| 褪黑素的添加减轻猪精原干细胞的氧化损伤 |
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| 引用本文： | 黄诚彧,徐德全,冯哲,周玲,刘敏. 褪黑素的添加减轻猪精原干细胞的氧化损伤[J]. 畜牧兽医学报, 2021, 52(7): 1880-1890. DOI: 10.11843/j.issn.0366-6964.2021.07.011 |
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| 作者姓名： | 黄诚彧 徐德全 冯哲 周玲 刘敏 |
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| 作者单位： | 1. 华中农业大学动物医学院, 武汉 430070;2. 农业部猪遗传育种重点开放实验室, 武汉 430070;3. 农业动物遗传育种与繁殖教育部重点实验室, 武汉 430070 |
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| 基金项目： | 中央高校基本科研业务费专项资金（2662020DKPY003）；国家科技重大专项（2016ZX08006002）；湖北省农业科技创新行动项目（2018skjcx01） |
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| 摘 要： | 旨在研究褪黑素(melatonin,MT)对体外培养猪精原干细胞(spermatogonial stem cells,SSCs)的作用机制.本研究采集3头7日龄健康大白公猪睾丸,利用差速贴壁法获得SSCs.后经形态学观察、碱性磷酸酶染色、标记基因检测及免疫荧光染色鉴定后以SSCs作为试验材料,设置MT浓度梯度(0、50...
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| 关 键 词： | 精原干细胞 分离鉴定 褪黑素 氧化损伤 细胞活力 细胞凋亡 |
| 收稿时间： | 2020-09-07 |
The Addition of Melatonin Reduces the Oxidative Damage of Pig Spermatogonial Stem Cells |
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| HUANG Chengyu,XU Dequan,FENG Zhe,ZHOU Ling,LIU Min. The Addition of Melatonin Reduces the Oxidative Damage of Pig Spermatogonial Stem Cells[J]. Chinese Journal of Animal and Veterinary Sciences, 2021, 52(7): 1880-1890. DOI: 10.11843/j.issn.0366-6964.2021.07.011 |
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| Authors: | HUANG Chengyu XU Dequan FENG Zhe ZHOU Ling LIU Min |
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| Affiliation: | 1. College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China;2. Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture, Wuhan 430070, China;3. Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction (Huazhong Agricultural University) of Ministry of Education, Wuhan 430070, China |
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| Abstract: | The aim of this study was to explore the effect mechanism of melatonin(MT) on porcine spermatogonial stem cells (SSCs) in vitro culture. The SSCs in the testicles of 3 7-day-old healthy Large White pigs were isolated by differential adhesion method. After morphological observation, alkaline phosphatase staining, marker gene detection and immunofluorescence staining identification, the SSCs were used as experimental materials, the MT concentration gradient (0, 50, 250, 500, 1 000 μmol·mL-1) groups were set up to treat SSCs, each group had 3 replicates (n=3), and the blank control group was treated with 0.1% DMSO. The cell viability, reactive oxygen species (ROS), glutathione (GSH) content and apoptosis gene expression were detected after MT treatment. The results showed that:1)The isolated clonal colony cells had the growth characteristics of SSCs, which could be stained by alkaline phosphatase and the stem cell marker genes OCT4, SOX2 and SSCs marker genes NANOG, PLZF, UCHL1 were expressed; 2)After treated with more than 50 μmol·L-1 MT for 48 h, the cell viability of SSCs were significantly improved (P<0.05); 3)MT could significantly reduce the level of ROS in pig SSCs (P<0.05), and extremely significantly increase the content of intracellular GSH (P<0.01); 4)MT could significantly inhibit the expression of apoptosis proteins Bax and Caspase3 in pig SSCs (P<0.05). These results indicate that MT has the functions of removing ROS in pig SSCs, increasing GSH content, inhibiting the expression of apoptotic genes and improving cell viability, which can provide a reference for enhancing the reproductive performance of boars during breeding. |
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| Keywords: | spermatogonial stem cells isolation and identification melatonin oxidative damage cell viability apoptosis |
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