番茄红素、芝麻酚和黄芩素对猪精子冷冻保存效果的影响

发布时间：2018-02-28 21:34

本文关键词： 番茄红素芝麻酚黄芩素猪精子冷冻保存　出处：《西北农林科技大学》2017年硕士论文　论文类型：学位论文

【摘要】：精液冷冻保存技术是长期储存猪精子最有优势的技术,可以保存珍贵的基因物质,使精液的采集和授精在时间和空间上得以分离。与鲜精相比较,冷冻解冻后的精液具有较低的活率和生育能力,极大限制了猪精液冷冻保存技术的推广应用。本试验采用手握法采集成年杜洛克公猪精液样品,预处理后添加含有不同浓度的番茄红素、芝麻酚和黄芩素冷冻稀释液进行稀释,检测冷冻解冻后猪精子的活率、质膜完整性(低渗肿胀试验)、线粒体活性、顶体完整性、DNA完整性、脂质过氧化,以及超氧化歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性。获得如下结果:1.与对照组相比较,在冷冻稀释液中添加不同浓度的番茄红素显著提高冷冻解冻后猪精子活率、线粒体活性、质膜完整性、顶体完整性和DNA完整性(P0.05)。当番茄红素添加浓度为2.0μmol/L时,解冻后猪精子活率、线粒体活性、质膜完整性、顶体完整性较高(P0.05)。当番茄红素浓度为3.0μmol/L时,解冻后猪精子DNA完整率较高,为78.69%(P0.05)。2.冷冻稀释液中添加适宜浓度的芝麻酚,显著提高冷冻解冻后猪精子的各项品质参数(P0.05)。并且芝麻酚的最适浓度为2.0g/L(P0.05)。但是当芝麻酚的添加浓度为0.05g/L时,与对照组相比较猪精子质量参数无明显不同(P0.05)。3.冷冻稀释液中添加一定浓度的黄芩素,能对冷冻解冻过程中的猪精子具有明显的保护作用(P0.05)。当黄芩素的添加浓度为0.04g/L和0.06g/L时,获得较好的冷冻保存效果(P0.05)。4.番茄红素和芝麻酚具有协同作用,可以获得更好的冷冻保存效果。最佳联合添加浓度为0.2g/L芝麻酚+2.0μmol/L番茄红素(P0.05)。5.冷冻稀释液中单独添加一定浓度的番茄红素、芝麻酚和黄芩素,均可提高解冻后精子抗氧化酶活性(SOD、CAT、GSH-Px),并降低MDA水平(P0.05),其中,2.0μmol/L番茄红素、0.2g/L芝麻酚和0.04g/L黄芩素显著提高了三种酶的活性(P0.05)。联合添加0.2g/L芝麻酚+2.0μmol/L番茄红素和0.2g/L芝麻酚+2.5μmol/L番茄红素这两组进一步提高解冻后精子抗氧化能力(P0.05)。综上所述,在猪精子冷冻解冻过程中,这三种抗氧化剂对猪精子都有明显的保护作用。联合使用番茄红素和芝麻酚效果更佳。
[Abstract]:Semen cryopreservation is the most advantageous technique for long-term storage of pig sperm. It can preserve precious genetic material and separate semen collection and insemination in time and space. The low viability and fertility of frozen thawed semen greatly limited the popularization and application of cryopreservation technology of boar semen. In this experiment, adult Duroc boar semen samples were collected by hand holding method. After pretreatment, lycopene, sesamin and baicalein were diluted with different concentrations of lycopene, sesamin and baicalein. The motility of frozen and thawed pig spermatozoa, the integrity of plasma membrane (low osmotic swelling test, mitochondrial activity) were measured. The activities of DNA integrity, lipid peroxidation, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) were obtained as follows: 1. Different concentrations of lycopene were added to the freeze-thawed solution to significantly increase the sperm motility, mitochondrial activity, plasma membrane integrity, acrosome integrity and DNA integrity of pig spermatozoa after freezing and thawing. When lycopene was added at a concentration of 2.0 渭 mol/L, the sperm motility, mitochondrial activity, plasma membrane integrity, acrosome integrity and DNA integrity were significantly increased. The sperm motility, mitochondrial activity, plasma membrane integrity and acrosome integrity were higher after thawing. When lycopene concentration was 3.0 渭 mol/L, the DNA integrity rate of thawed porcine spermatozoa was 78.69 P0.05 路2.The appropriate concentration of sesamin was added to the frozen diluent. The quality parameters of frozen and thawed pig spermatozoa were significantly increased, and the optimum concentration of sesamin was 2.0g / L P0.05g / L, but when the concentration of sesamin was 0.05g / L, Compared with the control group, there was no significant difference in the quality parameters of porcine spermatozoa (P 0.05. 3). Adding a certain concentration of baicalin into the frozen diluent could obviously protect pig spermatozoa from freezing and thawing. When the concentration of baicalin was 0.04 g / L and 0.06 g / L, the concentration of baicalin was 0.04 g / L and 0.06 g / L, respectively. The results showed that lycopene and sesamin had synergistic effect. The best combination concentration was 0.2 g / L sesamol 2.0 渭 mol/L lycopene (P0.05). 5. Lycopene, sesamin and baicalein were added separately to the frozen diluent, and the optimum concentration of lycopene, sesamin and baicalein was determined by the addition of lycopene, sesamin and baicalin. The antioxidant enzyme activity of spermatozoa after thawing was increased and the level of MDA was decreased (P 0.05), in which the lycopene of 2.0 渭 mol/L lycopene 0.2g / L sesamin and 0.04g / L baicalin significantly increased the activity of the three enzymes (P0.05N). The combination of 0.2 g / L sesamol 2.0 渭 mol/L lycopene and 0.2 g / L sesamol was added to the thawed spermatozoa. 2. 5 渭 mol/L lycopene could further improve the antioxidant ability of thawed spermatozoa. During the freezing and thawing of porcine spermatozoa, these three antioxidants had obvious protective effects on pig spermatozoa, and the combination of lycopene and sesamin was more effective.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S828

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