Diazinon exposure activated transcriptional factors CCAAT-enhancer-binding proteins α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ) and induced adipogenesis in 3T3-L1 preadipocytes

Pestic Biochem Physiol. 2018 Sep:150:48-58. doi: 10.1016/j.pestbp.2018.07.003. Epub 2018 Jul 10.

Abstract

Environmental chemical exposure could be a contributor to the increasing obesity epidemic. Diazinon, an organophosphate insecticide, has been widely used in the agriculture, and exposure of the general population to diazinon has been reported. Diazinon has been known to induce neurotoxic effects mainly through the inhibition of acetylcholinesterase (AChE). However, its association with dysregulation of adipogenesis has been poorly investigated. The current study aimed to examine the mechanism of diazinon's effect on adipogenesis using the 3T3-L1 preadipocytes combined with a single-cell-based high-content analysis. The results showed that diazinon induced lipid droplet accumulation in a dose-dependent manner. The dynamic changes of adipogenic regulatory proteins and genes were examined at the three stages of adipogenesis (induction, differentiation, and maturation) in 3T3-L1 cells treated with various doses of diazinon (0, 1, 10, 100 μM) using real-time quantitative RT-PCR and Western Blot respectively. Diazinon significantly induced protein expression of transcriptional factors CCAAT-enhancer-binding proteins α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ), their downstream proteins, fatty acid synthase (FASN), acetyl CoA carboxylase (ACC), fatty acid-binding protein 4 (FABP4), lipoprotein lipase (LPL), adiponectin and perilipin in dose and time-dependent manners. Similarly, the adipogenic genes were significantly induced in a dose and time-dependent manner compared to the relative controls. The current study demonstrates that diazinon promotes lipid accumulation and activates the adipogenic signaling pathway in the in vitro model.

Keywords: 3T3-L1; Adipogenesis; Diazinon; Obesogen.

MeSH terms

  • 3T3-L1 Cells
  • Acetyl-CoA Carboxylase / metabolism
  • Adipogenesis / drug effects*
  • Adiponectin / metabolism
  • Animals
  • Blotting, Western
  • CCAAT-Enhancer-Binding Protein-alpha / metabolism*
  • Cholinesterase Inhibitors / pharmacology*
  • Diazinon / pharmacology*
  • Dose-Response Relationship, Drug
  • Fatty Acid Synthases / metabolism
  • Fatty Acid-Binding Proteins / metabolism
  • Insecticides / pharmacology*
  • Lipid Metabolism
  • Lipoprotein Lipase / metabolism
  • Mice
  • PPAR gamma / metabolism*
  • Perilipin-1 / metabolism
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction / drug effects

Substances

  • Adiponectin
  • CCAAT-Enhancer-Binding Protein-alpha
  • Cholinesterase Inhibitors
  • Fabp4 protein, mouse
  • Fatty Acid-Binding Proteins
  • Insecticides
  • PPAR gamma
  • Perilipin-1
  • Fatty Acid Synthases
  • Lipoprotein Lipase
  • Acetyl-CoA Carboxylase
  • Diazinon