Nitric oxide synthase isoforms expression in fibroblasts isolated from human normal peritoneum and adhesion tissues

Zhong L. Jiang, Xuping Zhu, Michael Peter Diamond, Husam M. Abu-Soud, Ghassan M. Saed

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Objective: To determine the expression of nitric oxide synthases (NOSs) and their modulation by hypoxia in human peritoneal (NF) and adhesion fibroblasts (ADF). Design: Prospective experimental study. Setting: University medical center. Patient(s): Fibroblasts from peritoneum and adhesion tissues. Intervention(s): Hypoxia and silencing inducible NOS (iNOS) gene expression in fibroblasts. Main Outcome Measure(s): We used reverse-transcriptase polymerase chain reaction to quantify messenger RNA (mRNA) levels of NOS isoforms. Griess assay was used to measure NO levels. Result(s): The mRNA copies/μg RNA of neuronal NOS (nNOS) and endothelial NOS (eNOS) were 6.6 × 103 in NF, 5.7 × 103 in ADF and 7.0 × 103 in NF, 6.1 × 103 in ADF, respectively. The mRNA copies/μg RNA of iNOS were 31.3 × 103 in NF and 33.0 × 103 in ADF. Hypoxia increased iNOS mRNA copies/μg RNA from 31.3 × 103 to 61.3 × 103 in NF and from 33.0 × 103 to 63.9 × 103 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 μmol/L) than NF (1.97 μmol/L). Silencing iNOS decreased NO levels in NF (from 1.97 μmol/L to 0.41 μmol/L) and in ADF (from 0.94 μmol/L to 0.27 μmol/L). Conclusion(s): 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.

Original languageEnglish (US)
Pages (from-to)769-774
Number of pages6
JournalFertility and sterility
Volume90
Issue number3
DOIs
StatePublished - Sep 1 2008
Externally publishedYes

Fingerprint

Tissue Adhesions
Peritoneum
Nitric Oxide Synthase
Protein Isoforms
Fibroblasts
Messenger RNA
RNA
Nitric Oxide Synthase Type I
Nitric Oxide Synthase Type III
Nitric Oxide Synthase Type II
Reverse Transcriptase Polymerase Chain Reaction

Keywords

  • Adhesions
  • RT-PCR
  • fibroblasts
  • hypoxia
  • nitric oxide
  • nitric oxide synthase
  • peritoneum
  • siRNA

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology

Cite this

Nitric oxide synthase isoforms expression in fibroblasts isolated from human normal peritoneum and adhesion tissues. / Jiang, Zhong L.; Zhu, Xuping; Diamond, Michael Peter; Abu-Soud, Husam M.; Saed, Ghassan M.

In: Fertility and sterility, Vol. 90, No. 3, 01.09.2008, p. 769-774.

Research output: Contribution to journalArticle

Jiang, Zhong L. ; Zhu, Xuping ; Diamond, Michael Peter ; Abu-Soud, Husam M. ; Saed, Ghassan M. / Nitric oxide synthase isoforms expression in fibroblasts isolated from human normal peritoneum and adhesion tissues. In: Fertility and sterility. 2008 ; Vol. 90, No. 3. pp. 769-774.
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T1 - Nitric oxide synthase isoforms expression in fibroblasts isolated from human normal peritoneum and adhesion tissues

AU - Jiang, Zhong L.

AU - Zhu, Xuping

AU - Diamond, Michael Peter

AU - Abu-Soud, Husam M.

AU - Saed, Ghassan M.

PY - 2008/9/1

Y1 - 2008/9/1

N2 - Objective: To determine the expression of nitric oxide synthases (NOSs) and their modulation by hypoxia in human peritoneal (NF) and adhesion fibroblasts (ADF). Design: Prospective experimental study. Setting: University medical center. Patient(s): Fibroblasts from peritoneum and adhesion tissues. Intervention(s): Hypoxia and silencing inducible NOS (iNOS) gene expression in fibroblasts. Main Outcome Measure(s): We used reverse-transcriptase polymerase chain reaction to quantify messenger RNA (mRNA) levels of NOS isoforms. Griess assay was used to measure NO levels. Result(s): The mRNA copies/μg RNA of neuronal NOS (nNOS) and endothelial NOS (eNOS) were 6.6 × 103 in NF, 5.7 × 103 in ADF and 7.0 × 103 in NF, 6.1 × 103 in ADF, respectively. The mRNA copies/μg RNA of iNOS were 31.3 × 103 in NF and 33.0 × 103 in ADF. Hypoxia increased iNOS mRNA copies/μg RNA from 31.3 × 103 to 61.3 × 103 in NF and from 33.0 × 103 to 63.9 × 103 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 μmol/L) than NF (1.97 μmol/L). Silencing iNOS decreased NO levels in NF (from 1.97 μmol/L to 0.41 μmol/L) and in ADF (from 0.94 μmol/L to 0.27 μmol/L). Conclusion(s): 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.

AB - Objective: To determine the expression of nitric oxide synthases (NOSs) and their modulation by hypoxia in human peritoneal (NF) and adhesion fibroblasts (ADF). Design: Prospective experimental study. Setting: University medical center. Patient(s): Fibroblasts from peritoneum and adhesion tissues. Intervention(s): Hypoxia and silencing inducible NOS (iNOS) gene expression in fibroblasts. Main Outcome Measure(s): We used reverse-transcriptase polymerase chain reaction to quantify messenger RNA (mRNA) levels of NOS isoforms. Griess assay was used to measure NO levels. Result(s): The mRNA copies/μg RNA of neuronal NOS (nNOS) and endothelial NOS (eNOS) were 6.6 × 103 in NF, 5.7 × 103 in ADF and 7.0 × 103 in NF, 6.1 × 103 in ADF, respectively. The mRNA copies/μg RNA of iNOS were 31.3 × 103 in NF and 33.0 × 103 in ADF. Hypoxia increased iNOS mRNA copies/μg RNA from 31.3 × 103 to 61.3 × 103 in NF and from 33.0 × 103 to 63.9 × 103 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 μmol/L) than NF (1.97 μmol/L). Silencing iNOS decreased NO levels in NF (from 1.97 μmol/L to 0.41 μmol/L) and in ADF (from 0.94 μmol/L to 0.27 μmol/L). Conclusion(s): 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.

KW - Adhesions

KW - RT-PCR

KW - fibroblasts

KW - hypoxia

KW - nitric oxide

KW - nitric oxide synthase

KW - peritoneum

KW - siRNA

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