Ma Jianyu, Yang Hua, Liu Liang, Wan Yongjie, Wang Feng
Institute of Sheep and Goat Science; Nanjing Agricultural University, Nanjing, 210095, China.
Institute of Sheep and Goat Science; Nanjing Agricultural University, Nanjing, 210095, China.
Theriogenology. 2021 Oct 1;173:83-92. doi: 10.1016/j.theriogenology.2021.07.011. Epub 2021 Jul 24.
Energy balance is essential for normal reproduction of ram. However, the effect of energy restriction (ER) on reactive oxygen species (ROS) of sheep Leydig cells (LCs) and the rescuee methods are still unclear. To investigate the in vitro effect of melatonin on cellular ROS in fER-treated sheep LCs and explore the underlying mechanism, Hu sheep LCs were restricted energy using no serum culture medium and resaved with 10 ng/ml melatonin, respectively. The results showed that ER significantly increased MDA level, while decreased CAT, GHS-px expression and ΔΨm (p < 0.05). Meanwhile, ER decreased testosterone concentration and cell proliferation rate (p < 0.05). And the expression of testosterone synthesis-related enzymes was also down-regulated by ER (p < 0.05). Furthermore, we revealed that melatonin reversed the defective phenotypes in ER-treated LCs via Sirt1/Sod2 pathway. The interference of Sirt1 abolished the melatonin-mediated improvement of cellular ROS and testosterone secretion. Taken together, our study firstly indicated that melatonin could alleviate the excessive ROS accumulation and promote testosterone biosynthesis in ER-treated sheep LCs via the activation of Sirt1/Sod2 pathway.
能量平衡对于公羊的正常繁殖至关重要。然而，能量限制（ER）对绵羊睾丸间质细胞（LCs）活性氧（ROS）的影响以及挽救方法仍不清楚。为了研究褪黑素对经fER处理的绵羊LCs细胞内ROS的体外作用并探索其潜在机制，分别使用无血清培养基对湖羊LCs进行能量限制并用10 ng/ml褪黑素进行挽救。结果表明，ER显著提高了丙二醛水平，同时降低了CAT、GHS-px表达和线粒体膜电位（p < 0.05）。同时，ER降低了睾酮浓度和细胞增殖率（p < 0.05）。ER还下调了睾酮合成相关酶的表达（p < 0.05）。此外，我们发现褪黑素通过Sirt1/Sod2途径逆转了经ER处理的LCs中的缺陷表型。Sirt1的干扰消除了褪黑素介导的细胞ROS改善和睾酮分泌。综上所述，我们的研究首次表明，褪黑素可以通过激活Sirt1/Sod2途径减轻经ER处理的绵羊LCs中过量的ROS积累并促进睾酮生物合成。
