Duan Hongwei, Wang Fang, Wang Ke, Yang Shuai, Zhang Rong, Xue Chen, Zhang Lihong, Ma Xiaofei, Du Xianghong, Kang Jian, Zhang Yong, Zhao Xingxu, Hu Junjie, Xiao Longfei
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, 730070, Gansu, China.
Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation, Lanzhou, 730070, Gansu, China.
J Anim Sci Biotechnol. 2024 Sep 5;15(1):119. doi: 10.1186/s40104-024-01078-5.
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 HO-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 HO 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.
卵泡囊肿是高产奶牛繁殖损失的重要原因。这是由氧化应激导致的卵泡颗粒细胞（GCs）死亡引起的。已知槲皮素具有显著的抗氧化和抗凋亡作用。然而，槲皮素对卵泡囊肿的影响尚未阐明。因此，本研究旨在探讨槲皮素在过氧化氢（HO）诱导的原代奶牛GCs和3-硝基丙酸（3-NPA）诱导的小鼠氧化应激模型中的抗氧化和抗凋亡作用及潜在分子机制，从而治疗奶牛卵巢囊肿。
在本研究中，与发情奶牛相比，患有卵泡囊肿的奶牛氧化应激水平升高，卵泡细胞凋亡增加，而自噬水平降低。HO在体外诱导氧化应激模型，导致凋亡显著增加，同时自噬减少。槲皮素显著改善了这些作用。在用活性氧（ROS）抑制剂N-乙酰半胱氨酸（NAC）处理细胞后，观察到了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）、西利司他（SIRT1抑制剂）和化合物C（AMPK抑制剂）的进一步研究表明，氯喹抵消了槲皮素对氧化应激诱导凋亡的作用，而雷帕霉素与槲皮素具有相同的效果。此外，SIRT1/AMPK途径抑制剂拮抗了槲皮素介导的氧化应激对凋亡增加和自噬减少影响的减轻作用。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，同时减轻小鼠卵巢的氧化应激损伤并抑制凋亡。
这些发现表明，槲皮素可通过SIRT1/ROS/AMPK信号通路激活自噬来抑制GCs凋亡并恢复卵巢功能，为高产奶牛卵巢卵泡囊肿的治疗提供了新方向。
