17beta-estradiol up-regulates prostacyclin production in cultured human uterine myometrial cells via augmentation of both cyclooxygenase-1 and prostacyclin synthase expression

J Soc Gynecol Investig. 2004 Oct;11(7):457-64. doi: 10.1016/j.jsgi.2004.04.002.

Abstract

Objective: To investigate whether 17beta-estradiol elevates prostacyclin (PGI(2)) production in human myometrial cells in the middle of gestation.

Methods: The concentration of 6-keto-PGF(1alpha), a stable metabolite of PGI(2), in the culture medium was assessed using enzyme-linked immunosorbent assay (ELISA). Western blot analysis and quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) using TaqMan (Applied Biosystems, Foster City, CA) technology were performed to evaluate the expression of cytosolic phopholipase A(2) (cPLA(2)), cyclooxygenase-1 (COX-1), COX-2, and prostacyclin synthase (PGIS) in cultured human myometrial cells prepared from second trimester pregnant women (n = 3) after stimulation with 17beta-estradiol.

Results: Treatment with 17beta-estradiol (4-400 nM) dose-dependently elevated PGI(2) secretion from cultured human myometrial cells. Western blot analysis detected cPLA(2) and COX-1 and PGIS protein expression in the cultured human myometrial cells; however, COX-2 protein expression was below the detection sensitivity. Stimulation with 40-nM 17beta-estradiol significantly up-regulated protein and mRNA expression of both COX-1 and PGIS.

Conclusion: 17beta-Estradiol from placenta may contribute to the augmentation of PGI(2) production in the human myometrium in the middle of gestation via up-regulation of both COX-1 and PGIS expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 6-Ketoprostaglandin F1 alpha / metabolism
  • Blotting, Western
  • Cells, Cultured
  • Cyclooxygenase 1
  • Cyclooxygenase 2
  • Cytochrome P-450 Enzyme System / analysis
  • Cytochrome P-450 Enzyme System / genetics*
  • Epoprostenol / biosynthesis*
  • Estradiol / pharmacology*
  • Female
  • Gene Expression / drug effects*
  • Gestational Age
  • Humans
  • Intramolecular Oxidoreductases / analysis
  • Intramolecular Oxidoreductases / genetics*
  • Membrane Proteins
  • Myometrium / drug effects*
  • Myometrium / enzymology
  • Myometrium / metabolism
  • Phospholipases A / genetics
  • Pregnancy
  • Prostaglandin-Endoperoxide Synthases / analysis
  • Prostaglandin-Endoperoxide Synthases / genetics*
  • RNA, Messenger / analysis
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Membrane Proteins
  • RNA, Messenger
  • Estradiol
  • 6-Ketoprostaglandin F1 alpha
  • Cytochrome P-450 Enzyme System
  • Epoprostenol
  • Cyclooxygenase 1
  • Cyclooxygenase 2
  • PTGS1 protein, human
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases
  • Phospholipases A
  • Intramolecular Oxidoreductases
  • prostacyclin synthetase