Tang Ying, Wang Ziming, Chen Yanru, Wang Junying, Wang Hongzhan, Li Bo, Liu Bojing, Zheng Peng
Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China.
Genes (Basel). 2024 Nov 28;15(12):1544. doi: 10.3390/genes15121544.
: Oxidative stress in the testicles of male livestock can cause reduced fertility. Melatonin is a natural product with antioxidant effects, but its specific antioxidant mechanism is still unclear. This study used calf testicular Sertoli cells as materials to explore the mechanism by which melatonin alleviates the oxidative stress of Sertoli cells, laying a foundation for improving the fertility of bulls. : The optimal treatment concentrations of HO and melatonin (MLT) were screened out using a CCK8 kit and MDA kit. Then, the cells were divided into four groups for treatment: control group, HO treatment group, MLT treatment group, and HO and MLT co-treatment group, then the MDA, ROS, GSH, and SOD contents were detected. Real-time quantitative PCR analysis and Western blot analysis were used to detect genes and proteins related to the Nrf2-Keap1 pathway. Immunofluorescence staining was used to analyze changes in Nrf2. : Research results show that the MDA content of cells in the group treated with HO and MLT combined was significantly lower than that in the group treated with HO alone, but there was no difference from the control group. Compared with the control group, the ROS level of cells in the HO-treated group significantly increased, and the content of GSH and SOD significantly decreased. Compared with the HO-treated group, the ROS level of cells in the HO and MLT co-treated group significantly decreased, and the content of GSH and SOD increased significantly, but no difference from the control group. Similarly, MTL can alleviate the changes in cellular Nrf2, Keap1, HO-1, and NQO1 expression caused by HO. : Melatonin activates the Nrf2-Keap1 signaling pathway in Sertoli cells, elevating the expression of HO-1 and NQO1, and thereby exerting its antioxidant capabilities.
雄性家畜睾丸中的氧化应激会导致生育能力下降。褪黑素是一种具有抗氧化作用的天然产物，但其具体的抗氧化机制仍不清楚。本研究以犊牛睾丸支持细胞为材料，探讨褪黑素减轻支持细胞氧化应激的机制，为提高公牛生育能力奠定基础。
使用CCK8试剂盒和MDA试剂盒筛选出过氧化氢（HO）和褪黑素（MLT）的最佳处理浓度。然后，将细胞分为四组进行处理：对照组、HO处理组、MLT处理组和HO与MLT联合处理组，随后检测MDA、ROS、GSH和SOD含量。采用实时定量PCR分析和蛋白质印迹分析检测与Nrf2-Keap1通路相关的基因和蛋白质。利用免疫荧光染色分析Nrf2的变化。
研究结果表明，HO与MLT联合处理组细胞的MDA含量显著低于单独HO处理组，但与对照组无差异。与对照组相比，HO处理组细胞的ROS水平显著升高，GSH和SOD含量显著降低。与HO处理组相比，HO与MLT联合处理组细胞的ROS水平显著降低，GSH和SOD含量显著升高，但与对照组无差异。同样，MTL可以减轻HO引起的细胞Nrf2、Keap1、HO-1和NQO1表达的变化。
褪黑素激活支持细胞中的Nrf2-Keap1信号通路，提高HO-1和NQO1的表达，从而发挥其抗氧化能力。
