Antioxidants and Oxidants in Boar Spermatozoa and Their Surrounding Environment Are Associated with AMPK Activation during Liquid Storage.
Antioxidants and Oxidants in Boar Spermatozoa and Their Surrounding Environment Are Associated with AMPK Activation during Liquid Storage.
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公猪精子及其周围环境中的抗氧化剂和氧化剂与液体储存期间 AMPK 的激活有关。
DOI：
10.3390/vetsci10030214
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发表时间：
2023-03-10
影响因子：
2.4
通讯作者：
Liu Z
中科院分区：
文献类型：
--
作者：
Li J;Zhao W;Zhu J;Ju H;Liang M;Wang S;Chen S;Ferreira-Dias G;Liu Z
猪精液17℃液态保存是猪繁殖人工授精的常规方法。在保存过程中，公猪的精子质量随着时间的推移而不断下降，这影响了人工授精后的田间结果。贮藏引起的氧化应激被认为是造成品质损失的主要原因。更好地了解精子抵抗氧化应激的机制是至关重要的。AMP激活的蛋白激酶在猪精子中被发现，在激活时对维持精子的质量和功能起到积极作用。本研究旨在探讨精子遭受氧化应激时，AMP激活的蛋白激酶是否被激活。我们的结果表明，精子质量随着保存时间的延长而下降，这与精子及其周围环境中的抗氧化剂和氧化剂水平有关，而抗氧化剂和氧化剂水平与AMP激活的蛋白激酶激活有关。用过氧化氢(一种活性氧物种)处理猪精液，证实了氧化应激在激活AMP激活的蛋白激酶中的积极作用。总而言之，在猪精液中，氧化应激促进了AMP激活的蛋白激酶的激活，这反映了氧化剂对抗氧化剂的优势。这些发现揭示了精子抵抗OS的机制，并为选择添加到精液稀释液中的抗氧化剂以获得更好的生殖效果提供了信息。激活AMP激活的蛋白激酶(AMPK)有助于提高猪精子的质量和功能，但AMPK激活猪精子的潜在机制尚不清楚。本研究旨在探讨猪精子及其周围液(SF)中抗氧化剂和氧化剂对AMPK在液体保存过程中激活的影响。收集杜洛克公猪的精液，并将其稀释到25×106/毫升的最终浓度。杜洛克公猪通常用于精液生产。在试验1中，18头公猪的25个精液样本在17℃下保存7天。试验2采用9头公猪9个精液的混合精液，每个精液用0、0.1、0.2和0.4μM/L双氧水处理，在17℃下保存3h，测定精子质量和功能、精子和SF中的抗氧化剂和抗氧化剂、胞内腺苷三磷酸/三磷酸腺苷比值以及磷酸化的腺苷三磷酸的表达水平(Thr172)。在存活率方面，精子质量随着保存时间的延长而显著下降(p<；0.05)。精子的总抗氧化能力(TAC)、丙二醛(MDA)和精子的总氧化状态(TOS)随着保存时间的延长而显著下降，精子超氧化物歧化酶样酶(SOD样)活性也有波动(P<；0.05)。细胞内AMP/ATP比值在第4天升高(p<；0.05)，随后在第6、7天降至最低值(p<；0.05)。磷酸化AMPK水平从第2天到第7天升高(p<；0.05)。相关分析表明，精子在液态保存过程中的质量与精子和SF中的抗氧化剂和氧化剂有关(p<；0.05)，与精子AMPK的磷酸化有关(p<；0.05)。与对照组相比，过氧化氢处理导致精子质量受损(p<；0.05)，抗氧化剂水平下降(SF TAC，p<；0.05；精子超氧化物歧化酶样活性，p<；0.01)，氧化剂水平增加(SF MDA，p<；0.05；细胞内ROS产生，p<；0.05)，AMP/ATP比率升高(p<；0.05)，以及磷酸化AMPK水平(p<；0.05)。这些结果表明，猪精子和SF中的抗氧化剂和氧化剂参与了AMPK在液体保存过程中的激活。
Liquid storage of boar semen at 17 °C is a conventional method used for artificial insemination in pig reproduction. During storage, boar sperm quality continuously decreases with time, which compromises the field results after artificial insemination. Oxidative stress induced by storage is considered as the main cause of the quality loss. It is crucial to have a better understanding of the mechanism of sperm resistance to oxidative stress. The AMP-activated protein kinase was found in boar spermatozoa and plays a positive role in maintaining sperm quality and functionality when activated. This study aimed to explore if AMP-activated protein kinase could be activated when spermatozoa suffer oxidative stress. Our results show that sperm quality decreased with storage time, which correlated to antioxidant and oxidant levels in spermatozoa and their surrounding environment that was correlated to the activation of the AMP-activated protein kinase. Treatment of boar semen with H2O2 (one of the reactive oxygen species) confirmed the positive role of oxidative stress in activating the AMP-activated protein kinase. Taken together, oxidative stress, reflected by the predominance of oxidants over antioxidants in boar semen, promotes activation of the AMP-activated protein kinase. The findings shed light on the mechanism of sperm resistance to OS and provide information for selecting antioxidants added into semen extender for better reproductive results. Activation of the AMP-activated protein kinase (AMPK) has been demonstrated to be beneficial for boar sperm quality and functionality, while the underlying mechanism of AMPK activation of boar spermatozoa remains obscure. This study aimed to explore the effect of antioxidants and oxidants in boar spermatozoa and their surrounding fluid (SF) on the activation of AMPK during the liquid storage. Ejaculates from Duroc boars, routinely used for semen production, were collected and diluted to a final concentration of 25 × 106/mL. In experiment 1, twenty-five semen samples from eighteen boars were stored at 17 °C for 7 days. In experiment 2, three pooled semen samples created from nine ejaculates of nine boars were used, and each sample was treated with 0, 0.1, 0.2, and 0.4 μM/L H2O2 and stored at 17 °C for 3 h. Sperm quality and functionality, antioxidants and oxidants in boar spermatozoa and SF, the intracellular AMP/ATP ratio, and the expression levels of the phosphorylated AMPK (Thr172) were determined. Sperm quality significantly decreased with storage time in terms of viability (p < 0.05). Antioxidant and oxidant levels were markedly affected with storage time, with a decline in the SF total antioxidant capacity (TAC) (p < 0.05), SF malondialdehyde (MDA) (p < 0.05), and the sperm’s total oxidant status (TOS), as well as a fluctuation in sperm superoxidase dismutase-like (SOD-like) activity (p < 0.05). The intracellular AMP/ATP ratio increased (p < 0.05) on day 4 and subsequently decreased to its lowest value on days 6 and 7 (p < 0.05). The phosphorylated AMPK levels increased from day 2 to day 7 (p < 0.05). Correlation analyses indicate that sperm quality during liquid storage was correlated to antioxidants and oxidants in spermatozoa and SF (p < 0.05), which were correlated to the phosphorylation of sperm AMPK (p < 0.05). Treatment with H2O2 induced damages in sperm quality (p < 0.05), a decline in antioxidant levels (SF TAC, p < 0.05; sperm SOD-like activity, p < 0.01), an increase in oxidant levels (SF MDA, p < 0.05; intracellular ROS production, p < 0.05), a higher AMP/ATP ratio (p < 0.05), and phosphorylated AMPK levels (p < 0.05) in comparison with the control. The results suggest that antioxidants and oxidants in boar spermatozoa and SF are involved in AMPK activation during liquid storage.
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影响因子：
3.2
作者：
通讯作者：
--
DOI：
10.3390/antiox11071279
发表时间：
2022-06-28
期刊：
Antioxidants (Basel, Switzerland)
影响因子：
--
作者：
通讯作者：
--
影响因子：
5.6
作者：
Martin-Hidalgo D;Hurtado de Llera A;Calle-Guisado V;Gonzalez-Fernandez L;Garcia-Marin L;Bragado MJ
通讯作者：
Bragado MJ
影响因子：
0.8
作者：
Dziekonska, A.;Swiader, K.;Kordan, W.
通讯作者：
Kordan, W.
DOI：
10.1073/pnas.0705070104
发表时间：
2007-07-17
影响因子：
11.1
作者：
Jaeger, Sibylle;Handschin, Christoph;Spiegelman, Bruce M.
通讯作者：
Spiegelman, Bruce M.