Xu Huitao, Khan Adnan, Zhao Shanjiang, Wang Huan, Zou Huiying, Pang Yunwei, Zhu Huabin
Embryo Biotechnology and Reproduction Laboratory, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Key Laboratory of Animal Genetics, Breeding, and Reproduction, MARA, National Engineering Laboratory for Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
Animals (Basel). 2020 Feb 24;10(2):367. doi: 10.3390/ani10020367.
Inhibin A is well known for its inhibitory properties against follicle-stimulating hormone (FSH), released through a pituitary-gonadal negative feedback loop to regulate follicular development. Ovarian folliculogenesis, hormonal biosynthesis, and gametogenesis are dependent on inhibins, playing vital roles in promoting or inhibiting cell proliferation. The present study explored the physiological and molecular response of bovine granulosa cells (GCs) to different concentrations of inhibin A in vitro. We treated the primary GCs isolated from ovarian follicles (3-6 mm) with different levels of inhibin A (20, 50, and 100 ng/mL) along with the control (0 ng/mL) for 24 h. To evaluate the impact of inhibin A on GCs, several in vitro cellular parameters, including cell apoptosis, viability, cell cycle, and mitochondrial membrane potential (MMP) were detected. Besides, the transcriptional regulation of pro-apoptotic (BAX, Caspase-3) and cell proliferation (PCNA, CyclinB1) genes were also quantified. The results indicated a significant ( < 0.05) increase in the cell viability in a dose-dependent manner of inhibin A. Likewise, MMP was significantly ( < 0.05) enhanced when GCs were treated with high doses (50, 100 ng/mL) of inhibin A. Furthermore, inhibin A dose (100 ng/mL) markedly improved the progression of the G1 phase of the cell cycle and increased the cell number in the S phase, which was supported by the up-regulation of the proliferating cell nuclear antigen PCNA (20, 50, and 100ng/mL) and CyclinB (100 ng/mL) genes. In addition, higher doses of inhibin A (50 and 100 ng/mL) significantly ( < 0.05) decreased the apoptotic rate in GCs, which was manifested by down regulating BAX and Caspase-3 genes. Conclusively, our study presented a worthy strategy for the first time to characterize the cellular adaptation of bovine GCs under different concentrations of inhibin A. Our results conclude that inhibin A is a broad regulatory marker in GCs by regulating apoptosis and cellular progression.
抑制素A以其对促卵泡激素(FSH)的抑制特性而闻名,它通过垂体-性腺负反馈回路释放,以调节卵泡发育。卵巢卵泡发生、激素生物合成和配子发生都依赖于抑制素,其在促进或抑制细胞增殖中发挥着至关重要的作用。本研究在体外探讨了牛颗粒细胞(GCs)对不同浓度抑制素A的生理和分子反应。我们用不同水平的抑制素A(20、50和100 ng/mL)以及对照(0 ng/mL)处理从卵巢卵泡(3-6毫米)分离的原代GCs 24小时。为了评估抑制素A对GCs的影响,检测了几个体外细胞参数,包括细胞凋亡、活力、细胞周期和线粒体膜电位(MMP)。此外,还对促凋亡基因(BAX、Caspase-3)和细胞增殖基因(PCNA、CyclinB1)的转录调控进行了定量分析。结果表明,抑制素A以剂量依赖性方式显著(<0.05)提高了细胞活力。同样,当GCs用高剂量(50、100 ng/mL)的抑制素A处理时,MMP显著(<0.05)增强。此外,抑制素A剂量(100 ng/mL)显著改善了细胞周期G1期的进程,并增加了S期的细胞数量,增殖细胞核抗原PCNA(20、50和100 ng/mL)和CyclinB(100 ng/mL)基因的上调支持了这一点。此外,高剂量的抑制素A(50和100 ng/mL)显著(<0.05)降低了GCs中的凋亡率,这通过下调BAX和Caspase-3基因得以体现。总之,我们的研究首次提出了一种有价值的策略,以表征不同浓度抑制素A作用下牛GCs的细胞适应性。我们的结果表明,抑制素A通过调节细胞凋亡和细胞进程,是GCs中的一种广泛的调节标志物。