Purpose: Dysfunction or death of retinal pigment epithelial (RPE) cells is a
common pathogenesis of various types of retinal degenerative diseases. Recent reports
indicated that ROCK pathway inhibitors regulate cell proliferation or apoptosis in a celltype-
dependent manner. Here, we aim to investigate the effect of ROCK inhibitor Y-
27632 on the human retinal pigment epithelium (RPE) in vitro.
Methods: Cell proliferation and apoptosis were analyzed by CCK-8 and flow cytometry
respectively. Cell proliferation markers were detected by immunofluorescence and
western blot. Cell morphology was evaluated using scanning electron microscopy. The
topography and biomechanical properties of living cells were assessed using atomic
force microscope (AFM). In addition, cytoskeleton and epithelial-mesenchymal transition
(EMT) markers were detected by western blot and immunofluorescence.
Results: 30µM Y-27632 significantly promoted cell proliferation and decreased
apoptosis. Compared with control group, human retinal pigment epithelial cell line
ARPE-19 cells treated with 30µM Y-27632 exhibited significantly decreased
cytomembrane roughness (Ra: 41.04±1.63nm vs. 24.41±0.75nm, P<0.01; Rq:
51.56±2.03nm vs. 30.81±0.95nm, P<0.01) and increased elasticity modulus
(16.66±0.83KPa vs. 32.55±1.48KPa, P<0.01). In addition, the inhibition of ROCK activity
by Y-27632 caused cell elongation and reorganization of microfilaments and
microtubules of cytoskeletons.
Conclusion: Taken together, our data demonstrated that Y-27632 could alter
biomechanical properties and reorganized cytoskeletons to promote RPE cell survival.
These results are an important step toward the future application of Y-27632 in retinal