May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Endogenous Oxidoreductase Expression is Induced by Aminoglycosides
Author Affiliations & Notes
  • J.E. Sears
    Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, OH, United States
  • Y. Chai
    Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, OH, United States
  • G. Hoppe
    Cole Eye Institute, Cleveland Clinic Foundation, Cleveland, OH, United States
  • Footnotes
    Commercial Relationships  J.E. Sears, None; Y. Chai, None; G. Hoppe, None.
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 1628. doi:
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      J.E. Sears, Y. Chai, G. Hoppe; Endogenous Oxidoreductase Expression is Induced by Aminoglycosides . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1628.

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

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Abstract

Abstract: : Purpose: To determine the effect of aminoglycosides on protein glutaredoxin-1 (grx-1) levels in cultured human retinal pigment epithelial cells. Methods: Varied concentrations of gentamycin, kanamycin, and hygromycin were used to treat ARPE-19 cells. Western blot was used to detect grx-1 content. Reactive oxygen species (ROS) formation was measured by a dichlorofluorescein diacetate (DCFDA) fluorescent assay after aminoglycoside exposure. The effect of phorbol 12-myristate 13-acetate (PMA), N-acetyl cysteine (NAC), and t-butyl hydroperoxide (tBHP) were tested in order to correlate oxidoreductase expression to ROS production. Results: All aminoglycosides increased grx-1 expression in a time and dose dependent fashion. Hygromycin was most effective, creating a 10-fold increase in grx-1 expression at a concentration of 400 µg/ml at 48 hours. PMA similarly increased grx-1 expression. NAC decreased grx-1 expression. ROS formation was correlated directly to aminoglycoside effect. Conclusions: In summary, aminoglycosides increase grx-1 protein levels. This effect is blunted by NAC, enhanced by PMA, and may be directly correlated to reactive oxygen species induced by tBHP. The effect of aminoglycoside selection on strategies for grx-1 investigation should take this observation into account. Further studies are warranted to understand the mechanism of reactive oxygen species induction of grx-1 protein levels.

Keywords: oxidation/oxidative or free radical damage • retinal pigment epithelium • cell death/apoptosis 
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