Suppression of Notch Signaling Stimulates Progesterone Synthesis by Enhancing the Expression of NR5A2 and NR2F2 in Porcine Granulosa Cells
Suppression of Notch Signaling Stimulates Progesterone Synthesis by Enhancing the Expression of NR5A2 and NR2F2 in Porcine Granulosa Cells
复制标题
抑制 Notch 信号传导通过增强猪颗粒细胞中 NR5A2 和 NR2F2 的表达来刺激孕酮合成
DOI：
10.3390/genes11020120
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发表时间：
2020-02-01
期刊：
影响因子：
3.5
通讯作者：
Shi, Zhendan
中科院分区：
文献类型：
--
作者：
Guo, Rihong;Chen, Fang;Shi, Zhendan
据报道，保守的Notch途径参与了孕酮的合成和分泌；然而，确切的影响仍然存在争议。为了探讨Notch信号通路在猪颗粒细胞孕酮生物合成中的作用和可能的机制，我们首先用药理上的伽玛分泌酶抑制剂N-(N-(3，5-difluorophenacetyl-l-alanyl))-S-phenylglycine叔丁酯阻断了培养的PGCs中的Notch信号通路，然后研究了孕酮生物合成途径和调节类固醇合成的关键转录因子的基因表达。我们发现DAPT剂量和时间依赖地增加孕酮的分泌。类固醇合成蛋白NPC1和STAR以及两个转录因子NR5A2和NR2F2的表达显著上调，而HSD3B的表达显著下调。此外，用特异性siRNAs敲除NR5A2和NR2F2均可阻断DAPT对孕激素分泌的上调作用，并逆转DAPT对NPC1、STAR和HSD3B表达的影响。此外，NR5A2和NR2F2的敲除可刺激Notch3的表达。结论：抑制Notch信号通过增强NPC1和STAR的表达来促进孕酮的分泌，NR5A2和NR2F2是Notch信号的下游因子，参与调节孕酮的合成。
The conserved Notch pathway is reported to be involved in progesterone synthesis and secretion; however, the exact effects remain controversial. To determine the role and potential mechanisms of the Notch signaling pathway in progesterone biosynthesis in porcine granulosa cells (pGCs), we first used a pharmacological gamma-secretase inhibitor, N-(N-(3,5-difluorophenacetyl-l-alanyl))-S-phenylglycine t-butyl ester (DAPT), to block the Notch pathway in cultured pGCs and then evaluated the expression of genes in the progesterone biosynthesis pathway and key transcription factors (TFs) regulating steroidogenesis. We found that DAPT dose- and time-dependently increased progesterone secretion. The expression of steroidogenic proteins NPC1 and StAR and two TFs, NR5A2 and NR2F2, was significantly upregulated, while the expression of HSD3B was significantly downregulated. Furthermore, knockdown of both NR5A2 and NR2F2 with specific siRNAs blocked the upregulatory effects of DAPT on progesterone secretion and reversed the effects of DAPT on the expression of NPC1, StAR, and HSD3B. Moreover, knockdown of NR5A2 and NR2F2 stimulated the expression of Notch3. In conclusion, the inhibition of Notch signaling stimulated progesterone secretion by enhancing the expression of NPC1 and StAR, and the two TFs NR5A2 and NR2F2 acted as downstream TFs of Notch signaling in regulating progesterone synthesis.