Regulation of Insulin-Like Growth Factor 2 by Oocyte-Secreted Factors in Primary Human Granulosa Cells
Regulation of Insulin-Like Growth Factor 2 by Oocyte-Secreted Factors in Primary Human Granulosa Cells
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DOI：
10.1210/clinem/dgz057
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发表时间：
2020-01-01
影响因子：
5.8
通讯作者：
Stocco, Carlos
中科院分区：
文献类型：
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作者：
Hobeika, Elie;Armouti, Marah;Stocco, Carlos
背景：人颗粒细胞(HGC)产生并应答胰岛素样生长因子2(IGF2)，但卵母细胞是否参与IGF2的调控尚不清楚。目的：探讨卵母细胞分泌因子(OSFs)如生长分化因子9(GDF9)和骨形态发生蛋白15(BMP15)在hGC产生IGF2中的作用。设计：原代培养的人卵丘GCs。地点：大学不孕不育中心。患者或其他参与者：体外受精的GCS。干预(S)：卵泡刺激素(FSH)、二丁基环磷酸腺苷(DBcAMP)、卵泡刺激素(FSH)和骨形态发生蛋白15(DBAMP)处理的细胞。结果：联合应用GDF9和BMP15(G+B)可显著增强FSH对IGF2的刺激作用(P<；P<；0.0001)，且呈浓度依赖关系，在浓度为5 ng/m L时效果最好。然而，GDF9或BMP15单独或联合(G+B)在没有FSH的情况下对IGF2没有影响。FSH刺激IGF2启动子3活性，而G+B对启动子活性无影响。G+B可增强cAMP对IGF2的刺激作用。Smad3抑制剂可抑制FSH对G+B刺激IGF2的增强作用(P<；0.05)，但不影响FSH的诱导作用。此外，抑制胰岛素样生长因子受体部分阻断了G+B对促性腺激素作用的增强作用(P<；0.009)。结论：我们首次发现卵母细胞主动参与调节hGC中IGF2的表达，这一作用是通过Smad2/3与G+B的特异性结合而介导的，进而靶向FSH受体下游的机制。
Context: Human granulosa cells (hGCs) produce and respond to insulin-like growth factor 2 (IGF2) but whether the oocyte participates in IGF2 regulation in humans is unknown.Objective: To determine the role of oocyte-secreted factors (OSFs) such as growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) in IGF2 production by hGCs.Design: Primary human cumulus GCs in culture.Setting: University infertility center.Patients or Other Participants: GCs of women undergoing in vitro fertilization.Intervention(s): Cells treated with GDF9 and BMP15 in the presence of vehicle, follicle-stimulating hormone (FSH), dibutyryl cyclic-AMP (dbcAMP), or mothers against decapentaplegic homolog (SMAD) inhibitors.Main Outcome Measure(s): Quantification of mRNA, protein, promoter activity, and DNA methylation.Results: FSH stimulation of IGF2 (protein and mRNA) was significantly potentiated by the GDF9 and BMP15 (G+B) combination (P < 0.0001) in a concentration-dependent manner showing a maximal effect at 5 ng/mL each. However, GDF9 or BMP15 alone or in combination (G+B) have no effect on IGF2 in the absence of FSH. FSH stimulated IGF2 promoter 3 activity, but G+B had no effect on promoter activity. G+B potentiated IGF2 stimulation by cAMP. SMAD3 inhibitors inhibited G+B enhancement of IGF2 stimulation by FSH (P < 0.05) but had no effect on FSH induction. Moreover, inhibition of insulin-like growth factor receptor partially blocked G+B potentiation of FSH actions (P < 0.009).Conclusions: For the first time, we show that the oocyte actively participates in the regulation of IGF2 expression in hGCs, an effect that is mediated by the specific combination of G+B via SMAD2/3, which in turn target mechanisms downstream of the FSH receptor.