Nitric oxide: an autocrine regulator of human granulosa-luteal cell steroidogenesis.
Nitric oxide: an autocrine regulator of human granulosa-luteal cell steroidogenesis.
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DOI：
10.1210/endo.135.5.7525252
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发表时间：
1994-11
期刊：
影响因子：
4.8
通讯作者：
B J Van Voorhis;M. Dunn;G D Snyder;C. P. Weiner
中科院分区：
文献类型：
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作者：
B J Van Voorhis;M. Dunn;G D Snyder;C. P. Weiner
我们研究了一氧化氮（NO）合酶的存在下，卵巢卵泡细胞从妇女接受体外受精程序。逆转录聚合酶链反应扩增内皮细胞NO合成酶mRNA。NO合酶定位于颗粒黄体细胞免疫细胞化学，使用单克隆抗体。通过测量L-精氨酸转化为瓜氨酸来证实卵巢卵泡细胞NO合酶的活性。为了研究NO对颗粒黄体细胞类固醇生成的影响，将NO合酶抑制剂和NO供体添加到细胞培养物中。NO合成酶的选择性抑制剂NG-单甲基-L-精氨酸和N-硝基-精氨酸酶甲酯可显著增加颗粒黄体细胞的雌二醇分泌。S-亚硝基-L-乙酰青霉胺（S-NAP）和S-亚硝基谷胱甘肽，NO供体，引起剂量依赖性降低雌二醇和孕酮分泌。由S-NAP的减少被血红蛋白，它结合游离NO逆转。虽然S-NAP增加了颗粒黄体细胞中cGMP的浓度，cGMP类似物对细胞培养物中的类固醇生成没有影响。S-NAP和天然NO在溶液中降低细胞和微粒体芳香化酶活性。我们的结论是，NO合酶是存在于人类颗粒黄体细胞和NO抑制雌二醇分泌的cGMP通过直接抑制芳香化酶独立。
We investigated the presence of nitric oxide (NO) synthase in ovarian follicular cells obtained from women undergoing in vitro fertilization procedures. Endothelial NO synthase messenger RNA was demonstrated by polymerase chain reaction amplification of reverse transcribed RNA. NO synthase was localized to granulosa-luteal cells by immunocytochemistry, using a monoclonal antibody. Ovarian follicular cell NO synthase enzyme activity was confirmed by measuring the conversion of L-arginine to citrulline. To investigate the effect of NO on granulosa-luteal cell steroidogenesis, NO synthase inhibitors and NO donors were added to cell cultures. NG-Monomethyl-L-arginine and N-nitro-arginase methyl ester, selective inhibitors of NO synthase, significantly increased estradiol secretion by granulosa-luteal cells. S-Nitroso-L-acetyl penicillamine (S-NAP) and S-nitroso glutathione, NO donors, caused a dose-dependent decrease in both estradiol and progesterone secretion. The decrease by S-NAP was reversed by hemoglobin, which binds free NO. Although S-NAP increased the concentration of cGMP in granulosa-luteal cells, cGMP analogs had no effect on steroidogenesis in cell cultures. S-NAP and native NO in solution decreased cellular and microsomal aromatase activities. We conclude that NO synthase is present in human granulosa-luteal cells and that NO inhibits estradiol secretion independent of cGMP by directly inhibiting aromatase.