Seminal Plasma Cytokines Are Predictive of the Outcome of Boar Sperm Preservation
Seminal Plasma Cytokines Are Predictive of the Outcome of Boar Sperm Preservation
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DOI：
10.3389/fvets.2019.00436
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发表时间：
2019-12-04
影响因子：
3.2
通讯作者：
Parrilla, Inmaculada
中科院分区：
文献类型：
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作者：
Barranco, Isabel;Padilla, Lorena;Parrilla, Inmaculada
背景：猪精浆中含有丰富的细胞因子，可影响精子耐受保存的能力。目的：评估猪精浆细胞因子在猪精子储存过程中所经历的变化，无论是液体还是冷冻状态。材料和方法：在两个分离的实验中，将健康公猪和可育公猪的精液样本分成两份，其中一份进行两次离心（1500 × g， 10分钟）以收集精浆，而另一份进行商业延长（3 × 10(7)个精子/mL）并在17℃下液体储存144小时（n = 28，实验1）或使用标准0.5 mL方案进行冻融（n = 27，实验2）。使用Luminex xMAP (R)对16种细胞因子进行定量。在液体储存0、72或144小时（实验1）或冷冻前、解冻后30和150分钟（实验2）评估精子属性（casa评估总活力和进展活力；流式细胞术评估精子活力、细胞内H2O2和O-2的产生（中心点）以及活精子的脂质过氧化水平）。结果：采用贝叶斯方法进行变量选择的多元线性回归模型显示，抗炎因子tgf - β 2、tgf - β 3、IL-1Ra和IL-4以及促炎因子IL-8和IL-18可预测液体储存精液中精子活力的变化，而抗炎因子IFN-y可用于预测冷冻保存精液中所有精子属性的变化。结论：特异性的猪精浆细胞因子可调节精子在液体或冷冻状态下的结构和代谢变化。
Background: Boar seminal plasma is rich in cytokines, which could influence the capability of spermatozoa to tolerate preservation.Objectives: To evaluate the involvement of boar seminal plasma cytokines in the changes experienced by boar spermatozoa during their storage, either in liquid or frozen state.Materials and Methods: In two separated experiments, semen samples from healthy and fertile boars were split in two aliquots, one centrifuged twice (1,500 xg for 10 min) to harvest seminal plasma, whereas the other was either commercially extended (3 x 10(7) sperm/mL) and liquid-stored at 17 degrees C during 144h (n = 28, Experiment 1) or frozen-thawed using a standard 0.5 mL protocol (n = 27, Experiment 2). Sixteen cytokines were quantified using Luminex xMAP (R). Sperm attributes (CASA-evaluated total and progressive motility; flow cytometry-evaluated sperm viability, production of intracellular H2O2 and O-2(center dot)- and levels of lipid peroxidation in viable spermatozoa) were evaluated either at 0, 72, or 144 h of liquid storage (Experiment 1) or before freezing and at 30- and 150-min post-thawing (Experiment 2).Results: Multiple linear regression models, with Bayesian approach for variable selection, revealed that the anti-inflammatory TGF-beta 2, TGF-beta 3, IL-1Ra, and IL-4 and the pro-inflammatory IL-8 and IL-18, predicted changes in sperm motility for liquid-stored semen while the anti-inflammatory IFN-y was included in the models predicting changes in all sperm attributes for cryopreserved semen.Conclusion: Specific boar seminal plasma cytokines would contribute to modulate the structural and metabolic changes shown by spermatozoa during preservation, either in liquid or frozen state.