Ethanol metabolism and oxidative stress are required for unfolded protein response activation and steatosis in zebrafish with alcoholic liver disease

Tsedensodnom, O., Vacaru, A.M., Howarth, D.L., Yin, C., and Sadler, K.C.
Disease models & mechanisms   6(5): 1213-26 (Journal)
Registered Authors
Howarth, Deanna, Sadler Edepli, Kirsten C., Tsedensodnom, OonoO, Vacaru, Ana, Yin, Chunyue
MeSH Terms
  • Alcohol Dehydrogenase/metabolism
  • Animals
  • Antioxidants/pharmacology
  • Cytochrome P-450 CYP2E1/metabolism
  • Endoplasmic Reticulum Stress/drug effects
  • Ethanol/metabolism*
  • Ethanol/toxicity
  • Fatty Liver/complications*
  • Fatty Liver/metabolism*
  • Hepatic Stellate Cells/drug effects
  • Hepatic Stellate Cells/metabolism
  • Hepatic Stellate Cells/pathology
  • Larva/drug effects
  • Larva/metabolism
  • Liver/drug effects
  • Liver/metabolism
  • Liver/pathology
  • Liver Diseases, Alcoholic/complications*
  • Liver Diseases, Alcoholic/metabolism
  • Oxidative Stress*/drug effects
  • Reactive Oxygen Species/metabolism
  • Secretory Pathway/drug effects
  • Survival Analysis
  • Unfolded Protein Response*/drug effects
  • Zebrafish/metabolism*
23798569 Full text @ Dis. Model. Mech.

Secretory pathway dysfunction and lipid accumulation (steatosis) are the two most common responses of hepatocytes exposed to ethanol and are major factors in the pathophysiology of alcoholic liver disease (ALD). However, the mechanisms by which ethanol elicits these cellular responses are not fully understood. Recent data indicates that activation of the unfolded protein response (UPR) in response to secretory pathway dysfunction can cause steatosis. Here, we examined the relationship between alcohol metabolism, oxidative stress, secretory pathway stress and steatosis using zebrafish larvae. We found that ethanol was immediately internalized and metabolized by larvae, such that the internal ethanol concentration in 4 day old larvae equilibrated to 160 mM after 1 hour of exposure to 350 mM ethanol, with an average ethanol metabolism rate of 56 µmol/larva/hour over 32 hours. Blocking alcohol dehydrogenase 1 (Adh1) and cytochrome P450 2E1 (Cyp2e1), the major enzymes that metabolize ethanol, prevented alcohol-induced steatosis and reduced induction of the UPR in the liver. Thus, we conclude that ethanol metabolism causes ALD in zebrafish. Oxidative stress generated by Cyp2e1-mediated ethanol metabolism is proposed to be a major culprit in ALD pathology. We found that production of reactive oxygen species (ROS) increased in larvae exposed to ethanol, while either inhibition of the zebrafish Cyp2e1 homolog or administration of antioxidants reduced ROS levels. Importantly, these treatments also blocked ethanol-induced steatosis and reduced UPR activation, while H2O2 acted as a pro-oxidant that synergized with low doses of ethanol to induce the UPR. Collectively, these data demonstrate that ethanol metabolism and oxidative stress are conserved mechanisms required for the development of steatosis and hepatic dysfunction in ALD, and that these processes contribute to ethanol-induced UPR activation and secretory pathway stress in hepatocytes.

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