April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Vascular Endothelial Growth Factor Protects Against Oxidative Stress in Differentiated Retinal Ganglion Cells
Author Affiliations & Notes
  • V. S. Brar
    Ophthalmology, University of Florida, Jacksonville, Florida
  • R. K. Sharma
    Ophthalmology, University of Florida, Jacksonville, Florida
  • R. Keshavamurthy
    Ophthalmology, University of Florida, Jacksonville, Florida
  • K. V. Chalam
    Ophthalmology, University of Florida, Jacksonville, Florida
  • Footnotes
    Commercial Relationships  V.S. Brar, None; R.K. Sharma, None; R. Keshavamurthy, None; K.V. Chalam, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 379. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      V. S. Brar, R. K. Sharma, R. Keshavamurthy, K. V. Chalam; Vascular Endothelial Growth Factor Protects Against Oxidative Stress in Differentiated Retinal Ganglion Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):379. doi: https://doi.org/.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : This experiment is aimed at evaluating potential VEGF protection against oxidative stress in a differentiated model of retinal ganglion cells and whether treatment with bevacizumab can abrogate this effect. Vascular endothelial growth factor (VEGF) has a neuroprotective function in the central nervous system neurons in conditions of hypoxia and oxidative stress. Anti-VEGF therapy is an established treatment in retinal neovascular disorders and exudative age related macular degeneration (ARMD). Oxidative stress has been implicated in the pathogenesis of ARMD. Thus, to gain a better understanding whether anti-VEGF treatment might have deleterious consequence on retinal neurons, we investigated the effect of VEGF on oxidative stress induced injury to a model of differentiated retinal ganglion cells, RGC-5.

Methods: : Rat retinal ganglion cells (RGC-5) were differentiated using the tyrosine kinase inhibitor staurosporine. Evidence of differentiation was obtained through evaluation of changes in morphology and expression of the neuronal marker class III β-tubulin by immunocytochemistry. Differentiated RGC-5 cells were treated with H2O2 for 24 hrs in the presence of increasing doses of VEGF. Cytotoxicity was assessed by quantifying cell numbers using the WST-1 assay. Specificity of oxidative stress was assessed by conducting the experiments in the presence of the glutathione reductase inhibitor BSO. Cytotoxic effects of bevacizumab were assessed by incubating differentiated RGC-5 with increasing concentrations of bevacizumab for 24h and assessing cell numbers using WST-1 and CV assays. Effect of increasing concentrations of bevacizumab on the protective effect of VEGF on H2O2 induced cell death was also assessed.

Results: : 24h treatment with staurosporine produced cell processes which were increased in number and elongated compared with untreated RGC-5. These processes were shown to be neuronal in nature through the expression of class III β-tubulin by immunocytochemistry. VEGF (200 ng/mL) protected differentiated RGC-5 against oxidative stress produced by H2O2 (800 µM). This effect was eliminated by the addition of BSO and was blocked by co-treatment with bevacizumab (2.0 mg/mL). Bevacizumab was not found to be cytotoxic at all doses tested (0.1, 1.0, & 2.0 mg/mL).

Conclusions: : VEGF protects retinal ganglion cells against oxidative stress induced cytotoxicity. This neuroprotection may rely on the glutathione reductase pathway and is blocked by bevacizumab, in a model of differentiated retinal ganglion cells.

Keywords: drug toxicity/drug effects • ganglion cells • oxidation/oxidative or free radical damage 
×
×

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.

×