title: Nitric oxide generation affects pro- and anti-angiogenic growth factor expression in primary human trophoblast.
abstract: OBJECTIVE
  Preeclampsia is associated with reduced trophoblast placenta growth factor (PGF) expression, elevated soluble fms-like tyrosine kinase-1 (sFlt-1) and decreased bioactivity of nitric oxide (NO). Elevated sFlt-1 reduces bio-availability of PGF and vascular endothelial growth factor (VEGF) leading to maternal endothelial dysfunction. Although NO can regulate gene expression, its ability to regulate trophoblast expression of angiogenic growth factors is not known.
  
  
  METHODS
  Human primary term trophoblast and JEG-3 choriocarcinoma cells were cultured under 21%O(2) or 1%O(2) conditions in the presence or absence of NO donor (SNP) or inhibitor (L-NAME). Effects on PGF, VEGF and Flt-1 isoform mRNA expression were determined by quantitative real-time PCR. Changes in expression of soluble protein isoforms of FLT-1 was monitored by ELISA.
  
  
  RESULTS
  Hypoxia decreased PGF mRNA but increased VEGF, sFlt-1 and Flt-1 mRNA expression in trophoblast. Generation of NO in trophoblast under 1%O(2) culture conditions significantly reversed sFlt-1 mRNA and protein expression, independent of mFlt-1. Conversely NO generation in hypoxic trophoblast increased VEGF and PGF mRNA expression.
  
  
  CONCLUSIONS
  NO production in primary human trophoblast cultures had divergent effects on pro-angiogenic (PGF, VEGF) versus anti-angiogenic (sFlt-1) mRNA expression, resulting in an enhanced pro-angiogenic gene expression environment in vitro.
authors.affiliation: Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL, USA.
authors.forename: K A
authors.initials: KA
authors.lastname: Groesch
authors.forename: R J
authors.initials: RJ
authors.lastname: Torry
authors.forename: A C
authors.initials: AC
authors.lastname: Wilber
authors.forename: R
authors.initials: R
authors.lastname: Abrams
authors.forename: A
authors.initials: A
authors.lastname: Bieniarz
authors.forename: L J
authors.initials: LJ
authors.lastname: Guilbert
authors.forename: D S
authors.initials: DS
authors.lastname: Torry
mesh_terms: D002478:Cells, Cultured; D005260:Female; D006801:Humans; D000860:Hypoxia / Q000378:metabolism*; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology*; D009569:Nitric Oxide / Q000096:biosynthesis*; D009599:Nitroprusside / Q000378:metabolism; D000072483:Placenta Growth Factor; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000378:metabolism; D014327:Trophoblasts / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000096:biosynthesis*
chemical_list: C000608036:PGF protein, human; D011257:Pregnancy Proteins; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D000072483:Placenta Growth Factor; D009599:Nitroprusside; D009569:Nitric Oxide; C501162:FLT1 protein, human; D040281:Vascular Endothelial Growth Factor Receptor-1; D019331:NG-Nitroarginine Methyl Ester