April 2011
Volume 52, Issue 14
ARVO Annual Meeting Abstract  |   April 2011
Analysis of Oxidative Stress in Diabetic Rat Retina and in Cultured Retinal Cells Under Hyperglycemic Conditions
Author Affiliations & Notes
  • Mohammad S. Ola
    King Saud University-Ophthal, King Abdul Aziz Univ Hosp-Coll of Med, Riyadh, Saudi Arabia
  • Kathryn F. LaNoue
    Cellular & Molecular Physiology, Penn State College of Medicine, Hershey, Pennsylvania
  • Footnotes
    Commercial Relationships  Mohammad S. Ola, None; Kathryn F. LaNoue, None
  • Footnotes
    Support  Dr.Nasir Al-Rasheed Research Chair in Ophthalmology and JDRF
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 4450. doi:
  • Views
  • Share
  • Tools
    • Alerts
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Mohammad S. Ola, Kathryn F. LaNoue; Analysis of Oxidative Stress in Diabetic Rat Retina and in Cultured Retinal Cells Under Hyperglycemic Conditions. Invest. Ophthalmol. Vis. Sci. 2011;52(14):4450.

      Download citation file:

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

  • Supplements

Purpose: : Numerous studies show that hyperglycemia leads to oxidative stress in the diabetic retinas, but the mechanisms that generate oxidative stress have not been resolved. Increased electron pressure on the mitochondrial electron transfer chain, increased generation of cytosolic NADH, and decreases in cellular NADPH have all been cited as possible sources of reactive oxygen species (ROS) and nitric oxide. The purpose of this study was to determine the level of oxidative stress in the ex vivo diabetic retina and in retinal endothelial and Muller cells under hyperglycemic conditions.

Methods: : Catalase activity and the level of H2O2 were determined in ex vivo control and 2 months streptozotocin-induced diabetic retina using Fluro H2O2TM kit. Cultured bovine retinal endothelial cells and Muller cells (TR-MUL) were incubated either with 5 or 30 mM glucose for 1 to 5 days and generation of ROS measured as oxidized fluorescent product of CM-H2DCFDA. ROS was also measured in endothelial cells treated with pyruvate (5mM) and glutamine (5mM). TR-MUL cells were also treated with diamide (200 µM), CuSO4 (20 µM), pyruvate (5 mM) and glutamine (5-10 mM) in presence and absence of high glucose and the level of ROS measured. Diamide and CuSO4 cause oxidative stress in cells.

Results: : Catalase activities were found to be the similar in both control and diabetic retina. An increased level of H2O2 was detected in the diabetic retina compared to non-diabetic when catalase activity was inhibited with CuSO4. In contradiction to previous studies by others, our study demonstrated that high glucose exposure to both endothelial and Muller cells for 1 to 5 days, the level of ROS significantly reduced (P < 0.05). Treatments with pyruvate could not influence the increase in ROS generation in these cells, however, glutamine treatments caused a significant increase in ROS generation in both endothelial and Muller cells but more dramatically in endothelial cells.

Conclusions: : High levels of glucose may not provide the mitochondria with excess reducing pressure that would generate superoxide anions by reduction of molecular oxygen at intermediate steps in the electron transport chain. However, diabetes influences an increase in oxidative stress in the retina.

Keywords: oxidation/oxidative or free radical damage • diabetic retinopathy • retina 

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.