海藻糖能有效保护绵羊附睾上皮细胞免受氧化应激。

IF 1.6 3区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE Archiv Fur Tierzucht-Archives of Animal Breeding Pub Date : 2021-08-18 eCollection Date: 2021-01-01 DOI:10.5194/aab-64-335-2021
Zhaojin Luan, Xiaomei Fan, Yongchao Zhao, Huizi Song, Wei Du, Jiaoxia Xu, Zhaochen Wang, Wenguang Zhang, Jiaxin Zhang
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引用次数: 2

摘要

海藻糖是一种天然无毒的双糖,在哺乳动物中不存在,在氧化应激条件下稳定细胞膜的完整性,其机制尚不清楚。本文分析海藻糖对绵羊附睾上皮细胞(EEC)增殖的影响及其可能的机制。为了研究海藻糖对EECs的影响,从12月龄羊的睾丸中分离EECs;使用细胞计数试剂盒-8 (CCK-8)检测细胞的生长情况。通过细胞周期和凋亡检测细胞增殖,RT-PCR检测附睾分子标记物谷胱甘肽过氧化物酶5 (GPX5)和雄激素受体(AR)。接下来,用二氯-二氢-双乙酸荧光素(DCFH-DA)探针评估活性氧(ROS)含量。采用酶化学方法检测超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性,采用qRT-PCR和酶联免疫吸附试验(ELISA)检测GPX5的表达。结果表明,100 mM海藻糖显著提高了EECs的增殖能力,细胞可连续传代14次,体外GPX5和AR标记基因表达持续正常。与对照组相比,海藻糖显著提高了S期EECs的比例(P < 0.01),显著降低了EECs的凋亡率(P < 0.01)。海藻糖显著降低了EECs中ROS (P < 0.01),显著提高了CAT (P < 0.01)和GSH-Px (P < 0.05)活性。海藻糖处理的EECs中GPX5 mRNA和蛋白的表达也显著上调(P < 0.05和P < 0.01)。我们的研究表明,外源海藻糖通过提高CAT、GSH-Px活性和GPX5表达水平表现出抗氧化活性,可用于维持绵羊EECs体外长期培养的活力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Trehalose can effectively protect sheep epididymis epithelial cells from oxidative stress.

Trehalose, a naturally nontoxic disaccharide that does not exist in mammals, stabilizes cell membrane integrity under oxidative stress conditions, the mechanism of which is still unclear. Here, we analyzed the effects of trehalose on sheep epididymis epithelial cell (EEC) proliferation and its possible mechanisms. To study the effect of trehalose on EECs, EECs were isolated from testes of 12-month-old sheep; cell counting kit-8 (CCK-8) was used to measure the growth of the cells. Cell proliferation was evaluated by assaying cell cycle and apoptosis, and RT-PCR was utilized to identify the epididymal molecular markers glutathione peroxidase 5 (GPX5) and androgen receptor (AR). Next, reactive oxygen species (ROS) content was evaluated by a dichloro-dihydro-fluorescein diacetate (DCFH-DA) probe. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities were evaluated by enzyme chemistry methods, and GPX5 expression was evaluated by qRT-PCR and enzyme-linked immunosorbent assay (ELISA). The results showed that 100  mM trehalose significantly improved the proliferation potential of EECs, in which the cells could be serially passaged 14 times with continued normal GPX5 and AR marker gene expression in vitro. The trehalose can increase significantly a proportion of EECs in S phase ( P < 0.01 ) and decrease significantly the apoptotic rate of EECs ( P < 0.01 ) compared to the control. Moreover, the trehalose decreased ROS significantly ( P < 0.01 ) and increased CAT ( P < 0.01 ) and GSH-Px ( P < 0.05 ) activities significantly in EECs. GPX5 mRNA and protein expression were also significantly upregulated in trehalose-treated EECs ( P < 0.05 and P < 0.01 respectively). Our study suggested that exogenous trehalose exhibited antioxidant activity through increasing the activities of CAT, GSH-Px, and the expression level of GPX5 and could be employed to maintain vitality of sheep EECs during long-term in vitro culture.

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来源期刊
Archiv Fur Tierzucht-Archives of Animal Breeding
Archiv Fur Tierzucht-Archives of Animal Breeding 农林科学-奶制品与动物科学
CiteScore
3.20
自引率
0.00%
发文量
41
审稿时长
18-36 weeks
期刊介绍: Archives Animal Breeding is an open-access journal publishing original research papers, short communications, brief reports, and reviews by international researchers on scientific progress in farm-animal biology. The journal includes publications in quantitative and molecular genetics, genetic diversity, animal husbandry and welfare, physiology, and reproduction of livestock. It addresses researchers, teachers, stakeholders of academic and educational institutions, as well as industrial and governmental organizations in the field of animal production.
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