September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Cyclosporine A-induced Senescence in Cultured Human Corneal Endothelial Cells
Author Affiliations & Notes
  • Ki Won Jin
    Department of Ophthalmology, Hallym University College of Medicine, Seoul, Korea (the Republic of)
  • Sang Wook Choi
    Department of Ophthalmology, Hallym University College of Medicine, Seoul, Korea (the Republic of)
  • Joon-Young Hyon
    Department of Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Korea (the Republic of)
    Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea (the Republic of)
  • Tae-Young Chung
    Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea (the Republic of)
  • Won Ryang Wee
    Department of Ophthalmology, Seoul National University College of Medicine, Seoul, Korea (the Republic of)
  • Young Joo Shin
    Department of Ophthalmology, Hallym University College of Medicine, Seoul, Korea (the Republic of)
  • Footnotes
    Commercial Relationships   Ki Won Jin, None; Sang Wook Choi, None; Joon-Young Hyon, None; Tae-Young Chung, None; Won Ryang Wee, None; Young Joo Shin, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 5297. doi:
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      Ki Won Jin, Sang Wook Choi, Joon-Young Hyon, Tae-Young Chung, Won Ryang Wee, Young Joo Shin; Cyclosporine A-induced Senescence in Cultured Human Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5297.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : To investigate the effect of cyclosporine A (CsA) on senescence in human corneal endothelial cells (HCECs).

Methods : HCECs were cultured and treated with a various concentration of CsA (0-100µM). Senescence-associated β–galactosidase (SA-β-gal staining) staining was performed. Mitochondrial dehydrogenase activity was assessed using a WST-8 assay kit and mitochondrial membrane potential (ΔΨm) was measured using JC-1. Intracellular and mitochondrial reactive oxygen species formation was measured by DCF-DA and MitoSOX probe. Intracellular calcium levels were measured using Fluo-4, and mitochondrial calcium levels were measured using Rhod-2. Protein expression of glycogen synthase kinase 3 beta (GSK3β), pGSK3β, ERK 1/2 (extracellular-signal-regulated kinases 1/2) and pERK 1/2 was evaluated by western blotting.

Results : CsA increased Percentage of senescence associated (SA)-gal positive cells increased in a dose-dependent manner (p<0.05). CsA decreased mitochondrial dehydrogenase activity and ΔΨm in a dose-dependent manner (p<0.05). Intracellular and mitochondrial reactive oxygen species levels increased in treatment with CsA (p<0.05). CsA increased mitochondrial calcium levels at 100 µM (p<0.05) whereas intracellular calcium levels decreased at 100 µM. CsA attenuated the phosphorylation of GSK3β and ERK1/2.

Conclusions : CsA induced senescence in HCECs through induction of oxidative stress and via mitochodnrial/GSK3β/ERK1/2 dependent pathways.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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