Abstract
To study the modulation of the inducible nitric oxide synthase/nitric oxide (iNOS/NO) expression system in fibroblasts isolated from human peritoneum and adhesion tissues by hypoxia. Prospective experimental study. University medical center. Cultures of fibroblasts from both peritoneum and adhesion tissues of five patients. Hypoxia treatment of the primary cultured fibroblasts. We used Western and Northern blots to determine whether iNOS mRNA and its protein were present in peritoneal and adhesion fibroblasts and whether this expression is modulated by hypoxia. Multiplex reverse transcription polymerase chain reaction (RT-PCR) technique was used to quantify type I collagen mRNA in response to N G-nitro-L-arginine methyl ester (L-NAME). A terminal deoxynucleotidyl transferase mediated dUTP nick-end-labeling (TUNEL) assay was used to quantify apoptosis in response to NO donor S-nitro-N-acetyl- penicillamine (SNAP) treatment. A Griess assay was used to measure NO levels. Peritoneal fibroblasts have significantly higher NO levels than adhesion fibroblasts. Hypoxia decreased NO in peritoneal fibroblasts to levels observed for adhesion fibroblasts. In addition, hypoxia increased both mRNA and protein levels of the iNOS gene in peritoneal and adhesion fibroblasts. Augmentation of NO by SNAP treatment increased apoptosis in adhesion fibroblasts. In contrast, SNAP had no effect on apoptosis of peritoneal fibroblasts. Inhibition of NO by L-NAME treatment increased type I collagen mRNA levels in peritoneal fibroblasts. Our findings confirm that adhesion fibroblasts produce less NO than normal peritoneal fibroblasts; NO may be the mechanism responsible for the creation and persistence of the adhesion phenotype.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1198-1205 |
| Number of pages | 8 |
| Journal | Fertility and Sterility |
| Volume | 82 |
| Issue number | SUPPL. 3 |
| DOIs | |
| State | Published - Oct 1 2004 |
| Externally published | Yes |
Fingerprint
Keywords
- adhesion
- Fibroblasts
- hypoxia
- nitric oxide
- peritoneum
ASJC Scopus subject areas
- Obstetrics and Gynecology
Cite this
Role of nitric oxide in apoptosis of human peritoneal and adhesion fibroblasts after hypoxia. / Saed, Ghassan M.; Abu-Soud, Husam M.; Diamond, Michael Peter.
In: Fertility and Sterility, Vol. 82, No. SUPPL. 3, 01.10.2004, p. 1198-1205.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Role of nitric oxide in apoptosis of human peritoneal and adhesion fibroblasts after hypoxia
AU - Saed, Ghassan M.
AU - Abu-Soud, Husam M.
AU - Diamond, Michael Peter
PY - 2004/10/1
Y1 - 2004/10/1
N2 - To study the modulation of the inducible nitric oxide synthase/nitric oxide (iNOS/NO) expression system in fibroblasts isolated from human peritoneum and adhesion tissues by hypoxia. Prospective experimental study. University medical center. Cultures of fibroblasts from both peritoneum and adhesion tissues of five patients. Hypoxia treatment of the primary cultured fibroblasts. We used Western and Northern blots to determine whether iNOS mRNA and its protein were present in peritoneal and adhesion fibroblasts and whether this expression is modulated by hypoxia. Multiplex reverse transcription polymerase chain reaction (RT-PCR) technique was used to quantify type I collagen mRNA in response to N G-nitro-L-arginine methyl ester (L-NAME). A terminal deoxynucleotidyl transferase mediated dUTP nick-end-labeling (TUNEL) assay was used to quantify apoptosis in response to NO donor S-nitro-N-acetyl- penicillamine (SNAP) treatment. A Griess assay was used to measure NO levels. Peritoneal fibroblasts have significantly higher NO levels than adhesion fibroblasts. Hypoxia decreased NO in peritoneal fibroblasts to levels observed for adhesion fibroblasts. In addition, hypoxia increased both mRNA and protein levels of the iNOS gene in peritoneal and adhesion fibroblasts. Augmentation of NO by SNAP treatment increased apoptosis in adhesion fibroblasts. In contrast, SNAP had no effect on apoptosis of peritoneal fibroblasts. Inhibition of NO by L-NAME treatment increased type I collagen mRNA levels in peritoneal fibroblasts. Our findings confirm that adhesion fibroblasts produce less NO than normal peritoneal fibroblasts; NO may be the mechanism responsible for the creation and persistence of the adhesion phenotype.
AB - To study the modulation of the inducible nitric oxide synthase/nitric oxide (iNOS/NO) expression system in fibroblasts isolated from human peritoneum and adhesion tissues by hypoxia. Prospective experimental study. University medical center. Cultures of fibroblasts from both peritoneum and adhesion tissues of five patients. Hypoxia treatment of the primary cultured fibroblasts. We used Western and Northern blots to determine whether iNOS mRNA and its protein were present in peritoneal and adhesion fibroblasts and whether this expression is modulated by hypoxia. Multiplex reverse transcription polymerase chain reaction (RT-PCR) technique was used to quantify type I collagen mRNA in response to N G-nitro-L-arginine methyl ester (L-NAME). A terminal deoxynucleotidyl transferase mediated dUTP nick-end-labeling (TUNEL) assay was used to quantify apoptosis in response to NO donor S-nitro-N-acetyl- penicillamine (SNAP) treatment. A Griess assay was used to measure NO levels. Peritoneal fibroblasts have significantly higher NO levels than adhesion fibroblasts. Hypoxia decreased NO in peritoneal fibroblasts to levels observed for adhesion fibroblasts. In addition, hypoxia increased both mRNA and protein levels of the iNOS gene in peritoneal and adhesion fibroblasts. Augmentation of NO by SNAP treatment increased apoptosis in adhesion fibroblasts. In contrast, SNAP had no effect on apoptosis of peritoneal fibroblasts. Inhibition of NO by L-NAME treatment increased type I collagen mRNA levels in peritoneal fibroblasts. Our findings confirm that adhesion fibroblasts produce less NO than normal peritoneal fibroblasts; NO may be the mechanism responsible for the creation and persistence of the adhesion phenotype.
KW - adhesion
KW - Fibroblasts
KW - hypoxia
KW - nitric oxide
KW - peritoneum
UR - http://www.scopus.com/inward/record.url?scp=5044221808&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=5044221808&partnerID=8YFLogxK
U2 - 10.1016/j.fertnstert.2004.04.034
DO - 10.1016/j.fertnstert.2004.04.034
M3 - Article
C2 - 15474096
AN - SCOPUS:5044221808
VL - 82
SP - 1198
EP - 1205
JO - Fertility and Sterility
JF - Fertility and Sterility
SN - 0015-0282
IS - SUPPL. 3
ER -