Benzyl butyl phthalate promotes adipogenesis in 3T3-L1 preadipocytes: A High Content Cellomics and metabolomic analysis

Toxicol In Vitro. 2016 Apr;32:297-309. doi: 10.1016/j.tiv.2016.01.010. Epub 2016 Jan 25.

Abstract

Benzyl butyl phthalate (BBP) has been known to induce developmental and reproductive toxicity. However, its association with dysregulation of adipogenesis has been poorly investigated. The present study aimed to examine the effect of BBP on the adipogenesis, and to elucidate the underlying mechanisms using the 3T3-L1 cells. The capacity of BBP to promote adipogenesis was evaluated by multiple staining approaches combined with a High Content Cellomics analysis. The dynamic changes of adipogenic regulatory genes and proteins were examined, and the metabolite profile was identified using GC/MC based metabolomic analysis. The High Content analysis showed BBP in contrast with Bisphenol A (BPA), a known environmental obesogen, increased lipid droplet accumulation in a similar dose-dependent manner. However, the size of the lipid droplets in BBP-treated cells was significantly larger than those in cells treated with BPA. BBP significantly induced mRNA expression of transcriptional factors C/EBPα and PPARγ, their downstream genes, and numerous adipogenic proteins in a dose and time-dependent manner. Furthermore, GC/MC metabolomic analysis revealed that BBP exposure perturbed the metabolic profiles that are associated with glyceroneogenesis and fatty acid synthesis. Altogether, our current study clearly demonstrates that BBP promoted the differentiation of 3T3-L1 through the activation of the adipogenic pathway and metabolic disturbance.

Keywords: Adipogenesis; Benzyl butyl phthalate; Gene expression; High Content Cellomics analysis (HCA); Metabolomic analysis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3-L1 Cells
  • Adipocytes / drug effects
  • Adipocytes / metabolism
  • Adipogenesis / drug effects*
  • Adiponectin / metabolism
  • Animals
  • CCAAT-Enhancer-Binding Protein-alpha / genetics
  • CCAAT-Enhancer-Binding Protein-alpha / metabolism
  • Cell Differentiation / drug effects
  • Complement Factor D / metabolism
  • Fatty Acid Synthase, Type I / metabolism
  • Fatty Acid-Binding Proteins / metabolism
  • Lipid Metabolism / drug effects
  • Lipoprotein Lipase / metabolism
  • Metabolomics
  • Mice
  • PPAR gamma / genetics
  • PPAR gamma / metabolism
  • Phthalic Acids / pharmacology*
  • RNA, Messenger / metabolism

Substances

  • Adiponectin
  • Adipoq protein, mouse
  • CCAAT-Enhancer-Binding Protein-alpha
  • Fabp4 protein, mouse
  • Fatty Acid-Binding Proteins
  • PPAR gamma
  • Phthalic Acids
  • RNA, Messenger
  • Fatty Acid Synthase, Type I
  • Lipoprotein Lipase
  • Complement Factor D
  • complement factor D, mouse
  • butylbenzyl phthalate