June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
Reduced apoptosis and autophagy in an oxidative stress retina organ culture model through an iNOS-inhibitor
Author Affiliations & Notes
  • Stephanie C Joachim
    Experimental Eye Research Institute, Ruhr-University Bochum, Bochum, Germany
  • Ana Maria Müller-Bühl
    Experimental Eye Research Institute, Ruhr-University Bochum, Bochum, Germany
  • Jose Hurst
    University Eye Hospital Tuebingen, Centre for Ophthalmology Tuebingen, Tuebingen, Germany
  • Carsten Theiss
    Department of Cytology, Institute of Anatomy, Ruhr-University Bochum, Bochum, Germany
  • Lisa Hofmann
    Experimental Eye Research Institute, Ruhr-University Bochum, Bochum, Germany
  • Laura Peters
    Experimental Eye Research Institute, Ruhr-University Bochum, Bochum, Germany
  • Fenja Herms
    Experimental Eye Research Institute, Ruhr-University Bochum, Bochum, Germany
  • Sandra Kuehn
    Experimental Eye Research Institute, Ruhr-University Bochum, Bochum, Germany
  • H Burkhard Dick
    Experimental Eye Research Institute, Ruhr-University Bochum, Bochum, Germany
  • Sven Schnichels
    University Eye Hospital Tuebingen, Centre for Ophthalmology Tuebingen, Tuebingen, Germany
  • Footnotes
    Commercial Relationships   Stephanie Joachim, None; Ana Maria Müller-Bühl, None; Jose Hurst, None; Carsten Theiss, None; Lisa Hofmann, None; Laura Peters, None; Fenja Herms, None; Sandra Kuehn, None; H Dick, None; Sven Schnichels, None
  • Footnotes
    Support  set Stiftung, Germany
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 2473. doi:
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      Stephanie C Joachim, Ana Maria Müller-Bühl, Jose Hurst, Carsten Theiss, Lisa Hofmann, Laura Peters, Fenja Herms, Sandra Kuehn, H Burkhard Dick, Sven Schnichels; Reduced apoptosis and autophagy in an oxidative stress retina organ culture model through an iNOS-inhibitor. Invest. Ophthalmol. Vis. Sci. 2020;61(7):2473.

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

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Abstract

Purpose : The overproduction of reactive oxygen species is defined as oxidative stress. This pathomechanism occurs in several retinal diseases, including glaucoma or retinal ischemia. Hydrogen peroxide (H2O2) can be used in vitro to simulate oxidative stress, causing a strong neurodegeneration of the inner retinal layers. Based on this model, therapeutic approaches can be tested. Hence, treatment with the nitric oxide synthase (iNOS)-inhibitor 1400W as a possible neuroprotectant was evaluated.

Methods : Oxidative stress was induced by adding 300 µM H2O2 on day one for three hours to porcine retinal organotypic cultures. At the same time, 500 µM iNOS-inhibitor (1400W, Merck Millipore) was added. Neuronal and glial cells were evaluated via qRT-PCR and immunohistology after four and eight days. In addition, TEM was performed. The following groups were compared: controls, H2O2, and H2O2+iNOS-inhibitor.

Results : H2O2-induced retinal ganglion cell (RGC) loss (4 days: p=0.0001; 8 days: p=0.005) was prevented by iNOS-inhibitor treatment (4 days: p=0005; 8 days: p=0.13). On mRNA level, a significantly increased expression of autophagic gene p62 due to H2O2 (p=0.037) was prevented by the iNOS-inhibitor treatment after four days (p=0.45). Likewise, a reduced expression of caspase 8 was seen after iNOS-inhibitor treatment (3.19-fold; p=0.54) in comparison to the H2O2-stressed group (5.37-fold; p=0.065). A late rescue of bipolar cells was noted in iNOS-inhibitor treated retinas (p=0.0001). TEM revealed that cell morphology as well as the cell compartments of RGCs were protected from oxidative stress damage by the iNOS-inhibitor.

Conclusions : Strong degeneration in porcine retinas due to H2O2-induced oxidative stress was prevented by treatment with the iNOS-inhibitor. A lowered apoptotic stage resulted in a cell rescue, especially of RGCs. These results indicate, a potential neuroprotective role of the iNOS-inhibitor 1400W in diseases affecting the inner retina, especially glaucoma or retinal ischemia.

This is a 2020 ARVO Annual Meeting abstract.

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