本文關(guān)鍵詞： 番茄紅素 芝麻酚 黃芩素 豬精子 冷凍保存　出處：《西北農(nóng)林科技大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：精液冷凍保存技術(shù)是長(zhǎng)期儲(chǔ)存豬精子最有優(yōu)勢(shì)的技術(shù),可以保存珍貴的基因物質(zhì),使精液的采集和授精在時(shí)間和空間上得以分離。與鮮精相比較,冷凍解凍后的精液具有較低的活率和生育能力,極大限制了豬精液冷凍保存技術(shù)的推廣應(yīng)用。本試驗(yàn)采用手握法采集成年杜洛克公豬精液樣品,預(yù)處理后添加含有不同濃度的番茄紅素、芝麻酚和黃芩素冷凍稀釋液進(jìn)行稀釋,檢測(cè)冷凍解凍后豬精子的活率、質(zhì)膜完整性(低滲腫脹試驗(yàn))、線粒體活性、頂體完整性、DNA完整性、脂質(zhì)過(guò)氧化,以及超氧化歧化酶(superoxide dismutase,SOD)、過(guò)氧化氫酶(catalase,CAT)和谷胱甘肽過(guò)氧化物酶(glutathione peroxidase,GSH-Px)活性。獲得如下結(jié)果:1.與對(duì)照組相比較,在冷凍稀釋液中添加不同濃度的番茄紅素顯著提高冷凍解凍后豬精子活率、線粒體活性、質(zhì)膜完整性、頂體完整性和DNA完整性(P0.05)。當(dāng)番茄紅素添加濃度為2.0μmol/L時(shí),解凍后豬精子活率、線粒體活性、質(zhì)膜完整性、頂體完整性較高(P0.05)。當(dāng)番茄紅素濃度為3.0μmol/L時(shí),解凍后豬精子DNA完整率較高,為78.69%(P0.05)。2.冷凍稀釋液中添加適宜濃度的芝麻酚,顯著提高冷凍解凍后豬精子的各項(xiàng)品質(zhì)參數(shù)(P0.05)。并且芝麻酚的最適濃度為2.0g/L(P0.05)。但是當(dāng)芝麻酚的添加濃度為0.05g/L時(shí),與對(duì)照組相比較豬精子質(zhì)量參數(shù)無(wú)明顯不同(P0.05)。3.冷凍稀釋液中添加一定濃度的黃芩素,能對(duì)冷凍解凍過(guò)程中的豬精子具有明顯的保護(hù)作用(P0.05)。當(dāng)黃芩素的添加濃度為0.04g/L和0.06g/L時(shí),獲得較好的冷凍保存效果(P0.05)。4.番茄紅素和芝麻酚具有協(xié)同作用,可以獲得更好的冷凍保存效果。最佳聯(lián)合添加濃度為0.2g/L芝麻酚+2.0μmol/L番茄紅素(P0.05)。5.冷凍稀釋液中單獨(dú)添加一定濃度的番茄紅素、芝麻酚和黃芩素,均可提高解凍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番茄紅素、0.2g/L芝麻酚和0.04g/L黃芩素顯著提高了三種酶的活性(P0.05)。聯(lián)合添加0.2g/L芝麻酚+2.0μmol/L番茄紅素和0.2g/L芝麻酚+2.5μmol/L番茄紅素這兩組進(jìn)一步提高解凍后精子抗氧化能力(P0.05)。綜上所述,在豬精子冷凍解凍過(guò)程中,這三種抗氧化劑對(duì)豬精子都有明顯的保護(hù)作用。聯(lián)合使用番茄紅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.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S828

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