a Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Education Ministry of China, College of Animal Science and Technology , Huazhong Agricultural University , Wuhan 430070 , China.
b School of Veterinary Medicine , Wollega University , PO Box: 395, Nekemte , Ethiopia.
Cell Cycle. 2018;17(7):892-902. doi: 10.1080/15384101.2018.1456297. Epub 2018 Apr 24.
Recent findings suggest that ephrinA5 (Efna5) has a novel role in female mouse fertility, in addition to its well-defined role as a neurogenesis factor. Nevertheless, its physiological roles in ovarian granulosa cells (GC) have not been determined. In this study, mouse GC were cultured and transfected with ephrin A5 siRNA and negative control to determine the effects of Efna5 on GC apoptosis, proliferation, cell cycle progression, and related signaling pathways. To understand the mode signaling, the mRNA expression levels of Efna5 receptors (Eph receptor A5, Eph receptor A3, Eph receptor A8, and Eph receptor B2) were examined. Both mRNA and protein expressions of apoptosis-related factors (Bax, Bcl-2, Caspase 8, Caspase 3, and Tnfα) and a proliferation marker, Pcna, were investigated. Additionally, the role of Efna5 on paracrine oocyte-secreted factors and steroidogenesis hormones were also explored. Efna5 silencing suppressed GC apoptosis by downregulating Bax and upregulating Bcl-2 in a Caspase 8-dependent manner. Efna5 knockdown promoted GC proliferation via p-Akt and p-ERK pathway activation. The inhibition of Efna5 enhanced BMH15 and estradiol expression, but suppressed GDF9, while progesterone level remained unaltered. These results demonstrated that Efna5 is a pro-apoptotic agent in GC and plays important role in folliculogenesis by mediating apoptosis, proliferation, and steroidogenesis in female mouse. Therefore Efna5 might be potential therapeutic target for female fertility disorders.
最近的研究结果表明，除了作为神经发生因子的明确作用外，ephrinA5（Efna5）在雌性小鼠生育力中具有新的作用。然而，其在卵巢颗粒细胞（GC）中的生理作用尚未确定。在这项研究中，培养了小鼠 GC，并转染了 Ephrin A5 siRNA 和阴性对照，以确定 Efna5 对 GC 凋亡、增殖、细胞周期进程和相关信号通路的影响。为了了解信号转导模式，检查了 Ephrin A5 受体（Eph 受体 A5、Eph 受体 A3、Eph 受体 A8 和 Eph 受体 B2）的 mRNA 表达水平。研究了凋亡相关因子（Bax、Bcl-2、Caspase 8、Caspase 3 和 Tnfα）和增殖标志物 Pcna 的 mRNA 和蛋白表达。此外，还探讨了 Efna5 对旁分泌卵母细胞分泌因子和类固醇生成激素的作用。Efna5 沉默通过下调 Caspase 8 依赖性的 Bax 并上调 Bcl-2 来抑制 GC 凋亡。Efna5 敲低通过激活 p-Akt 和 p-ERK 通路促进 GC 增殖。Efna5 抑制增强了 BMH15 和雌二醇的表达，但抑制了 GDF9，而孕激素水平保持不变。这些结果表明，Efna5 是 GC 中的促凋亡剂，通过调节雌性小鼠的凋亡、增殖和类固醇生成在卵泡发生中发挥重要作用。因此，Efna5 可能是女性生育障碍的潜在治疗靶点。
