Gender and transcriptional regulation of NO synthase and ET-1 in porcine aortic endothelial cells.
Gender and transcriptional regulation of NO synthase and ET-1 in porcine aortic endothelial cells.
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DOI：
10.1152/ajpheart.1997.273.4.h1962
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发表时间：
1997-10
期刊：
影响因子：
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通讯作者：
Xiaofang Wang;D. A. Barber;D. Lewis;C. McGregor;G. Sieck;L. Fitzpatrick;V. Miller
中科院分区：
文献类型：
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作者：
Xiaofang Wang;D. A. Barber;D. Lewis;C. McGregor;G. Sieck;L. Fitzpatrick;V. Miller
实验旨在确定性类固醇激素的正常波动是否会改变内皮型一氧化氮合酶（NOS）和内皮素原-1（prepro-ET-1）的基因转录。从成年、性腺完整的雄性和雌性或卵巢切除的约克郡猪中取出主动脉内皮细胞。制备内皮细胞用于北方印迹分析、Western印迹分析或酶活性。分别用化学发光法和放射免疫法测定血浆一氧化氮产物（NOx）和内皮素-1（ET-1）。北方印迹分析鉴定了对应于内皮NOS和前ET-1原的单一条带。定量的印迹显示，增加的mRNA表达的内皮NOS和prepro-ET-1的卵巢切除猪相比，性腺完整的男性和女性的猪。Westernblot分析显示各组间内皮细胞NOS蛋白表达量无差异。相反，血浆中的NOx浓度显着降低卵巢切除猪，有没有差异，无论是在血浆中的ET-1的浓度或提取物从冠状动脉。提示卵巢激素可能在转录水平调控内皮NOS和ET-1前体的表达。此外，卵巢激素可能在转录后水平调节这些内皮衍生因子的产生。
Experiments were designed to determine whether normal fluctuations in sex steroid hormones alter gene transcription for endothelial nitric oxide synthase (NOS) and preproendothelin-1 (prepro-ET-1). Aortic endothelial cells were removed from adult, gonadally intact male and female or ovariectomized Yorkshire pigs. Endothelial cells were prepared for Northern blot analysis, Western blot analysis or enzyme activity. Nitric oxide products (NOx) and endothelin-1 (ET-1) in plasma were measured by chemiluminescence and radioimmunoassay, respectively. Northern blot analysis identified single bands corresponding to endothelial NOS and prepro-ET-1. Quantification of the blots showed an increase in expression of mRNA for both endothelial NOS and prepro-ET-1 in ovariectomized pigs compared with gonadally intact male and female pigs. There were no differences in amount of endothelial NOS protein identified by Western blot analysis among groups. On the contrary, plasma concentrations of NOx were significantly decreased in ovariectomized pigs, and there were no differences either in the concentrations of ET-1 in the plasma or extracts from the coronary arteries. These results suggest that expression of endothelial NOS and prepro-ET-1 may be regulated at transcriptional level by ovarian hormones. In addition, the ovarian hormones may regulate production of these endothelium-derived factors at the posttranscriptional level.