Free fatty acids enhance the oxidative damage induced by ethanol metabolism in an in vitro model

Food Chem Toxicol. 2015 Feb:76:109-15. doi: 10.1016/j.fct.2014.12.005. Epub 2014 Dec 17.

Abstract

In recent years, there has been a growing interest to explore the responsiveness to injury in steatotic hepatocyte. VL-17A cells, which express ADH and Cyp2E1 overloaded with free fatty acids (1 mM of oleic and palmitic acid 2:1) showed an increased oxidative damaged after 24 h free fatty acids treatment when exposed to ethanol (100 mM) for 48 h as a second injury. An increment in reactive oxygen species, determined by DCFH-DA, protein oxidation, and apoptosis were observed although an increase in main antioxidant proteins such as superoxide dismutase 1 and glutathione peroxidase were observed, but failed in gamma-glutamylcysteine synthetase, suggesting a decreased capacity of synthesis of glutathione compared with cells treated only with free fatty acids or ethanol. The increased oxidative stress and toxicity in lipid overloaded VL-17A cells subjected to ethanol exposure were accompanied by increases in Cyp2E1 protein expression. Our data show that lipid loaded in an in vitro model, VL-17A cells, is more susceptible to cell damage and oxidative stress when treated with ethanol.

Keywords: Ethanol; Hepatocytes; Oxidative stress; Steatosis; VL-17A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Dehydrogenase / metabolism
  • Antioxidants / pharmacology
  • Apoptosis / drug effects
  • Cytochrome P-450 CYP2E1 / genetics
  • Cytochrome P-450 CYP2E1 / metabolism
  • Ethanol / toxicity*
  • Fatty Acids, Nonesterified / pharmacology*
  • Fluoresceins / metabolism
  • Glutamate-Cysteine Ligase / metabolism
  • Glutathione / biosynthesis
  • Hep G2 Cells
  • Hepatocytes / drug effects
  • Hepatocytes / metabolism
  • Humans
  • Oxidative Stress / drug effects*
  • Reactive Oxygen Species / metabolism
  • Triglycerides / metabolism

Substances

  • Antioxidants
  • Fatty Acids, Nonesterified
  • Fluoresceins
  • Reactive Oxygen Species
  • Triglycerides
  • diacetyldichlorofluorescein
  • Ethanol
  • Alcohol Dehydrogenase
  • Cytochrome P-450 CYP2E1
  • Glutamate-Cysteine Ligase
  • Glutathione