Regulation of AMH by oocyte-specific growth factors in human primary cumulus cells.
Regulation of AMH by oocyte-specific growth factors in human primary cumulus cells.
复制标题
DOI：
10.1530/rep-17-0421
复制
发表时间：
2017-12
期刊：
影响因子：
--
通讯作者：
Stocco C
中科院分区：
文献类型：
--
作者：
Convissar S;Armouti M;Fierro MA;Winston NJ;Scoccia H;Zamah AM;Stocco C
卵泡细胞对AMH产生的调节知之甚少。本研究的目的是确定卵母细胞分泌的因子，生长分化因子9（GDF 9）和骨形态发生蛋白15（BMP 15），对AMH的生产在原代人卵丘细胞的作用。将IVF患者的卵丘细胞与GDF 9、BMP 15、重组FSH和特异性信号传导抑制剂的组合培养。GDF 9或BMP 15单独刺激对AMH mRNA水平无显著影响。相反，同时刺激GDF 9和BMP 15（G+B）导致AMH mRNA表达显著增加。增加G+B浓度（0.6、2.5、5和10 ng/ml）以剂量依赖方式刺激AMH，在5 ng/ml时显示最大效应。Western印迹分析显示，与对照组相比，G+B处理的细胞中AMH蛋白水平平均增加16倍。FSH联合处理降低G+B对AMH表达的刺激。G+B对AMH表达的刺激作用可被SMAD 2/3信号通路的抑制剂显著降低。这些研究结果首次表明，AMH的生产是由卵母细胞分泌的因子在原代人类卵丘细胞调节。此外，我们的新发现证实GDF 9 + BMP 15的组合有效地刺激AMH表达。
The regulation of AMH production by follicular cells is poorly understood. The purpose of this study was to determine the role of the oocyte-secreted factors, growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), on AMH production in primary human cumulus cells. Cumulus cells from IVF patients were cultured with a combination of GDF9, BMP15, recombinant FSH, and specific signaling inhibitors. Stimulation with GDF9 or BMP15 separately had no significant effect on AMH mRNA levels. In contrast, simultaneous stimulation with GDF9 and BMP15 (G+B) resulted in a significant increase in AMH mRNA expression. Increasing concentration of G+B (0.6, 2.5, 5 and 10 ng/ml) stimulated AMH in a dose-dependent manner, showing a maximal effect at 5 ng/ml. Western blot analyses revealed an average 16-fold increase in AMH protein levels in cells treated with G+B when compared to controls. FSH co-treatment decreased the stimulation of AMH expression by G+B. The stimulatory effect of G+B on the expression of AMH was significantly decreased by inhibitors of the SMAD2/3 signaling pathway. These findings show for the first time that AMH production is regulated by oocyte-secreted factors in primary human cumulus cells. Moreover, our novel findings establish that the combination of GDF9 + BMP15 potently stimulates AMH expression.