Follicular cysts contribute significantly to reproductive loss in high-yield dairy cows. This results from the death of follicular granulosa cells (GCs) caused by oxidative stress. Quercetin is known to have significant antioxidant and anti-apoptotic effects. However, the effect of quercetin on follicular cysts has yet been elucidated. Therefore, this study aimed to explore the anti-oxidant and anti-apoptosis effects and potential molecular mechanisms of quercetin in H2O2-induced primary cow GCs and 3-nitropropionic acid (3-NPA)-induced mouse model of oxidative stress and thus treat ovarian cysts in dairy cows. In this study, compared with estrus cows, cows with follicular cysts showed heightened levels of oxidative stress and increased follicular cell apoptosis, while autophagy levels were reduced. A model of oxidative stress was induced in vitro by H2O2 and showed significant increases in apoptosis together with reduced autophagy. These effects were significantly ameliorated by quercetin. Effects similar to those of quercetin were observed after treatment of cells with the reactive oxygen species (ROS) inhibitor N-acetylcysteine (NAC). Further investigations using chloroquine (autophagy inhibitor), rapamycin (autophagy activator), selisistat (SIRT1 inhibitor), and compound C (AMPK inhibitor) showed that chloroquine counteracted the effects of quercetin on oxidative stress-induced apoptosis, while rapamycin had the same effect as quercetin. In addition, the SIRT1/AMPK pathway inhibitors antagonized quercetin-mediated mitigation of the effects of oxidative stress on increased apoptosis and reduced autophagy. Consistent with the results in vitro, in mouse ovarian oxidative stress model induced by 3-NPA, quercetin activated autophagy through the SIRT1/AMPK signaling pathway, while alleviating oxidative stress damage and inhibiting apoptosis in mouse ovaries. These findings indicate that quercetin can inhibit apoptosis in GCs and restore ovarian function by activating autophagy through the SIRT1/ROS/AMPK signaling pathway, suggesting a new direction for the treatment of ovarian follicular cysts in high-yield dairy cows.

中文翻译：

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槲皮素通过 SIRT1/ROS/AMPK 信号通路激活自噬，改善氧化应激诱导的奶牛滤泡囊肿颗粒细胞凋亡


滤泡囊肿会严重导致高产奶牛的繁殖损失。这是由于氧化应激导致滤泡颗粒细胞（GC）死亡的结果。已知槲皮素具有显着的抗氧化和抗细胞凋亡作用。然而，槲皮素对卵泡囊肿的作用尚未阐明。因此，本研究旨在探讨槲皮素在H2O2诱导的原代牛GC和3-硝基丙酸（3-NPA）诱导的氧化应激小鼠模型中的抗氧化和抗凋亡作用及其潜在分子机制，从而治疗卵巢癌奶牛的囊肿。在这项研究中，与发情奶牛相比，患有滤泡囊肿的奶牛表现出氧化应激水平升高，滤泡细胞凋亡增加，而自噬水平降低。 H2O2 在体外诱导氧化应激模型，显示细胞凋亡显着增加，同时自噬减少。槲皮素可显着改善这些影响。用活性氧 (ROS) 抑制剂 N-乙酰半胱氨酸 (NAC) 处理细胞后，观察到与槲皮素类似的效果。使用氯喹（自噬抑制剂）、雷帕霉素（自噬激活剂）、selisistat（SIRT1抑制剂）和化合物C（AMPK抑制剂）进一步研究表明，氯喹抵消了槲皮素对氧化应激诱导的细胞凋亡的影响，而雷帕霉素具有相同的效果槲皮素。此外，SIRT1/AMPK 通路抑制剂可以拮抗槲皮素介导的氧化应激对细胞凋亡增加和自噬减少的影响。 与体外结果一致，在3-NPA诱导的小鼠卵巢氧化应激模型中，槲皮素通过SIRT1/AMPK信号通路激活自噬，同时减轻氧化应激损伤并抑制小鼠卵巢细胞凋亡。这些研究结果表明，槲皮素可以通过SIRT1/ROS/AMPK信号通路激活自噬，抑制GC细胞凋亡，恢复卵巢功能，为高产奶牛卵巢滤泡囊肿的治疗提供新方向。