title: Nitric oxide synthase isoforms expression in fibroblasts isolated from human normal peritoneum and adhesion tissues.
abstract: OBJECTIVE
  To determine the expression of nitric oxide synthases (NOSs) and their modulation by hypoxia in human peritoneal (NF) and adhesion fibroblasts (ADF).
  
  
  METHODS
  Prospective experimental study.
  
  
  METHODS
  University medical center.
  
  
  METHODS
  Fibroblasts from peritoneum and adhesion tissues.
  
  
  METHODS
  Hypoxia and silencing inducible NOS (iNOS) gene expression in fibroblasts.
  
  
  METHODS
  We used reverse-transcriptase polymerase chain reaction to quantify messenger RNA (mRNA) levels of NOS isoforms. Griess assay was used to measure NO levels.
  
  
  RESULTS
  The mRNA copies/mug RNA of neuronal NOS (nNOS) and endothelial NOS (eNOS) were 6.6 x 10(3) in NF, 5.7 x 10(3) in ADF and 7.0 x 10(3) in NF, 6.1 x 10(3) in ADF, respectively. The mRNA copies/mug RNA of iNOS were 31.3 x 10(3) in NF and 33.0 x 10(3) in ADF. Hypoxia increased iNOS mRNA copies/mug RNA from 31.3 x 10(3) to 61.3 x 10(3) in NF and from 33.0 x 10(3) to 63.9 x 10(3) in ADF, whereas there were no changes in mRNA levels of nNOS and eNOS in NF and ADF. Nitric oxide levels were lower in ADF (0.94 micromol/L) than NF (1.97 micromol/L). Silencing iNOS decreased NO levels in NF (from 1.97 micromol/L to 0.41 micromol/L) and in ADF (from 0.94 micromol/L to 0.27 micromol/L).
  
  
  CONCLUSIONS
  Nitric oxide synthases are differentially expressed in NF and ADF, with iNOS being the most expressed and the main source of NO. Hypoxia was shown to alter the expression of NOSs and NO in NF and ADF.
authors.affiliation: Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, The C S Mott Center for Human Growth and Development, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
authors.forename: Zhong L
authors.initials: ZL
authors.lastname: Jiang
authors.forename: Xuping
authors.initials: X
authors.lastname: Zhu
authors.forename: Michael P
authors.initials: MP
authors.lastname: Diamond
authors.forename: Husam M
authors.initials: HM
authors.lastname: Abu-Soud
authors.forename: Ghassan M
authors.initials: GM
authors.lastname: Saed
mesh_terms: D002448:Cell Adhesion; D015687:Cell Hypoxia / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D010537:Peritoneum / Q000201:enzymology*; D014018:Tissue Distribution
chemical_list: D007527:Isoenzymes; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase