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 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 (), and , fibronectin ), and α-smooth muscle actin ) 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 (2D) or (3D), and down-regulation of (2D). Rosi caused a significant up-regulation of and (3D), and down-regulation of (2D) and (3D). Most of these TGFβ2-induced expressions in the 2D and in the 3D were substantially inhibited by KD025, but and 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.