Oleuropein and hydroxytyrosol inhibit adipocyte differentiation in 3 T3-L1 cells

Life Sci. 2011 Nov 7;89(19-20):708-16. doi: 10.1016/j.lfs.2011.08.012. Epub 2011 Sep 10.


Aims: Oleuropein and hydroxytyrosol, which are antioxidant molecules found in olive leaves and oil, have been reported to exert several biochemical and pharmacological effects. These polyphenols are able to prevent low-density lipoprotein oxidation and protect cells against several diseases. Here, we studied the effect of these compounds on adipocyte differentiation in 3 T3-L1.

Main methods: To perform this study, 3 T3-L1 preadipocytes viability was analysed via Trypan blue and MTT assays, and triglycerides were stained with Oil Red O. Adipogenesis related genes expression were checked by RT-PCR and qRT-PCR. Also, cells counting and flow cytometry were used to analyse the mitotic cell cycle during the adipogenesis clonal expansion phase.

Results: Oleuropein and hydroxytyrosol dose-dependently suppressed intracellular triglyceride accumulation during adipocyte differentiation without effect on cell viability. PPARγ, C/EBPα and SREBP-1c transcription factors and their downstream targets genes (GLUT4, CD36 and FASN) were down-regulated after treatment by oleuropein and hydroxytyrosol. At 200 and 300 μmol/L oleuropein or 100 and 150 μmol/L hydroxytyrosol, the greatest effect on the adipogenesis process was observed during the early stages of differentiation. Flow cytometry revealed both polyphenols to inhibit the division of 3T3-L1 preadipocytes during mitotic clonal expansion and cause cell cycle delay. Furthermore, oleuropein and its derivate hydroxytyrosol decreased the transcriptional activity of SREBP-1c in a stable transfected 3T3-L1 cell line.

Significance: These findings indicate that both compounds are able to prevent 3T3-L1 differentiation by inhibition of the mitotic clonal expansion and downregulation of the adipogenesis related genes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3-L1 Cells
  • Adipocytes / drug effects*
  • Adipocytes / metabolism
  • Adipogenesis / drug effects
  • Adipogenesis / genetics
  • Animals
  • Antioxidants / administration & dosage
  • Antioxidants / pharmacology*
  • Cell Differentiation / drug effects*
  • Cell Survival / drug effects
  • Dose-Response Relationship, Drug
  • Down-Regulation / drug effects
  • Flow Cytometry
  • Iridoid Glucosides
  • Iridoids
  • Mice
  • Mitosis / drug effects
  • Phenylethyl Alcohol / administration & dosage
  • Phenylethyl Alcohol / analogs & derivatives*
  • Phenylethyl Alcohol / pharmacology
  • Pyrans / administration & dosage
  • Pyrans / pharmacology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Triglycerides / metabolism


  • Antioxidants
  • Iridoid Glucosides
  • Iridoids
  • Pyrans
  • Triglycerides
  • 3,4-dihydroxyphenylethanol
  • oleuropein
  • Phenylethyl Alcohol