Effects of the Selective EP2 Receptor Agonist Omidenepag on Adipocyte Differentiation in 3T3-L1 Cells

J Ocul Pharmacol Ther. 2020 Apr;36(3):162-169. doi: 10.1089/jop.2019.0079. Epub 2020 Jan 14.

Abstract

Purpose: We aimed at comparing the effects of omidenepag (OMD) with those of prostaglandin F (FP) receptor agonists (FP agonists) on adipogenesis in mouse 3T3-L1 cells. Methods: To evaluate the agonistic activities of OMD against the mouse EP2 (mEP2) receptor, we determined cAMP contents in mEP2 receptor-expressing CHO cells by using radioimmunoassays. Overall, 3T3-L1 cells were cultured in differentiation medium for 10 days and adipocyte differentiation was assessed according to Oil Red O-stained cell areas. Changes in expression levels of the adipogenic transcription factors Pparg, Cebpa, and Cebpb were determined by using real-time polymerase chain reaction (PCR). OMD at 0.1, 1, 10, and 40 μmol/L, latanoprost free acid (LAT-A) at 0.1 μmol/L, or prostaglandin F (PGF), at 0.1 μmol/L were added to cell culture media during adipogenesis. Oil Red O-stained areas and expression patterns of transcription factor targets of OMD or FP agonists were compared with those of untreated controls. Results: The 50% effective concentration (EC50) of OMD against the mEP2 receptor was 3.9 nmol/L. Accumulations of Oil Red O-stained lipid droplets were observed inside control cells on day 10. LAT-A and PGF significantly inhibited the accumulation of lipid droplets; however, OMD had no effect on this process even at concentrations up to 40 μmol/L. LAT-A and PGF significantly suppressed Pparg, Cebpa, and Cebpb gene expression levels during adipocyte differentiation. Conversely, OMD had no obvious effects on the expression levels of these genes. Conclusions: A selective EP2 receptor agonist, OMD, did not affect the adipocyte differentiation in 3T3-L1 cells, whereas FP agonists significantly inhibited this process.

Keywords: EP2 agonist; FP agonist; adipogenesis; deepening of the upper eyelid sulcus; omidenepag.

Publication types

  • Comparative Study

MeSH terms

  • 3T3-L1 Cells / drug effects*
  • 3T3-L1 Cells / metabolism
  • Adipocytes / cytology
  • Adipocytes / drug effects*
  • Adipocytes / metabolism
  • Adipogenesis / drug effects*
  • Animals
  • CHO Cells / drug effects
  • CHO Cells / metabolism
  • Cell Differentiation / drug effects
  • Cricetulus
  • Cyclic AMP / metabolism
  • Disease Models, Animal
  • Glycine / analogs & derivatives*
  • Glycine / pharmacology
  • Latanoprost / pharmacology*
  • Lipid Droplets / drug effects
  • Lipid Droplets / metabolism
  • Mice
  • Prostaglandins F, Synthetic / pharmacology
  • Pyrazoles / pharmacology*
  • Pyridines / pharmacology*
  • Radioimmunoassay / methods
  • Receptors, Prostaglandin E, EP2 Subtype / agonists*

Substances

  • Prostaglandins F, Synthetic
  • Pyrazoles
  • Pyridines
  • Receptors, Prostaglandin E, EP2 Subtype
  • Latanoprost
  • Cyclic AMP
  • omidenepag isopropyl
  • Glycine