College of Animal Science and Technology, Key Laboratory of Animal Genetic, Breeding and Reproduction in Shaanxi Province, Northwestern A&F University, Yangling, Shaanxi, China.
Reprod Domest Anim. 2021 Feb;56(2):324-332. doi: 10.1111/rda.13869. Epub 2020 Dec 10.
Follicle-stimulating hormone (FSH) plays a critical role in follicular growth and granulosa cell function; however, the mechanism by which the aggressive stimulation of FSH leads to poorer oocyte quality and embryo development potential is unclear. In this study, bovine ovarian granulosa cells (BGCs) were challenged with FSH doses (vehicle, 0.1, 1, 10 and 100 ng/ml) to investigate the effects of FSH on BGCs. The results indicated that the relative viability of BGCs was significantly increased in cells challenged with 1 ng/ml FSH, whereas the viability was significantly decreased with 100 ng/ml FSH treatment. The mRNA abundance of FSHR, CYP19, StAR and BAX was significantly upregulated with 1, 10 and 100 ng/ml of FSH, while the BCL-2 mRNA level was downregulated with higher concentrations of FSH (10 and 100 ng/ml). Furthermore, BGC autophagy was detected in cells treated with 10 and 100 ng/ml FSH by MDC staining, and the mRNA abundance of LC3, BECN1, BNIP3, ATG3 and ATG7 was upregulated with increasing FSH concentration. Meanwhile, the protein expression of LC3 was increased in cells treated with 10 and 100 ng/ml FSH. 1 and 10 ng/ml FSH significantly increased E2 production, whereas 10 and 100 ng/ml FSH significantly increased P4 production. FSH significantly inhibited the phosphorylation of AKT in cells treated with higher concentrations (1, 10 and 100 ng/ml), while activating mTOR phosphorylation at concentrations of 10 and 100 ng/ml of FSH. In summary, we can conclude that higher doses of FSH (10 and 100 ng/ml) induce BGC autophagy via the AKT/mTOR signalling pathway.
卵泡刺激素（FSH）在卵泡生长和颗粒细胞功能中起着关键作用；然而，FSH 的剧烈刺激导致卵母细胞质量和胚胎发育潜能变差的机制尚不清楚。在这项研究中，用 FSH 剂量（对照、0.1、1、10 和 100 ng/ml）刺激牛卵巢颗粒细胞（BGC），以研究 FSH 对 BGC 的影响。结果表明，1 ng/ml FSH 刺激的 BGC 相对活力显著增加，而 100 ng/ml FSH 处理则显著降低。1、10 和 100 ng/ml 的 FSH 使 FSHR、CYP19、StAR 和 BAX 的 mRNA 丰度显著上调，而高浓度的 FSH（10 和 100 ng/ml）使 BCL-2 mRNA 水平下调。此外，用 MDC 染色检测到 10 和 100 ng/ml FSH 处理的 BGC 自噬，随着 FSH 浓度的增加，LC3、BECN1、BNIP3、ATG3 和 ATG7 的 mRNA 丰度上调。同时，用 10 和 100 ng/ml FSH 处理的细胞中 LC3 蛋白表达增加。1 和 10 ng/ml FSH 显著增加 E2 的产生，而 10 和 100 ng/ml FSH 显著增加 P4 的产生。FSH 显著抑制高浓度（1、10 和 100 ng/ml）处理的细胞中 AKT 的磷酸化，而在 10 和 100 ng/ml 的 FSH 浓度下激活 mTOR 磷酸化。总之，我们可以得出结论，较高剂量的 FSH（10 和 100 ng/ml）通过 AKT/mTOR 信号通路诱导 BGC 自噬。
