title: Docosahexaenoic acid suppresses apolipoprotein A-I gene expression through hepatocyte nuclear factor-3β.
abstract: BACKGROUND
  Dietary fish-oil supplementation has been shown in human kinetic studies to lower the production rate of apolipoprotein (apo) A-I, the major protein component of HDL. The underlying mechanism responsible for this effect is not fully understood.
  
  
  OBJECTIVE
  We investigated the effect and the mechanism of action of the very-long-chain n-3 (omega-3) polyunsaturated fatty acid docosahexaenoic acid (DHA), relative to the saturated fatty acid palmitic acid (PA), on the hepatic expression of apo A-I in HepG2 cells.
  
  
  METHODS
  HepG2 cells were treated with different doses of DHA and PA (0-200 μmol/L). mRNA expression levels of apo A-I were assessed by real-time polymerase chain reaction, and apo A-I protein concentrations were measured by immunoassay. DHA dose-dependently suppressed apo A-I mRNA levels and also lowered apo A-I protein concentrations in the media, with maximum effects at 200 μmol/L. This concentration of fatty acids was used in all subsequent experiments.
  
  
  RESULTS
  To elucidate the mechanism mediating the reduction in apo A-I expression by DHA, transfection experiments were conducted with plasmid constructs containing serial deletions of the apo A-I promoter. The DHA-responsive region was mapped to the -185 to -148 nucleotide region of the apo A-I promoter, which binds the hepatocyte nuclear factor (HNF)-3β. Nuclear extracts from cells treated with DHA or PA had a similar nuclear abundance of HNF-3β. However, electrophoresis mobility shift assays showed less binding of HNF-3β to the -180 to -140 sequence of the apo A-I promoter than did PA-treated cells. As shown by chromatin immunoprecipitation analysis, less HNF-3β was recruited to the apo A-I promoter in DHA-treated cells than in PA-treated cells, which supports the concept of an interference of DHA with the binding of HNF-3β to the apo A-I promoter.
  
  
  CONCLUSIONS
  These findings suggest that, in human hepatoma HepG2 cells, DHA inhibits the binding of HNF-3β to the apo A-I promoter, resulting in the repression of apo A-I promoter transactivity and thus a reduction in apo A-I expression.
authors.affiliation: Lipid Metabolism Laboratory, Jean Mayer US Department of Agriculture Human Nutrition Research Center on Aging at Tufts University, Boston, MA, USA.
authors.forename: Yu-Lin
authors.initials: YL
authors.lastname: Kuang
authors.forename: K Eric
authors.initials: KE
authors.lastname: Paulson
authors.forename: Alice H
authors.initials: AH
authors.lastname: Lichtenstein
authors.forename: Nirupa R
authors.initials: NR
authors.lastname: Matthan
authors.forename: Stefania
authors.initials: S
authors.lastname: Lamon-Fava
mesh_terms: D016632:Apolipoprotein A-I / Q000235:genetics* / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D004281:Docosahexaenoic Acids / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D056945:Hep G2 Cells; D051542:Hepatocyte Nuclear Factor 3-beta / Q000502:physiology*; D006801:Humans; D008099:Liver / Q000378:metabolism; D011401:Promoter Regions, Genetic
chemical_list: D016632:Apolipoprotein A-I; D051542:Hepatocyte Nuclear Factor 3-beta; D004281:Docosahexaenoic Acids