期刊
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷 22, 期 14, 页码 -出版社
MDPI
DOI: 10.3390/ijms22147335
关键词
TGF beta 2; human conjunctival fibroblast; ROCK inhibitor; 3D culture; ripasudil; KD025; rosiglitazone
The study revealed that TGF beta 2 induced fibrogenetic changes in both 2D and 3D cultured HconF, which were inhibited by Rosi and ROCK inhibitors, respectively.
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 (TGF beta 2) 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 (COL1), COL4 and COL6, fibronectin (FN), and alpha-smooth muscle actin (alpha SMA) by quantitative PCR (2D, 3D) in the presence of Rip, KD025 or Rosi. Results: TGF beta 2 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) TGF beta 2 induced a significant up-regulation of all ECMs, except for COL6 (2D) or alpha SMA (3D), and down-regulation of COL6 (2D). Rosi caused a significant up-regulation of COL1, 4 and 6 (3D), and down-regulation of COL6 (2D) and alpha SMA (3D). Most of these TGF beta 2-induced expressions in the 2D and alpha SMA in the 3D were substantially inhibited by KD025, but COL4 and alpha SMA in 2D were further enhanced by Rip. Conclusion: The findings reported herein indicate that TGF beta 2 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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