May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Ache Expression in Y79 Cells During Apoptosis Resulting From Hyperglycemia
Author Affiliations & Notes
  • R. S. Shehadeh Masha'our
    Physiology- Eye Research Unit, Faculty of Medicine- Technion, Haifa, Israel
  • R. Heinrich
    Physiology- Eye Research Unit, Faculty of Medicine- Technion, Haifa, Israel
  • H. J. Garzozi
    Department of Ophthalmology, Bnai-Zion Medical Center, Haifa, Israel
  • I. Perlman
    Physiology- Eye Research Unit, Faculty of Medicine- Technion, Haifa, Israel
  • Footnotes
    Commercial Relationships  R.S. Shehadeh Masha'our, None; R. Heinrich, None; H.J. Garzozi, None; I. Perlman, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 4900. doi:
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      R. S. Shehadeh Masha'our, R. Heinrich, H. J. Garzozi, I. Perlman; Ache Expression in Y79 Cells During Apoptosis Resulting From Hyperglycemia. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4900.

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

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Abstract

Purpose: : : Increasing evidence has shown that AChE may be involved in stress-induced retinal cells’ apoptosis. Here, we tested the effects of hyperglycemia, as a form of metabolic stress, on retinal cells in culture, and on AChE expression and its relation to apoptosis.

Methods: : Y79 cells were cultured for 24 hours in starvation medium containing 0.1% glucose, mimicking physiologic range of blood glucose concentration, and 1% FBS. Then, the cells were transferred for 1-24 hours to a medium containing high glucose concentration, range 0.2%-0.6%. To control for hyperosmolarity, the effects of similar concentrations of mannitol were tested. Apoptosis was determined by PARP cleavage and TUNEL staining. AChE protein expression was determined by Western blot and activity by Karnovsky method.

Results: : PARP cleavage was observed in cells treated with the addition of 0.25% and 0.5% glucose for 1 and 2 hours, mimicking glucose concentrations in diabetes, as compared to 0.1% glucose, and in cells treated with similar concentrations of mannitol. PARP cleavage was similar in cells treated with 0.5% glucose or mannitol, but was more pronounced in cells treated with 0.25% glucose compared to 0.25% mannitol. Treatment in 0.25% and 0.5% glucose for 16 and 24 hours increased significantly TUNEL positive cells compared to cells exposed to 0.1% glucose. AChE expression was increased in cells treated with the addition of 0.25% glucose for an hour compared to control, but not in cells treated with higher concentrations or longer incubation, or in those treated with mannitol. AChE activity was increased significantly in cells treated with the addition of 0.25% glucose for 1 hour as compared to 0.1% and to a lesser degree in cells treated with additional 0.25% for 16 and 24 hours.

Conclusions: : Hyperglycemia induces apoptosis in retinal cells line in a process that may involve AChE. As hyperglycemia is a major pathogenic factor in the development of diabetic retinopathy, understanding the mechanisms of hyperglycemia induced apoptosis is crucial for understanding and treating diabetic retinopathy.

Keywords: acetylcholine • apoptosis/cell death • diabetic retinopathy 
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