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通过逆转录聚合酶链反应(RT-PCR)对卵丘细胞中类固醇硫酸酯酶进行定位和基因表达分析及其与体外受精期间观察到的血清卵泡刺激素(FSH)水平的关系。

Localization and gene expression of steroid sulfatase by RT-PCR in cumulus cells and relationship to serum FSH levels observed during in vitro fertilization.

作者信息

Otsuka Yukiko, Yanaihara Atsushi, Iwasaki Shinji, Hasegawa Junichi, Yanaihara Takumi, Okai Takashi

机构信息

Department of Obstetrics and Gynecology Showa University School of Medicine 1-5-8, Hatanodai, Shinagawa-ku, Tokyo, Japan.

出版信息

J Exp Clin Assist Reprod. 2005 Apr 11;2(1):6. doi: 10.1186/1743-1050-2-6.

Abstract

BACKGROUND

The purpose of this study was to localize the expression of steroid sulfatase (STS) in cumulus cells and to determine the relationship between STS mRNA expression and the serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol and progesterone. METHODS: The subject group included 49 women (29 to 44 years old) for whom in vitro fertilization treatment was indicated. All subjects gave informed consent. One hundred fourteen samples of cumulus-oocyte complex (COC) were obtained under microscopic observation. Part of the COC was stained by STS antibody. RNA was extracted by phenol-chloroform method and real-time PCR was performed. Serum of each patient was collected and was measured by ELISA. RESULTS: Some of the cumulus samples were stained by STS antibody. The expression of STS mRNA in all samples was confirmed by quantitative RT-PCR. Although there was no significant correlation between the level of STS mRNA and the serum levels of estradiol, progesterone and LH, there was a statistically significant negative correlation between the level of STS mRNA expression and the serum level of FSH (n = 105, p = 0.018, r = -0.22). CONCLUSION: These results have demonstrated for the first time the expression of STS in cumulus cells by immunohistological stainings and real-time RT-PCR. STS expression in cumulus cells may be related to the control of the local steroidal environment in the oocyte. Serum FSH may control STS mRNA expression from the results of RT-PCR, although the correlation was low.

摘要

背景

本研究旨在定位类固醇硫酸酯酶(STS)在卵丘细胞中的表达,并确定STS mRNA表达与血清促卵泡激素(FSH)、促黄体生成素(LH)、雌二醇和孕酮水平之间的关系。方法:研究对象包括49名(29至44岁)有体外受精治疗指征的女性。所有受试者均签署知情同意书。在显微镜观察下获取114个卵丘-卵母细胞复合体(COC)样本。部分COC用STS抗体染色。采用酚-氯仿法提取RNA并进行实时PCR。收集每位患者的血清,用ELISA法检测。结果:部分卵丘样本被STS抗体染色。通过定量RT-PCR证实了所有样本中STS mRNA的表达。虽然STS mRNA水平与雌二醇、孕酮和LH的血清水平之间无显著相关性,但STS mRNA表达水平与FSH血清水平之间存在统计学上的显著负相关(n = 105,p = 0.018,r = -0.22)。结论:这些结果首次通过免疫组织化学染色和实时RT-PCR证明了STS在卵丘细胞中的表达。卵丘细胞中STS的表达可能与卵母细胞局部甾体环境的调控有关。尽管相关性较低,但RT-PCR结果表明血清FSH可能控制STS mRNA的表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a14c/1082910/c82300b57b98/1743-1050-2-6-1.jpg

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