Nitric oxide synthase isoforms expression in fibroblasts isolated from human normal peritoneum and adhesion tissues.
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.
Jiang
Zhong L
ZL
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.
Zhu
Xuping
X
Diamond
Michael P
MP
Abu-Soud
Husam M
HM
Saed
Ghassan M
GM
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
D007527:Isoenzymes; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase