High-dose glucose induced apoptosis in human retinal vascular endothelial cells

H. C. Hatcher, M. B. Grant, S. Caballero, R. K. Crouch, J. X. Ma

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Abstract

Purpose: The purpose of this study is to determine the effect of high-dose glucose on primary cultured human retinal capillary endothelial cells and its potential involvement in retinal diseases such as diabetic retinopathy. Methods: Human retinal capillary endothelial cells were obtained from donor eyes. These cells were cultured in 5mM or 25mM glucose for 6 days prior to analysis. Transmission electron microscopy (TEM) was performed on the retinal endothelial cells to observe ultrastructural events in apoptosis. Flow cytometry using the propidium iodide exclusion assay was used to quantify apoptotic cells. DNA fragmentation was analyzed by electrophoresis in an agarose gel. Results: TEM revealed extensive cell surface blebbing with preservation of the cell membrane. Cell count revealed a significant decrease in viable cells (p<0.004) when comparing cells treated with 5mM and 25mM glucose. Flow cytometry has identified a sub-G1 peak in the high-dose glucose-treated cells which indicates the existence of a large number of apoptotic cells. Apoptosis was also supported by DNA fragmentation. Conclusions: High-dose glucose (25mM) induced cell death in human retinal capillary endothelial cells through the apoptosis pathway.

Original languageEnglish (US)
Pages (from-to)S797
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996

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ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Hatcher, H. C., Grant, M. B., Caballero, S., Crouch, R. K., & Ma, J. X. (1996). High-dose glucose induced apoptosis in human retinal vascular endothelial cells. Investigative Ophthalmology and Visual Science, 37(3), S797.