褪黑素的添加减轻猪精原干细胞的氧化损伤

doi:10.11843/j.issn.0366-6964.2021.07.011

Abstract

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.

Key words: spermatogonial stem cells, isolation and identification, melatonin, oxidative damage, cell viability, apoptosis

CLC Number:

S828.3

HUANG Chengyu, XU Dequan, FENG Zhe, ZHOU Ling, LIU Min. The Addition of Melatonin Reduces the Oxidative Damage of Pig Spermatogonial Stem Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(7): 1880-1890.

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The Addition of Melatonin Reduces the Oxidative Damage of Pig Spermatogonial Stem Cells

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