Effect of Rho-associated kinase inhibitor on human corneal endothelial cell apoptosis

J Cataract Refract Surg. 2020 Apr;46(4):612-616. doi: 10.1097/j.jcrs.0000000000000115.

Abstract

Purpose: To evaluate whether exposure to Rho-associated protein kinase (ROCK) inhibitor will promote human-cultured corneal endothelial cells (CECs) survival in a commercial storage medium.

Setting: Edith Wolfson Medical Center, Holon, and Sheba Medical Center, Tel Hashomer, Israel.

Design: Experimental study.

Methods: Fragments of human donor corneolimbal rings were stored in commercial storage media for 1 week, half with the addition of 10 μM ROCK inhibitor (Y-27632). Evaluation of CECs for early and late apoptosis\necrosis rates was performed using anti-human CD166 antibody and flow cytometric double staining analysis of propidium iodide and Annexin V.

Results: CECs of 6 corneolimbal rings demonstrated a reduced early apoptosis rate (4.35% ± 1.07% vs 12.18% ± 5.5%, P = .026) and a reduced late apoptosis\necrosis rate (5.5% ± 2.39% vs 9.43% ± 2.61%, P = .004) compared with control. Subsequently, the rate of apoptotic CECs expressing ROCK was significantly lower in cells exposed to ROCK inhibitor compared with cells that were not (19.01% ± 4.17 vs 30.42% ± 4.27, P < .001).

Conclusions: ROCK inhibitor reduced endothelial cell loss in vitro and might be used to limit or slow CEC loss in donor corneal tissue during eye banking. This might be a promising new method for promoting future graft survival.

MeSH terms

  • Adult
  • Aged
  • Amides / pharmacology*
  • Annexin A5 / metabolism
  • Antigens, CD / metabolism
  • Apoptosis / drug effects*
  • Cell Adhesion Molecules, Neuronal / metabolism
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Culture Media
  • Endothelium, Corneal / drug effects*
  • Endothelium, Corneal / metabolism
  • Endothelium, Corneal / pathology
  • Enzyme Inhibitors / pharmacology*
  • Female
  • Fetal Proteins / metabolism
  • Flow Cytometry
  • Humans
  • Male
  • Middle Aged
  • Pyridines / pharmacology*
  • Tissue Donors
  • rho-Associated Kinases / antagonists & inhibitors*

Substances

  • ALCAM protein, human
  • Amides
  • Annexin A5
  • Antigens, CD
  • Cell Adhesion Molecules, Neuronal
  • Culture Media
  • Enzyme Inhibitors
  • Fetal Proteins
  • Pyridines
  • Y 27632
  • rho-Associated Kinases