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谷胱甘肽对关中奶山羊精子冷冻保存过程中精子质量的影响

Effect of Glutathione on Sperm Quality in Guanzhong Dairy Goat Sperm During Cryopreservation.

作者信息

Zou Jiahao, Wei Lixuan, Li Dexian, Zhang Yongtao, Wang Guang, Zhang Lei, Cao Ping, Yang Shaohua, Li Guang

机构信息

College of Animal Science and Technology, Northwest A&F University, Yangling, China.

Xi'an Dairy Cow Breeding Center, Xi'an Agriculture and Rural Affairs Bureau, Xi'an, China.

出版信息

Front Vet Sci. 2021 Nov 18;8:771440. doi: 10.3389/fvets.2021.771440. eCollection 2021.

Abstract

In the process of cryopreservation of dairy goat semen, it will face many threats such as oxidative damage, which will affect the motility and plasma membrane function of sperm. As an endogenous antioxidant in animals, glutathione (GSH) can significantly improve the quality of thawed sperm when added to the frozen diluent of semen of pigs and cattle. In this study, different concentration gradients of GSH [0 mmol/L (control), 1, 2, 3, 4 mmol/L] were added to the frozen diluent of Guanzhong dairy goat semen. By detecting the sperm motility parameters, acrosome intact rate and plasma membrane intact rate after thawing, the effect of GSH on the cryopreservation of dairy goat semen was explored. Sperm motility parameters were measured with the computer-aided sperm analysis (CASA) system (total power, TM; forward power, PM; linearity, LIN; average path speed, VAP; straight line speed, VSL; curve speed, VCL; beat cross frequency, BCF). The sperm acrosome integrity rate after thawing was detected by a specific fluorescent probe (isothiocyanate-labeled peanut agglutinin, FITC-PNA), and the sperm plasma membrane integrity rate after thawing was detected by the hypotonic sperm swelling (HOST) method. Reactive oxygen species (ROS) kit, malondialdehyde (MDA) kit, superoxide dismutase (SOD) kit, glutathione peroxidase (GSH-PX) kit were used to detect various antioxidant indicators of thawed sperm. fertilization experiment was used to verify the effect of adding glutathione on sperm fertilization and embryo development. The results showed that when the concentration of glutathione was 2 mmol/l, the sperm viability, plasma membrane intact rate, and acrosome intact rate were the highest after thawing, reaching 62.14, 37.62, and 70.87% respectively, and they were all significantly higher. In terms of antioxidant indexes; the values of SOD and GSH-PX were 212.60 U/ml and 125.04 U/L, respectively, which were significantly higher than those of the control group; The values of ROS and MDA were 363.05 U/ml and 7.02 nmol/L, respectively, which were significantly lower than the control group. The addition of 2 mmol/L glutathione significantly improves the fertilization ability of sperm. In short, adding 2 mmol/l glutathione to the semen diluent can improve the quality of frozen Guanzhong dairy goat sperm.

摘要

在关中奶山羊精液冷冻保存过程中,会面临氧化损伤等诸多威胁,这会影响精子的活力和质膜功能。谷胱甘肽(GSH)作为动物体内的一种内源性抗氧化剂,添加到猪和牛精液的冷冻稀释液中时,能显著提高解冻后精子的质量。在本研究中,将不同浓度梯度的GSH[0 mmol/L(对照组)、1、2、3、4 mmol/L]添加到关中奶山羊精液的冷冻稀释液中。通过检测解冻后的精子活力参数、顶体完整率和质膜完整率,探讨GSH对奶山羊精液冷冻保存的影响。精子活力参数采用计算机辅助精子分析(CASA)系统进行测定(总活力,TM;前进活力,PM;直线性,LIN;平均路径速度,VAP;直线速度,VSL;曲线速度,VCL;鞭打交叉频率,BCF)。解冻后的精子顶体完整率采用特异性荧光探针(异硫氰酸酯标记的花生凝集素,FITC-PNA)进行检测,解冻后的精子质膜完整率采用低渗精子肿胀(HOST)法进行检测。采用活性氧(ROS)试剂盒、丙二醛(MDA)试剂盒、超氧化物歧化酶(SOD)试剂盒、谷胱甘肽过氧化物酶(GSH-PX)试剂盒检测解冻后精子的各项抗氧化指标。通过受精试验验证添加谷胱甘肽对精子受精和胚胎发育的影响。结果表明,当谷胱甘肽浓度为2 mmol/L时,解冻后精子的活力、质膜完整率和顶体完整率最高,分别达到62.14%、37.62%和70.87%,且均显著高于对照组。在抗氧化指标方面,SOD和GSH-PX的值分别为212.60 U/ml和125.04 U/L,显著高于对照组;ROS和MDA的值分别为363.05 U/ml和7.02 nmol/L,显著低于对照组。添加2 mmol/L谷胱甘肽能显著提高精子的受精能力。总之,在精液稀释液中添加2 mmol/L谷胱甘肽可提高冷冻保存的关中奶山羊精子的质量。

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