Chen Yanru, Tang Ying, Liu Bojing, Wang Junying, Wang Hongzhang, Li Bo, Liu Shicheng, Adeniran Samson O, Zheng Peng
College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, China.
Biotechnology Unit, Department of Biological Sciences, College of Basic and Applied Sciences, Mountain Top University, Ibafo, Ogun State, Nigeria.
Theriogenology. 2025 Feb;233:112-122. doi: 10.1016/j.theriogenology.2024.11.021. Epub 2024 Nov 27.
Inflammation responses and oxidative stress adversely affect testicular function, reducing fertility. Melatonin exhibits anti-inflammatory and antioxidant properties. However, the molecular mechanism by which melatonin alleviates inflammation and oxidative stress in Leydig cells of the Min pig testis remains unclear. To investigate this, primary Leydig cells were isolated from 7-day-ld Min pigs' testes and treated with LPS, HO and melatonin, respectively. The results showed that co-treatment with melatonin and LPS significantly decreased the expression of TLR4, NF-κB, IL-6 and IL-1β compared to LPS group. Co-treatment with melatonin and HO significantly mitigated reactive oxygen species and malondialdehyde levels. Melatonin also enhanced glutathione and superoxide dismutase levels and upregulated the mRNA expression levels of Nrf2, Keap1, HO-1 and NQO1. In the co-treatment group of melatonin, LPS, and SIRT1 inhibitor, the secretion of IL-6 and IL-1β and the mRNA expression levels of TLR4 and NF-κB were elevated significantly compared to the control group and the melatonin-LPS co-treatment group. In the combined treatment group of melatonin, HO and a SIRT1 inhibitor, ROS levels increased significantly, while the expression of Nrf2, Keap1, HO-1 and NQO1 were decreased significantly compared to the control group and the melatonin-HO co-treatment group. Furthermore, mRNA expression levels of testosterone synthesis-related genes StAR, CYP11A1, 3β-HSD, CYP17A1 and 17β-HSD significantly decreased following HO treatment, which was alleviated by co-treatment with HO and melatonin, but not by the addition of SIRT1 inhibitor. In conclusion, melatonin exhibits the capability to ameliorate inflammatory responses, oxidative stress and testosterone secretion in Leydig cells via the SIRT1 pathway.
炎症反应和氧化应激会对睾丸功能产生不利影响，降低生育能力。褪黑素具有抗炎和抗氧化特性。然而，褪黑素减轻小型猪睾丸间质细胞炎症和氧化应激的分子机制尚不清楚。为了研究这一点，从7日龄小型猪的睾丸中分离出原代间质细胞，并分别用脂多糖（LPS）、过氧化氢（HO）和褪黑素进行处理。结果表明，与LPS组相比，褪黑素与LPS共同处理显著降低了TLR4、NF-κB、IL-6和IL-1β的表达。褪黑素与HO共同处理显著减轻了活性氧和丙二醛水平。褪黑素还提高了谷胱甘肽和超氧化物歧化酶水平，并上调了Nrf2、Keap1、HO-1和NQO1的mRNA表达水平。在褪黑素、LPS和SIRT1抑制剂的共同处理组中，与对照组和褪黑素-LPS共同处理组相比，IL-6和IL-1β的分泌以及TLR4和NF-κB的mRNA表达水平显著升高。在褪黑素、HO和SIRT1抑制剂的联合处理组中，与对照组和褪黑素-HO共同处理组相比，活性氧水平显著升高，而Nrf2、Keap1、HO-1和NQO1的表达显著降低。此外，HO处理后，睾酮合成相关基因StAR、CYP11A1、3β-HSD、CYP17A1和17β-HSD的mRNA表达水平显著下降，HO与褪黑素共同处理可缓解这一现象，但添加SIRT1抑制剂则不能。总之，褪黑素能够通过SIRT1途径改善间质细胞中的炎症反应、氧化应激和睾酮分泌。
