title: Ethanol promotes thiamine deficiency-induced neuronal death: involvement of double-stranded RNA-activated protein kinase.
abstract: BACKGROUND
  Heavy alcohol consumption causes cerebellar degeneration, and the underlying mechanism is unclear. Chronic alcoholism is usually associated with thiamine deficiency (TD) which is known to induce selective neurodegeneration in the brain. However, the role of TD in alcohol-induced cerebellar degeneration remains to be elucidated. The double-stranded RNA-activated protein kinase (PKR) is a potent antiviral protein. Viral infection or binding to dsRNA causes PKR autophosphorylation and subsequent phosphorylation of the alpha-subunit of eukaryotic translation factor-2alpha, leading to inhibition of translation or apoptosis. PKR can also be activated by cellular stresses.
  
  
  METHODS
  In this study, we used an in vitro model, cultured cerebellar granule neurons (CGNs), to investigate the interaction between TD and ethanol and evaluate the contribution of their interaction to neuronal loss. TD was induced by treatment with amprolium in association with ethanol. Cell viability was determined by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide assay. PKR expression/phosphorylation and subcellular distribution was analyzed with immunoblotting and immunocytochemistry.
  
  
  RESULTS
  Thiamine deficiency caused death of CGNs but ethanol did not. However, TD plus ethanol induced a much greater cell loss than TD alone. TD-induced PKR phosphorylation and ethanol exposure significantly promoted TD-induced PKR phosphorylation as well as its nuclear translocation. A selective PKR inhibitor not only protected CGNs against TD toxicity, but also abolished ethanol potentiation of TD-induced loss of CGNs.
  
  
  CONCLUSIONS
  Ethanol promoted TD-induced PKR activation and neuronal death. PKR may be a convergent protein that mediates the interaction between TD and ethanol.
authors.affiliation: Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Zun-Ji
authors.initials: ZJ
authors.lastname: Ke
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Zhiqin
authors.initials: Z
authors.lastname: Fan
authors.forename: Jia
authors.initials: J
authors.lastname: Luo
mesh_terms: D000437:Alcoholism / Q000378:metabolism; D000670:Amprolium / Q000494:pharmacology; D000818:Animals; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000633:toxicity*; D003049:Coccidiostats / Q000494:pharmacology; D004195:Disease Models, Animal; D000431:Ethanol / Q000633:toxicity*; D009410:Nerve Degeneration / Q000209:etiology* / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013831:Thiamine / Q000378:metabolism; D013832:Thiamine Deficiency / Q000150:complications* / Q000378:metabolism; D019892:eIF-2 Kinase / Q000378:metabolism*
chemical_list: D002492:Central Nervous System Depressants; D003049:Coccidiostats; D000431:Ethanol; D019892:eIF-2 Kinase; D000670:Amprolium; D013831:Thiamine