miR-375 mediates CRH signaling pathway in inhibiting E2 synthesis in porcine ovary
miR-375 mediates CRH signaling pathway in inhibiting E2 synthesis in porcine ovary
复制标题
MiR-375 介导 CRH 信号通路抑制猪卵巢 E2 合成。
DOI：
10.1530/rep-16-0323
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发表时间：
2017-01-01
期刊：
影响因子：
3.8
通讯作者：
Cui, Sheng
中科院分区：
文献类型：
--
作者：
Yu, Chulin;Li, Meiling;Cui, Sheng
促肾上腺皮质激素释放激素（CRH）信号系统参与许多应激相关的生理和病理反应，包括抑制卵巢雌二醇（E2）的合成和卵泡发育。此外，有报道称microRNAs（miRNAs）可以控制动物生殖系统的功能。本研究旨在探讨miR-375在猪卵巢中的功能及其与CRH信号分子的关系。首先，我们的普通PCR结果显示，miR-375和CRH受体1（CRHR 1）在猪卵巢中表达，而CRH受体2（CRHR 2）未检测到。我们进一步通过原位杂交（ISH）对miR-375和CRHR 1的细胞类型进行了定位，结果显示miR-375仅定位于颗粒细胞，而CRHR 1在所有颗粒细胞和卵母细胞中均呈阳性，推测miR-375和CRHR 1共定位于颗粒细胞。其次，我们发现在培养的颗粒细胞中过表达miR-375抑制E2的产生，而miR-375敲低则显示相反的结果。此外，我们的体外研究结果表明，miR-375介导的CRH抑制E2合成的信号通路。最后，我们的数据表明，miR-375的作用是通过直接结合特异性蛋白1（SP1）mRNA的3 'UTR来降低SP1蛋白水平。因此，我们得出结论，miR-375是通过介导CRH信号通路调节E2合成的关键因子。
The corticotropin-releasing hormone (CRH) signaling system is involved in numbers of stress-related physiological and pathological responses, including its inhibiting effects on estradiol (E2) synthesis and follicular development in the ovary. In addition, there are reports that microRNAs (miRNAs) can control the function of animal reproductive system. The aim of present study was to investigate the functions of miR-375 and the relationship between miR-375 and CRH signaling molecules in the porcine ovary. First, our common PCR results show that miR-375 and the CRH receptor 1 (CRHR1) are expressed in porcine ovary, whereas CRH receptor 2 (CRHR2) is not detected. We further have located the cell types of miR-375 and CRHR1 by in situ hybridization (ISH), and the results show that miR-375 is located only in the granulosa cells, whereas CRHR1 is positive in all of granulosa cells and oocytes, inferring that miR-375 and CRHR1 are co-localized in granulosa cells. Second, we show that overexpression of miR-375 in cultured granulosa cells suppresses the E2 production, whereas miR-375 knockdown demonstrates the opposite result. Besides, our in vitro results demonstrate that miR-375 mediates the signaling pathway of CRH inhibiting E2 synthesis. Finally, our data show that the action of miR-375 is accomplished by directly binding to the 3'UTR of specificity protein1 (SP1) mRNA to decrease the SP1 protein level. Thus, we conclude that miR-375 is a key factor in regulating E2 synthesis by mediating the CRH signaling pathway.