Synergistic suppression of early phase of adipogenesis by microsomal PGE synthase-1 (PTGES1)-produced PGE2 and aldo-keto reductase 1B3-produced PGF2α

PLoS One. 2012;7(9):e44698. doi: 10.1371/journal.pone.0044698. Epub 2012 Sep 7.

Abstract

We recently reported that aldo-keto reductase 1B3-produced prostaglandin (PG) F(2α) suppressed the early phase of adipogenesis. PGE(2) is also known to suppress adipogenesis. In this study, we found that microsomal PGE(2) synthase (PGES)-1 (mPGES-1; PTGES1) acted as the PGES in adipocytes and that PGE(2) and PGF(2α) synergistically suppressed the early phase of adipogenesis. PGE(2) production was detected in preadipocytes and transiently enhanced at 3 h after the initiation of adipogenesis of mouse adipocytic 3T3-L1 cells, followed by a quick decrease; and its production profile was similar to the expression of the cyclooxygenase-2 (PTGS2) gene. When 3T3-L1 cells were transfected with siRNAs for any one of the three major PTGESs, i.e., PTGES1, PTGES2 (mPGES-2), and PTGES3 (cytosolic PGES), only PTGES1 siRNA suppressed PGE(2) production and enhanced the expression of adipogenic genes. AE1-329, a PTGER4 (EP4) receptor agonist, increased the expression of the Ptgs2 gene with a peak at 1 h after the initiation of adipogenesis. PGE(2)-mediated enhancement of the PTGS2 expression was suppressed by the co-treatment with L-161982, a PTGER4 receptor antagonist. Moreover, AE1-329 enhanced the expression of the Ptgs2 gene by binding of the cyclic AMP response element (CRE)-binding protein to the CRE of the Ptgs2 promoter; and its binding was suppressed by co-treatment with L-161982, which was demonstrated by promoter luciferase and chromatin immunoprecipitation assays. Furthermore, when 3T3-L1 cells were caused to differentiate into adipocytes in medium containing both PGE(2) and PGF(2α), the expression of the adipogenic genes and the intracellular triglyceride level were decreased to a greater extent than in medium containing either of them, revealing that PGE(2) and PGF(2α) independently suppressed adipogenesis. These results indicate that PGE(2) was synthesized by PTGES1 in adipocytes and synergistically suppressed the early phase of adipogenesis of 3T3-L1 cells in cooperation with PGF(2α) through receptor-mediated activation of PTGS2 expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3-L1 Cells
  • Adipocytes / cytology*
  • Animals
  • Base Sequence
  • Blotting, Western
  • Chromatin Immunoprecipitation
  • DNA Primers
  • Dinoprost / biosynthesis*
  • Dinoprostone / biosynthesis*
  • Hydroxyprostaglandin Dehydrogenases / metabolism*
  • Intramolecular Oxidoreductases / metabolism*
  • Mice
  • Microsomes / enzymology*
  • Prostaglandin-E Synthases
  • RNA Interference
  • RNA, Small Interfering

Substances

  • DNA Primers
  • RNA, Small Interfering
  • Dinoprost
  • Hydroxyprostaglandin Dehydrogenases
  • prostaglandin-F synthase
  • Intramolecular Oxidoreductases
  • Prostaglandin-E Synthases
  • Ptges protein, mouse
  • Ptges2 protein, mouse
  • Ptges3 protein, mouse
  • Dinoprostone

Grant support

This work was supported in part by Grant-in-Aid for Scientific Research (21570151)and Scientific Research on Innovative Areas (23116516) from The Ministry of Education, Culture, Sports, Science and Technology of Japan, and by grants from Japan Foundation for Applied Enzymology, Takeda Science Foundation, The Naito Foundation, and The Research Foundation for Pharmaceutical Sciences (KF). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.