Human chorionic gonadotropin-induced amphiregulin stimulates aromatase expression in human granulosa-lutein cells:a mechanism for estradiol production in the luteal phase
Human chorionic gonadotropin-induced amphiregulin stimulates aromatase expression in human granulosa-lutein cells:a mechanism for estradiol production in the luteal phase
复制标题
人绒毛膜促性腺激素诱导的双调蛋白刺激人颗粒叶黄素细胞中芳香酶的表达：黄体期雌二醇产生的机制
DOI：
10.1093/humrep/dez171
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发表时间：
2019-10-01
影响因子：
6.1
通讯作者：
Sun, Ying-Pu
中科院分区：
文献类型：
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作者：
Fang, Lanlan;Yu, Yiping;Sun, Ying-Pu
研究问题：双调蛋白（AREG），卵泡液中最丰富和最重要的表皮生长因子受体（EGFR）配体，是否调节人颗粒黄体（hGL）细胞芳香化酶的表达？简要答复：AREG介导hCG诱导的hGL细胞中芳香化酶表达和雌二醇（E2）产生的上调。已知：hCG在hGL细胞中快速诱导AREG的表达和分泌，并介导卵巢中LH/hCG的生理功能。EGFR蛋白不仅在排卵前期而且在月经周期的整个黄体期在卵泡中表达。在LH激增后，人黄体分泌高水平的E2，其调节各种黄体细胞功能。芳香化酶是负责E2生物合成中关键步骤的酶。然而，AREG是否调节芳香化酶的表达和E2的生产在hGL cells仍然unexplored.Study设计，规模，持续时间：这项研究是一项实验研究，进行了1年的时间。在体外调查研究的芳香化酶的表达和E2的生产在原代hGL cells.PARTICIPANTS/MATERIALS，设置，方法：原代hGL细胞的调节AREG的作用，从接受IVF治疗的妇女在一个学术研究中心。将hGL细胞暴露于重组人AREG、hCG或LH后，检测芳香化酶mRNA和蛋白水平。采用EGFR酪氨酸激酶抑制剂AG 1478、P13 K抑制剂LY 294002和靶向EGFR、LH受体、SCAR和AREG的siRNA验证其作用的特异性并探讨其分子机制。RT-qPCR和Western blot分别检测特异性mRNA和蛋白水平。收集65例不孕妇女在IVF治疗过程中的卵泡液和血清。Pearson相关性分析进行检查两个values.Main结果和机会的作用之间的相关系数：AREG刺激的芳香化酶表达和E2生产的hGL细胞的治疗。使用药理学抑制剂和特异性siRNA，我们发现AREG刺激芳香酶表达和E2的产生通过EGFR介导的蛋白激酶B（AKT）信号通路的激活。此外，EGFR活性的抑制和AREG敲低减弱了hCG诱导的芳香化酶表达和E2产生的上调。重要的是，卵泡液中AREG的蛋白水平与取卵2天后血清中的E2水平和IVF患者卵泡液中的E2水平呈正相关。大规模数据：N/A。局限性，回避原因：本研究的体外设置是一个局限性，可能无法反映真实的卵巢内微环境。临床数据是从一个小的样本size.WIDER影响的调查结果：我们的研究结果提供了第一个证据表明，hCG诱导的AREG有助于芳香化酶的表达和E2的生产在月经周期的黄体期。更好地了解女性生殖功能的激素调节可能有助于开发临床不孕症治疗的新策略。
STUDY QUESTION: Does amphiregulin (AREG), the most abundant and important epidermal growth factor receptor (EGFR) ligand in the follicular fluid, regulate aromatase expression in human granulosa-lutein (hGL) cells?SUMMARY ANSWER: AREG mediates the hCG-induced up-regulation of aromatase expression and estradiol (E2) production in hGL cells.WHAT IS KNOWN ALREADY: AREG expression and secretion are rapidly induced by hCG in hGL cells and mediate physiological functions of LH/hCG in the ovary. EGFR protein is expressed in follicles not only in the pre-ovulatory phase but also throughout the luteal phase of the menstrual cycle. After the LH surge, the human corpus luteum secretes high levels of E2, which regulates various luteal cell functions. Aromatase is an enzyme responsible for a key step in the biosynthesis of E2. However, whether AREG regulates aromatase expression and E2 production in hGL cells remains unexplored.STUDY DESIGN, SIZE, DURATION: This study is an experimental study performed over a 1-year period. In vitro investigations examined the role of AREG in the regulation of aromatase expression and E2 production in primary hGL cells.PARTICIPANTS/MATERIALS, SETTING, METHODS: Primary hGL cells were obtained from women undergoing IVF treatment in an academic research center. Aromatase mRNA and protein levels were examined after exposure of hGL cells to recombinant human AREG, hCG or LH. The EGFR tyrosine kinase inhibitor AG 1478, P13K inhibitor LY294002 and siRNAs targeting EGFR, LH receptor, SCAR and AREG were used to verify the specificity of the effects and to investigate the underlying molecular mechanisms. Reverse transcription quantitative real-time PCR (RT-qPCR) and western blot were used to measure the specific mRNA and protein levels, respectively. Follicular fluid and serum were collected from 65 infertile women during IVF treatment. Pearson's correlation analysis was performed to examine the correlation coefficient between two values.MAIN RESULTS AND THE ROLE OF CHANCE: Treatment of hGL cells with AREG-stimulated aromatase expression and E2 production. Using pharmacological inhibitors and specific siRNAs, we revealed that AREG-stimulated aromatase expression and E2 production via EGFR-mediated activation of the protein kinase B (AKT) signaling pathway. In addition, inhibition of EGFR activity and AREG knockdown attenuated hCG-induced up-regulation of aromatase expression and E2 production. Importantly, the protein levels of AREG in the follicular fluid were positively correlated with the E2 levels in serum after 2 days of oocyte pick-up and in the follicular fluid of IVF patients.LARGE-SCALE DATA: N/A.LIMITATIONS, REASONS FOR CAUTION: The in vitro setting of this study is a limitation that may not reflect the real intra-ovarian microenvironment. Clinical data were obtained from a small sample size.WIDER IMPLICATIONS OF THE FINDINGS: Our results provide the first evidence that hCG-induced AREG contributes to aromatase expression and E2 production in the luteal phase of the menstrual cycle. A better understanding of the hormonal regulation of female reproductive function may help to develop new strategies for the treatment of clinical infertility.