor, KD025 or rosiglitazone (Rosi) on two-dimension (2D) and three-dimension (3D) cultured human conjunctival fibroblasts (HconF) treated by trans...
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MDPI AG
2021-07-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/14/7335 |
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| author | Yuika Oouchi Megumi Watanabe Yosuke Ida Hiroshi Ohguro Fumihito Hikage |
| author_facet | Yuika Oouchi Megumi Watanabe Yosuke Ida Hiroshi Ohguro Fumihito Hikage |
| author_sort | Yuika Oouchi |
| collection | DOAJ |
| description | Purpose: The effects of Rho-associated coiled-coil containing protein kinase (ROCK) 1 and 2 inhibitor, ripasudil hydrochloride hydrate (Rip), ROCK2 inhibitor, KD025 or rosiglitazone (Rosi) on two-dimension (2D) and three-dimension (3D) cultured human conjunctival fibroblasts (HconF) treated by transforming growth factor (TGFβ2) were studied. Methods: Two-dimension and three-dimension cultured HconF were examined by transendothelial electrical resistance (TEER, 2D), size and stiffness (3D), and the expression of the extracellular matrix (ECM) including collagen1 (<i>COL1</i>), <i>COL4</i> and <i>COL6</i>, fibronectin <i>(FN</i>), and α-smooth muscle actin <i>(αSMA</i>) by quantitative PCR (2D, 3D) in the presence of Rip, KD025 or Rosi. Results: TGFβ2 caused a significant increase in (1) the TEER values (2D) which were greatly reduced by Rosi, (2) the stiffness of the 3D organoids which were substantially reduced by Rip or KD025, and (3) TGFβ2 induced a significant up-regulation of all ECMs, except for <i>COL6</i> (2D) or <i>αSMA</i> (3D), and down-regulation of <i>COL6</i> (2D). Rosi caused a significant up-regulation of <i>COL1, 4</i> and <i>6</i> (3D), and down-regulation of <i>COL6</i> (2D) and <i>αSMA</i> (3D). Most of these TGFβ2-induced expressions in the 2D and <i>αSMA</i> in the 3D were substantially inhibited by KD025, but <i>COL4</i> and <i>αSMA</i> in 2D were further enhanced by Rip. Conclusion: The findings reported herein indicate that TGFβ2 induces an increase in fibrogenetic changes on the plane and in the spatial space, and are inhibited by Rosi and ROCK inhibitors, respectively. |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
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| spelling | doaj.art-b2914c58a5bf449c85ace6f4f546f9e12023-11-22T03:56:54ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-012214733510.3390/ijms22147335Rosiglitasone and ROCK Inhibitors Modulate Fibrogenetic Changes in TGF-β2 Treated Human Conjunctival Fibroblasts (HconF) in Different MannersYuika Oouchi0Megumi Watanabe1Yosuke Ida2Hiroshi Ohguro3Fumihito Hikage4Departments of Ophthalmology, School of Medicine, Sapporo Medical University, S1W17, Chuo-ku, Sapporo 060-8556, JapanDepartments of Ophthalmology, School of Medicine, Sapporo Medical University, S1W17, Chuo-ku, Sapporo 060-8556, JapanDepartments of Ophthalmology, School of Medicine, Sapporo Medical University, S1W17, Chuo-ku, Sapporo 060-8556, JapanDepartments of Ophthalmology, School of Medicine, Sapporo Medical University, S1W17, Chuo-ku, Sapporo 060-8556, JapanDepartments of Ophthalmology, School of Medicine, Sapporo Medical University, S1W17, Chuo-ku, Sapporo 060-8556, JapanPurpose: The effects of Rho-associated coiled-coil containing protein kinase (ROCK) 1 and 2 inhibitor, ripasudil hydrochloride hydrate (Rip), ROCK2 inhibitor, KD025 or rosiglitazone (Rosi) on two-dimension (2D) and three-dimension (3D) cultured human conjunctival fibroblasts (HconF) treated by transforming growth factor (TGFβ2) were studied. Methods: Two-dimension and three-dimension cultured HconF were examined by transendothelial electrical resistance (TEER, 2D), size and stiffness (3D), and the expression of the extracellular matrix (ECM) including collagen1 (<i>COL1</i>), <i>COL4</i> and <i>COL6</i>, fibronectin <i>(FN</i>), and α-smooth muscle actin <i>(αSMA</i>) by quantitative PCR (2D, 3D) in the presence of Rip, KD025 or Rosi. Results: TGFβ2 caused a significant increase in (1) the TEER values (2D) which were greatly reduced by Rosi, (2) the stiffness of the 3D organoids which were substantially reduced by Rip or KD025, and (3) TGFβ2 induced a significant up-regulation of all ECMs, except for <i>COL6</i> (2D) or <i>αSMA</i> (3D), and down-regulation of <i>COL6</i> (2D). Rosi caused a significant up-regulation of <i>COL1, 4</i> and <i>6</i> (3D), and down-regulation of <i>COL6</i> (2D) and <i>αSMA</i> (3D). Most of these TGFβ2-induced expressions in the 2D and <i>αSMA</i> in the 3D were substantially inhibited by KD025, but <i>COL4</i> and <i>αSMA</i> in 2D were further enhanced by Rip. Conclusion: The findings reported herein indicate that TGFβ2 induces an increase in fibrogenetic changes on the plane and in the spatial space, and are inhibited by Rosi and ROCK inhibitors, respectively.https://www.mdpi.com/1422-0067/22/14/7335TGFβ2human conjunctival fibroblastROCK inhibitor3D cultureripasudilKD025 |
| spellingShingle | Yuika Oouchi Megumi Watanabe Yosuke Ida Hiroshi Ohguro Fumihito Hikage Rosiglitasone and ROCK Inhibitors Modulate Fibrogenetic Changes in TGF-β2 Treated Human Conjunctival Fibroblasts (HconF) in Different Manners International Journal of Molecular Sciences TGFβ2 human conjunctival fibroblast ROCK inhibitor 3D culture ripasudil KD025 |
| title | Rosiglitasone and ROCK Inhibitors Modulate Fibrogenetic Changes in TGF-β2 Treated Human Conjunctival Fibroblasts (HconF) in Different Manners |
| title_full | Rosiglitasone and ROCK Inhibitors Modulate Fibrogenetic Changes in TGF-β2 Treated Human Conjunctival Fibroblasts (HconF) in Different Manners |
| title_fullStr | Rosiglitasone and ROCK Inhibitors Modulate Fibrogenetic Changes in TGF-β2 Treated Human Conjunctival Fibroblasts (HconF) in Different Manners |
| title_full_unstemmed | Rosiglitasone and ROCK Inhibitors Modulate Fibrogenetic Changes in TGF-β2 Treated Human Conjunctival Fibroblasts (HconF) in Different Manners |
| title_short | Rosiglitasone and ROCK Inhibitors Modulate Fibrogenetic Changes in TGF-β2 Treated Human Conjunctival Fibroblasts (HconF) in Different Manners |
| title_sort | rosiglitasone and rock inhibitors modulate fibrogenetic changes in tgf β2 treated human conjunctival fibroblasts hconf in different manners |
| topic | TGFβ2 human conjunctival fibroblast ROCK inhibitor 3D culture ripasudil KD025 |
| url | https://www.mdpi.com/1422-0067/22/14/7335 |
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