国际生殖健康/计划生育杂志

国际生殖健康/计划生育 ›› 2011, Vol. 30 ›› Issue (2): 88-94.

• 论著 • 上一篇    下一篇

核受体NR4A1在卵巢卵泡膜细胞内调节雄激素的生成

李 梅 刁飞扬 薛 凯 凌 静 丁 卫 蔡令波 崔毓桂 刘嘉茵   

  1. 210029 南京医科大学第一附属医院生殖医学科 江苏省生殖医学重点实验室
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-03-15 发布日期:2011-03-15
  • 通讯作者: 刁飞扬,刘嘉茵

The Orphan Nuclear Receptor NR4A1 Regulates Androgen Production in Ovarian Theca Cells

LI Mei， DIAO Fei-yang， XUE Kai， LING Jing， DING Wei， CAI Ling-bo， CUI Yu-gui， LIU Jia-yin   

  1. Jiangsu Province Key Laboratory of Reproductive Medicine Nanjing Medical University， Center of Clinical Reproductive Medicine， the First Affiliated Hospital，Nanjing Medical University，Nanjing 210029， China
  • Received:1900-01-01 Revised:1900-01-01 Published:2011-03-15 Online:2011-03-15
  • Contact: DIAO Fei-yang，LIU Jia-yin

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摘要： 目的：孤儿核受体NR4A1是核受体超家族中的一员，其广泛分布于不同的细胞类型并参与多种生物学过程的调控。探讨在卵泡膜细胞中NR4A1对甾体激素生成酶的转录及雄激素生成的影响。方法：构建小鼠NR4A1重组腺病毒载体AdCMV?鄄NR4A1及AdH1?鄄NR4A1，重组病毒感染卵泡24 h后，蛋白质印迹（Western Blot）验证NR4A1在卵泡膜细胞中的超表达及干涉效果。实时定量反转录聚合酶链反应（Real?鄄time RT?鄄PCR）检测Nr4a1超表达及干涉后对雄激素生成酶（Star，Cyp11a1，Cyp17a1，Hsd3b2）的转录调节，放射免疫法测定培养液中睾酮的浓度。结果：AdCMV?鄄NR4A1感染卵泡膜细胞后NR4A1蛋白的表达水平显著升高。NR4A1在卵泡膜细胞内超表达后明显促进Star，Cyp11a1，Cyp17a1，Hsd3b2的转录，增加睾酮合成。相反，AdH1-NR4A1感染卵泡膜细胞后，NR4A1蛋白的表达水平显著降低。NR4A1干涉后明显抑制Star，Cyp11a1，Cyp17a1，Hsd3b2的表达，降低睾酮的合成。此外，在卵泡培养液中加入cAMP/PKA通路激动剂——福斯高林后，Nr4a1的表达迅速增加，随后Star，Cyp11a1，Cyp17a1，Hsd3b2的表达增加。结论：核受体NR4A1在卵泡膜细胞内促进雄激素生成酶的转录，进而调节雄激素生成。该研究为进一步探讨NR4A1在多囊卵巢综合征高雄激素血症等病理生理改变中的分子调控机制奠定了基础。

关键词: 核受体NR4A1, 多囊卵巢综合征, 高雄激素血症

Abstract: Objective： Orphan nuclear receptor NR4A1， a member of the nuclear receptor superfamily，is widely expressed in different cell types and mediates diverse biological processes. In the present study，our main purpose was to explore the function of NR4A1 in the transcript regulation of steroidogenic enzyme genes responsible for ovarian theca cell steroidogenesis. Methods： We constructed recombinant adenovirus AdCMV?-NR4A1 and AdH1-NR4A1 to enhance or knockdown the expression of NR4A1 in theca cells，respectively. The expression patterns of Cyp11a1， Cyp17a1 and Hsd3b2 were subsequently analyzed by real?-time RT-PCR. Moreover，concentrations of testosterone in the spent medium were measured by radioimmunoassay. Results：Our results show that overexpression of NR4A1 in theca cells stimulates the expression of Star， Cyp11a1， Cyp17a1 and Hsd3b2， leading to increased testosterone production. Conversely，knockdown of the endogenous NR4A1 exhibits a significant decrease in Cyp11a1， Cyp17a1 and Hsd3b2 expression and testosterone production. Forskolin（FSK）， an activator of cAMP/PKA pathway，was applied to the cultured follicles. FSK rapidly increases the Nr4a1 mRNA levels followed by an increase in Cyp11a1， Cyp17a1 and Hsd3b2. Conclusions：Collectively， our results outline a previously unrecognized role for NR4A1 in the transcriptional regulation of Cyp11a1， Cyp17a1 and Hsd3b2 in ovarian theca cells. Modulation of these steroidogenic enzymes by NR4A1 could influence the capacity of the ovarian theca cells to produce androgen. This study provide basis for us to further explore the mechanism of NR4A1 on the pathophysiology development of polycystic ovary syndrome hyperandrogenism at molecular level.

Key words: Nuclear receptor NR4A1, Polycystic ovary syndrome, Hyperandrogenism