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101 EFFECTS OF ANTIOXIDANTS ON TERT-BUTYL HYDROPEROXIDE CELL KILLING IN MACROPHAGE (RAW 264.7) CELLS.
  1. J. B. Bullock1,
  2. P. O. Obih1
  1. 1College of Pharmacy, Xavier University of Louisiana, New Orleans, LA.

Abstract

The aim of this study was to examine the protective effects of a variety of antioxidants, including alpha-tocopherol (vitamin E), ascorbic acid, and selenium, in an in vitro model of alcohol-mediated free radical generation in the liver, one of the major organs affected by chronic alcohol abuse. Hepatic macrophage cells (RAW 264.7 cells) were cultured to confluence in 24-well plates. A dose-response curve of tBH (tert-butyl hydroperoxide) killing in the presence or absence of ethanol (EtOH) was determined. Cell killing was evaluated by determining cell leakage of lactate dehydrogenase. Cells were then exposed to a tBH dose that killed 50% of the cells in the presence of various doses of each of the potential antioxidants: ascorbic acid, alpha-tocopherol, or selenium. Alpha-tocopherol acted as an antioxidant by decreasing tBH cell killing in a dose-dependent manner. Selenium at a relatively high dose (10 μmol had no effect on tBH killing. However, at 50 μmol, it enhanced tBH cell killing, thereby acting as a pro-oxidant. Ascorbic acid at low doses protected the cells from tBH cell killing, but at doses higher than 500 μmol, it enhanced cell killing, thereby acting as a pro-oxidant. Our results indicate that protection from free radical damage depends on the dose of alpha-tocopherol, ascorbic acid, or selenium. An antioxidant regimen can be developed to minimize free radical generation due to ethanol metabolism that will significantly reduce the subsequent damage to the liver caused by alcohol abuse.

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