ROCK Inhibitor Y-27632 Promotes Human Retinal Pigment Epithelium Survival by Altering Cellular Biomechanical Properties

Author(s): Y. Ni, Y. Qin, Z. Fang, Z. Zhang*.

Journal Name: Current Molecular Medicine

Volume 17 , Issue 9 , 2017

  Journal Home
Translate in Chinese
Submit Manuscript
Submit Proposal

Abstract:

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 degenerative diseases.

Keywords: Retinal pigment epithelial cells, ROCK inhibitor, Y-27632, proliferation, cytoskeleton, mechanical properties.

Rights & PermissionsPrintExport Cite as


Article Details

VOLUME: 17
ISSUE: 9
Year: 2017
Page: [637 - 646]
Pages: 10
DOI: 10.2174/1566524018666180316150936

Article Metrics

PDF: 51
HTML: 8