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Abstract：
Within the poultry industry, hens\' reproductive performance is of great economic significance. The development and growth of follicles is a key aspect of hen egg production, and ovarian follicle growth and development are closely associated with granulosa cells (GCs) proliferation and the synthesis of steroid hormones. It has been confirmed by numerous studies that microRNAs (miRNAs) play important roles in the steroid hormone synthesis and proliferation of GCs. In this study, we examined the main miRNAs influencing hens\' ability to reproduce, identified the miR-223 that is mainly expressed in atretic follicles based on sequencing, and investigated its role in GCs. Then, we used miR-223 mimic and inhibitor to knockdown or overexpress miR-223 expression. The result showed that miR-223 significantly inhibits both the steroid hormone synthesis and the proliferation of GCs. Subsequently, the results of the dual luciferase reporter experiment and bioinformatics prediction demonstrated that cysteine rich transmembrane BMP regulator 1 (CRIM1) was a downstream target gene of miR-223, and overexpression of miR-223 prevented CRIM1 expression. The function of CRIM1 was further investigated, and we observed a significant reduction in the synthesis of steroid hormones and the proliferation of GCs after transfection with CRIM1 siRNA. The opposite function of miR-223 was observed for CRIM1 in our study. Additionally, we demonstrated the involvement of the miR-223/CRIM1 axis in GCs through modulation of the AKT signaling pathway. Our data demonstrate the pivotal role of the miR-223 in the proliferation and steroid hormone synthesis of chicken GCs, which helps to explain how non-coding RNA (ncRNA) affects chicken reproductive function.
摘要：
在家禽业中，母鸡的繁殖性能具有重要的经济意义。卵泡的发育和生长是鸡蛋生产的关键方面，卵泡的生长发育与颗粒细胞(GCs)的增殖和类固醇激素的合成密切相关。大量研究证实,microRNAs(miRNAs)在类固醇激素合成和GCs增殖中起重要作用。在这项研究中,我们检查了影响母鸡繁殖能力的主要miRNA，基于测序鉴定了主要在闭锁卵泡中表达的miR-223，并研究了其在GC中的作用。然后，我们使用miR-223模拟物和抑制剂敲低或过表达miR-223.结果显示miR-223显著抑制类固醇激素的合成和GCs的增殖。随后，双荧光素酶报告基因实验和生物信息学预测结果表明,富含半胱氨酸的跨膜BMP调节因子1(CRIM1)是miR-223的下游靶基因,miR-223的过表达阻止了CRIM1的表达.进一步研究了CRIM1的功能，我们观察到用CRIM1siRNA转染后，类固醇激素的合成和GC的增殖显着减少。在我们的研究中,对于CRIM1观察到miR-223的相反功能。此外,我们证明miR-223/CRIM1轴通过调节AKT信号通路参与GCs.我们的数据证明了miR-223在鸡GC的增殖和类固醇激素合成中的关键作用，这有助于解释非编码RNA（ncRNA）如何影响鸡的繁殖功能。
