The role of AMP-activated protein kinase in the action of ethanol in the liver.
OBJECTIVE
Our previous work has shown that ethanol induces the fatty acid synthesis pathway by activation of sterol regulatory element-binding protein 1 (SREBP-1). In the present study, we studied the mechanisms of this activation by identifying a new target of ethanol, AMP-activated protein kinase (AMPK).


METHODS
The effects of ethanol on AMPK, acetyl-CoA carboxylase (ACC), and SREBP-1 were assessed in rat hepatic cells and in the livers of ethanol-fed mice.


RESULTS
In rat hepatoma H4IIEC3 or McA-RH 7777 cell lines, ethanol-induced transcription of an SREBP-regulated promoter was suppressed by the presence of 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) or metformin, 2 known AMPK activators. Consistent with this, over expression of a constitutively active form of AMPK blocked the effect of ethanol, whereas coexpression of a dominant-negative form of AMPK augmented the effect. Moreover, activation of AMPK by metformin or AICAR largely blocked the ability of ethanol to increase levels of mature SREBP-1 protein. These findings suggest that the effect of ethanol on SREBP-regulated promoter activation was partially mediated through AMPK inhibition. We further demonstrated that AMPK was inhibited by ethanol in hepatic cells. In parallel, ethanol increased the activity of ACC and suppressed the rate of palmitic acid oxidation. Finally, feeding mice a low-fat diet with ethanol resulted in significantly reduced hepatic AMPK activity, increased ACC activity, and enhanced malonyl CoA content.


CONCLUSIONS
Taken together, our findings suggest that AMPK may play a key role in regulating the effects of ethanol on SREBP-1 activation, fatty acid metabolism, and development of alcoholic fatty liver.
You
Min
M
Depatment of Medicine, Indiana University School of Medicine, Richard Roudebush Veteran's Affairs Medical Center, Indianapolis, Indiana 46202, USA.
Matsumoto
Michinaga
M
Pacold
Christine M
CM
Cho
Won Kyoo
WK
Crabb
David W
DW
D055372:AMP-Activated Protein Kinases; D000818:Animals; D022762:CCAAT-Enhancer-Binding Proteins / Q000494:pharmacology*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D002492:Central Nervous System Depressants / Q000009:adverse effects*; D004268:DNA-Binding Proteins / Q000494:pharmacology*; D000431:Ethanol / Q000009:adverse effects*; D005227:Fatty Acids / Q000378:metabolism; D005235:Fatty Liver, Alcoholic / Q000503:physiopathology; D005786:Gene Expression Regulation; D018257:Helix-Loop-Helix Motifs; D022781:Hepatocytes / Q000201:enzymology*; D006801:Humans; D016350:Leucine Zippers; D008114:Liver Neoplasms, Experimental / Q000201:enzymology / Q000503:physiopathology; D051379:Mice; D009097:Multienzyme Complexes / Q000502:physiology*; D010084:Oxidation-Reduction; D011401:Promoter Regions, Genetic; D017346:Protein Serine-Threonine Kinases / Q000502:physiology*; D051381:Rats; D051780:Sterol Regulatory Element Binding Protein 1; D014157:Transcription Factors / Q000494:pharmacology*
D022762:CCAAT-Enhancer-Binding Proteins; D002492:Central Nervous System Depressants; D004268:DNA-Binding Proteins; D005227:Fatty Acids; D009097:Multienzyme Complexes; C497387:SREBF1 protein, human; C497388:Srebf1 protein, mouse; C497389:Srebf1 protein, rat; D051780:Sterol Regulatory Element Binding Protein 1; D014157:Transcription Factors; D000431:Ethanol; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases