April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
CYP2E1 mediated ethanol metabolism in ARPE-19 cells.
Author Affiliations & Notes
  • Natalia Martínez
    Phisiology, Universidad Católica de Valencia, Valencia, Spain
  • Sandra Atienzar
    Phisiology, Universidad Católica de Valencia, Valencia, Spain
  • Miguel Flores-Bellver
    Phisiology, Universidad Católica de Valencia, Valencia, Spain
  • Luis Bonet
    Phisiology, Universidad Católica de Valencia, Valencia, Spain
  • Javier Sancho-Pelluz
    Phisiology, Universidad Católica de Valencia, Valencia, Spain
  • Jorge M Barcia
    Phisiology, Universidad Católica de Valencia, Valencia, Spain
  • Francisco J Romero
    Phisiology, Universidad Católica de Valencia, Valencia, Spain
  • Footnotes
    Commercial Relationships Natalia Martínez, None; Sandra Atienzar, None; Miguel Flores-Bellver, None; Luis Bonet, None; Javier Sancho-Pelluz, None; Jorge Barcia, None; Francisco Romero, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 6322. doi:https://doi.org/
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      Natalia Martínez, Sandra Atienzar, Miguel Flores-Bellver, Luis Bonet, Javier Sancho-Pelluz, Jorge M Barcia, Francisco J Romero; CYP2E1 mediated ethanol metabolism in ARPE-19 cells.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):6322. doi: https://doi.org/.

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

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Abstract

Purpose: Alcohol consumption has a high prevalence in most countries and it is the drug with the highest use and abuse worldwide. Chronic ethanol consumption produces an increase in lipid peroxidation products (MDA) and a decrease in antioxidant factors, (GSH) and its related enzymes, which can eventually induce apoptosis-mediated cell death on central nervous system. Retina is particularly sensitive to free radicals (ROS) and lipid peroxidation. Pigmented epithelial (RPE) cells play a major role in retinal homeostasis. There are few studies on ethanol-induced RPE injury related to oxidative stress. We used ARPE-19 cells to study if there is ethanol metabolism via CYP2E1 (P450) in retinal pigment epithelium.

Methods: The presence of P450 in ARPE-19 cells was studied by immunocytochemistry and Western Blot. Cell viability and apoptosis were studied by MTT and Western Blot. Regulation of CYP2E1 expression by ethanol-induced oxidative stress was studied by Western Blot, immunocytochemistry and RT-PCR, using a CYP2E1 selective inhibitor, Diallyl Sulfide (DAS). CYP2E1 activity was measured by colorimetric assay. The involvement of CYP2E1 in oxidative stress-mediated apoptosis and/or autophagy was analyzed by Western Blot, ELISA, and a proteome profiler cell stress assay.

Results: We show for the first time the presence of CYP2E1 in RPE. Its expression was induced by ethanol and was concentration dependent. DAS treatment decreased the expression of the CYP2E1 and the amount of mRNA, reducing the formation of ROS in cultured cells.

Conclusions: CYP2E1 is present in retinal pigment epithelium and metabolizes ethanol; it is inducible on a concentration-dependent manner and DAS is able to inhibit its expression, reducing the formation of ROS.

Keywords: 592 metabolism • 726 stress response • 701 retinal pigment epithelium  
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