HIF-1α expression as a protective strategy of HepG2 cells against fatty acid-induced toxicity

J Cell Biochem. 2014 Jun;115(6):1147-58. doi: 10.1002/jcb.24757.

Abstract

Free fatty acid-induced lipotoxicity via increased endoplasmic reticulum (ER) stress and hepatocyte apoptosis is a key pathological mechanism of non-alcoholic steatohepatitis. A role of hypoxia-inducible factor 1α (HIF-1α) in this process has been suggested, but direct evidence is lacking. Here, we used HepG2 cells as a model to study whether HIF-1α can reduce palmitic acid-induced lipotoxicity and ER stress. In HepG2 cells treated with 500 µM palmitic acid, HIF-1α expression increased transiently, the decline was associated with increased cleaved caspase-3 expression. Overexpression and knockdown of HIF-1α decreased and exacerbated, respectively, palmitic acid-induced lipoapoptosis. The overexpression also blunted upregulation of the ER stress markers, C/EBP homologous protein (CHOP) and chaperone immunoglobulin heavy chain binding protein (Bip), while the knockdown increased the level of CHOP. In line with this, CHOP promoter activity decreased following HIF-1α binding to the CHOP promoter hypoxia response element. These results indicate that hepatocyte lipotoxicity is associated with decreased HIF-1α expression. It also suggests that upregulation of HIF-1α can be a possible strategy to reduce lipotoxicity in non-alcoholic fatty liver disease.

Keywords: CHOP; ER STRESS; HEPATIC STEATOSIS; HIF-1α; LIPOAPOPTOSIS; NON-ALCOHOLIC FATTY LIVER DISEASE.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects*
  • Blotting, Western
  • Caspase 3 / metabolism
  • Cell Hypoxia
  • Cell Survival / drug effects
  • Choline Deficiency
  • Diet
  • Endoplasmic Reticulum Chaperone BiP
  • Endoplasmic Reticulum Stress / drug effects
  • Endoplasmic Reticulum Stress / genetics
  • Gene Expression / drug effects
  • Heat-Shock Proteins / genetics
  • Heat-Shock Proteins / metabolism
  • Hep G2 Cells
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / genetics
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism*
  • Liver / metabolism
  • Liver / pathology
  • Methionine / deficiency
  • Mice, Inbred C57BL
  • Palmitic Acid / pharmacology*
  • Promoter Regions, Genetic / genetics
  • Protective Factors
  • Protein Binding
  • RNA Interference
  • Response Elements / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factor CHOP / genetics
  • Transcription Factor CHOP / metabolism

Substances

  • DDIT3 protein, human
  • Endoplasmic Reticulum Chaperone BiP
  • HIF1A protein, human
  • Heat-Shock Proteins
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Transcription Factor CHOP
  • Palmitic Acid
  • Methionine
  • Caspase 3