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山东大学学报（医学版） ›› 2016, Vol. 54 ›› Issue (5): 50-55.doi: 10.6040/j.issn.1671-7554.0.2015.437

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雌激素对卵巢颗粒细胞雌激素受体-β和转录因子叉头蛋白3表达的影响

马会明1,3,张永芳1,2,王蒙蒙1,李昕1,王永峰1,2,田洪成1,胡蓉1,2,王燕蓉1,3,裴秀英1,徐仙1,2   

  1. 1.宁夏医科大学生育力保持教育部重点实验室&宁夏回族自治区生殖与遗传重点实验室, 宁夏 银川 750004;2.宁夏医科大学总医院生殖医学中心, 宁夏 银川 750004;3.宁夏医科大学人体解剖与组织胚胎学系组织胚胎学教研室, 宁夏 银川 750004
  • 收稿日期:2015-04-30 出版日期:2016-05-16 发布日期:2016-05-16
  • 通讯作者: 裴秀英. E-mail:peixiuying@163.com徐仙. E-mail:xuxjournal@163.com E-mail:peixiuying@163.com
  • 基金资助:
    国家自然科学基金(31200882)

Effect of estrogen on expression of estrogen receptor-β and forkhead transcription factor 3 in human granulosa cells

MA Huiming1,3, ZHANG Yongfang1,2, WANG Mengmeng1, LI Xin1, WANG Yongfeng1,2, TIAN Hongcheng1, HU Rong1,2, WANG Yanrong1,3, PEI Xiuying1, XU Xian1,2   

  1. 1. Key Laboratory of Fertility Preservation and Maintenance, Ministry of Education;
    2. Fertility Center for Reproductive Medicine;
    3. Department of Anatomy and Histology-embryology, Ningxia Medical University, Yinchuan 750004, Ningxia, China
  • Received:2015-04-30 Online:2016-05-16 Published:2016-05-16

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摘要： 目的 探讨雌激素(E2)对雌激素受体-β(ER-β)与转录因子叉头蛋白3(FoxO3)的表达及其对卵巢颗粒细胞增殖与凋亡的影响。 方法 利用添加1 μmol/L的E2或100 nmol/L ICI182.780的TCM199培养基体外培养卵巢颗粒细胞;采用 WST比色法检测不同培养条件下卵巢颗粒细胞的增殖; RT-PCR 和Real-time PCR检测ER-β与FoxO3 mRNA的表达,采用细胞免疫荧光和Western blotting检测ER-β与FoxO3的表达。 结果 E2能促进卵巢颗粒细胞增殖,而ICI182.780抑制卵巢颗粒细胞增殖;RT-PCR扩增结果与理论值符合,Real-time PCR结果显示,ICI182.780与E2比较,ER-β mRNA表达下调, FoxO3 mRNA表达上调(P<0.05);Western blotting法结果显示,ER-β和FoxO3蛋白表达差异有统计学意义(P<0.05),灰度值分析显示添加E2中ER-β蛋白表达升高,FoxO3蛋白下降(P<0.05),添加ICI182.780中的ER-β表达下调, FoxO3表达上调,与E2比较差异有统计学意义(P<0.05)。 结论 E2可提高ER-β的表达,降低FoxO3的表达,影响卵巢颗粒细胞的增殖与凋亡。

关键词: 雌激素受体-β, 转录因子叉头蛋白3, 卵巢颗粒细胞, 雌激素

Abstract: Objective To explore the effects of estradiol(E2)on estrogen receptor-β(ER-β)and forkhead transcription factor 3(FoxO3)expression in the proliferation and apoptosis of human granulosa cells during ovarian stimulation. Methods Ovarian granulosa cells in healthy ovarian follicles were cultured in TCM199 medium supplemented with 1 μmol /L E2, and 100 nmol/L ICI182.780. The proliferation of ovarian granulosa cells was detected with WST. The ER-β and FoxO3 mRNA expressions in granulosa cells were determined with RT-PCR and real-time PCR. The ER-β and FoxO3 protein expressions were assessed with immunohistochemistry and Western blotting. Results E2 significantly improved while ICI182.780 inhibited the proliferation of ovarian granulosa cells. Real-time PCR results indicated that the ER-β mRNA expression was downregulated, while FoxO3 mRNA expression was upregulated(P<0.05). Western 山 东 大 学 学 报 (医 学 版)54卷5期 -马会明,等.雌激素对卵巢颗粒细胞雌激素受体-β和转录因子叉头蛋白3表达的影响 \=-blotting results showed that changes of ER-β and FoxO3 protein expressions were statistically significant(P<0.05). Grey level analysis showed that ER-β protein expression increased and FoxO3 protein expression was reduced with addition of E2(P<0.05), while ER-β protein expression decreased and FoxO3 protein expression increased with addition of ICI182.780(P<0.05). Conclusion E2 can affect the proliferation and apoptosis of ovarian granulosa cells by increasing ER-β expression while decreasing FoxO3 expression.

Key words: Estradiol, Estrogen receptor-β, Forkhead transcription factor 3, Ovarian granulosa cells

中图分类号: 

  • R711.6
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