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

人正常腹膜和粘连组织中分离的成纤维细胞中一氧化氮合酶亚型的表达

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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/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).

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