Cell lines

umbilical cord blood-derived endothelial progenitor cells

Preparation Method

UCB EPCs (umbilical cord blood-derived endothelial progenitor cells) were cultured in Endothelial Cell Growth Medium-2 (EGM-2) media or conditioned media (CM) from human CECs, with and without the addition of Y-27632 (10 μM).

Reaction Conditions

10 μM;

Applications

Culturing UCB EPCs in conditioned media supplemented with 10 μM Y-27632 resulted in higher proliferation rates compared with EGM-2 media and conditioned media.

Animal models

Male balb/c mice

Preparation Method

Male balb/c mice (n=8, for each group) were used in the experiment. Hot-plate latency and the number of writhes were recorded in control and in Y-27632-treated (1-5 mg/kg, i.p.) groups.

Dosage form

1-5 mg/kg, i.p.

Applications

Y-27632 (1 mg/kg) did not affect hot-plate latency; however, it considerably diminished the number of writhes, from 89+/-12 in control to 30+/-6 in the mice treated with 1 mg/kg Y-27632. At a higher dose (5 mg/kg), Y-27632 prolonged the hot-plate latency from 8.7+/-1.0 s to 14.4+/-1.7 s and decreased the number of writhes from 80+/-8 to 24+/-7.

Y-27632 dihydrochloride, as a selective Rho-kinase inhibitor, is a novel bronchodilator.^[1]

In vitro, treatment with 3 and 10 μM Y-27632 remarkably reduced the maximal contractile response. And Y-27632 (10 μM ) markedly increased the EFS-induced outflow of radioactivity from airway cholinergic nerves by 27% and 54% respectively, in murine and guinea-pig tracheal preparations loaded with [(3)H]-choline.^[2]In vitro, Y-27632 at 10 μM abolished stress fibers in Swiss 3T3 cells, but it had no effect in the G(1)-S phase transition of the cell cycle and cytokinesis.^[3]In vitro, treatment with 10 μM Y-27632 and hypoxia further increased the expression of ACAN and COL2A1 in chondrocytic cells.^[4]In vitro, 100 μM Y-27632 significantly promoted cell proliferation and phagocytosis of trabecular meshwork cells.^[6]

In vivo efficacy test it shown that Y-27632 inhalation (1 mM, 2 min) inhibited acetylcholine- or ovalbumin-induced increase in R(L) and had no changes in mean blood pressure, and the effect last for at least 3 h.^[1]In vivo experiment it indicated that injection 5mg/kg Y-27632 intravenously markedly decreased the serum levels of interleukin-6 (IL-6), IL-1β, tumor necrosis factor-α (TNF-α) and increased IL-10 level in serum of MRL/lpr mice.^[5]In addition, treatment with 2 or 30 mg/kg body weight of Y-27632 orally in SOD1(G93A) mice, Y-27632 improved motor function in male mice at 30 mg/kg, but it had no benefit at 2 mg/kg.^[7]

References:
[1]Iizuka K, et al. Evaluation of Y-27632, a rho-kinase inhibitor, as a bronchodilator in guinea pigs. Eur J Pharmacol. 2000 Oct 13;406(2):273-9.
[2]Fernandes L, et al. A Rho-kinase inhibitor, Y-27632, reduces cholinergic contraction but not neurotransmitter release. Eur J Pharmacol. 2006 Nov 21;550(1-3):155-61.
[3]Ishizaki T, et al. Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases. Mol Pharmacol. 2000 May;57(5):976-83.
[4]Piltti J, et al. Rho-kinase inhibitor Y-27632 and hypoxia synergistically enhance chondrocytic phenotype and modify S100 protein profiles in human chondrosarcoma cells. Sci Rep. 2017 Jun 16;7(1):3708.
[5]Wang Y, et al. Y-27632, a Rho-associated protein kinase inhibitor, inhibits systemic lupus erythematosus. Biomed Pharmacother. 2017 Apr;88:359-366.
[6]Chen W, et al. Rho-Associated Protein Kinase Inhibitor Treatment Promotes Proliferation and Phagocytosis in Trabecular Meshwork Cells. Front Pharmacol. 2020 Mar 17;11:302.
[7]Günther R, et al. The rho kinase inhibitor Y-27632 improves motor performance in male SOD1(G93A) mice. Front Neurosci. 2014 Oct 7;8:304.