Jiang Zhong L, Zhu Xuping, Diamond Michael P, Abu-Soud Husam M, Saed Ghassan M
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.
Fertil Steril. 2008 Sep;90(3):769-74. doi: 10.1016/j.fertnstert.2007.07.1313. Epub 2008 Apr 28.
To determine the expression of nitric oxide synthases (NOSs) and their modulation by hypoxia in human peritoneal (NF) and adhesion fibroblasts (ADF).
Prospective experimental study.
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).
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.
确定一氧化氮合酶（NOSs）在人腹膜成纤维细胞（NF）和粘连成纤维细胞（ADF）中的表达及其受缺氧的调节情况。
前瞻性实验研究。
大学医学中心。
腹膜和粘连组织的成纤维细胞。
成纤维细胞中的缺氧和沉默诱导型一氧化氮合酶（iNOS）基因表达。
我们使用逆转录聚合酶链反应来定量NOS亚型的信使核糖核酸（mRNA）水平。采用格里斯试剂法测量一氧化氮水平。
神经元型一氧化氮合酶（nNOS）和内皮型一氧化氮合酶（eNOS）的mRNA拷贝数/微克RNA在NF中分别为6.6×10³（均值）、5.7×10³（均值），在ADF中分别为7.0×10³（均值）、6.1×10³（均值）。iNOS的mRNA拷贝数/微克RNA在NF中为31.3×10³，在ADF中为33.0×10³。缺氧使NF中iNOS的mRNA拷贝数/微克RNA从31.3×10³增加到61.3×10³，使ADF中从33.0×10³增加到63.9×10³，而NF和ADF中nNOS和eNOS的mRNA水平无变化。ADF中的一氧化氮水平（0.94微摩尔/升）低于NF（1.97微摩尔/升）。沉默iNOS可降低NF中的一氧化氮水平（从1.97微摩尔/升降至0.41微摩尔/升）以及ADF中的一氧化氮水平（从0.94微摩尔/升降至0.27微摩尔/升）。
一氧化氮合酶在NF和ADF中差异表达，其中iNOS表达最多且是一氧化氮的主要来源。缺氧被证明可改变NF和ADF中一氧化氮合酶和一氧化氮的表达。
