College of Animal Science and Technology, Ningxia University, Yinchuan, PR China.
College of Animal Science and Technology, Ningxia University, Yinchuan, PR China.
Steroids. 2024 Feb;202:109349. doi: 10.1016/j.steroids.2023.109349. Epub 2023 Dec 9.
RFRP-3 is a functional ortholog of avian GnIH and regulates reproductive activities in the gonads of animals. However, the role of RFRP-3 in the function of ovarian granulosa cells in mice remains unclear. First, we detected the expression of the RFRP-3 receptor (GPR147) in the ovarian granulosa cells of mice. Second, the effect of RFRP-3 treatment on estradiol and progesterone secretions from granulosa cells was tested by ELISA. Meanwhile, the expression of genes and proteins regulating steroid hormone synthesis was respectively examined by qPCR and western blot. Furthermore, the effect of RFRP-3 treatment on the apoptosis of granulosa cells was analyzed. The results revealed that the GPR147 protein (a RFRP-3 receptor) was expressed in the ovarian granulosa cells of mice. Low and medium doses RFRP-3 treatment significantly reduced progesterone secretion in the granulosa cells (P < 0.05), while RFRP-3 suppressed p450scc, 3β-HSD, StAR, and FSHR expression in a non-dose-dependent manner. Moreover, RFRP-3 treatment might induce the apoptosis of granulosa cells. Additionally, low doses RFRP-3 significantly reduced p-ERK1/2 protein expression (P < 0.05) in the ovarian granulosa cells. We here, for the first time, confirmed that GPR147 was expressed in the ovarian granulosa cells of mice. Our findings suggested that and RFRP-3 regulates the granulosa cell function through the ERK signaling pathway, which will lay the foundation for uncovering molecular mechanisms by which RFRP-3 regulates follicle development in future.
RFRP-3 是一种功能性的禽类 GnIH 同源物，调节动物性腺中的生殖活动。然而，RFRP-3 在小鼠卵巢颗粒细胞中的功能作用尚不清楚。首先，我们检测了 RFRP-3 受体（GPR147）在小鼠卵巢颗粒细胞中的表达。其次，通过 ELISA 检测了 RFRP-3 处理对颗粒细胞中雌二醇和孕酮分泌的影响。同时，通过 qPCR 和 Western blot 分别检测了调节甾体激素合成的基因和蛋白质的表达。此外，还分析了 RFRP-3 处理对颗粒细胞凋亡的影响。结果表明，GPR147 蛋白（RFRP-3 受体）在小鼠卵巢颗粒细胞中表达。低剂量和中剂量 RFRP-3 处理显著降低了颗粒细胞中孕酮的分泌（P<0.05），而 RFRP-3 以非剂量依赖的方式抑制了 p450scc、3β-HSD、StAR 和 FSHR 的表达。此外，RFRP-3 处理可能诱导颗粒细胞凋亡。此外，低剂量 RFRP-3 显著降低了卵巢颗粒细胞中 p-ERK1/2 蛋白的表达（P<0.05）。我们首次证实 GPR147 在小鼠卵巢颗粒细胞中表达。我们的研究结果表明，RFRP-3 通过 ERK 信号通路调节颗粒细胞功能，这将为揭示 RFRP-3 调节卵泡发育的分子机制奠定基础。
