本文選題：豬 + 精子��； 參考：《西北農(nóng)林科技大學(xué)》2015年碩士論文

【摘要】：為了提高養(yǎng)豬的效益,豬的人工授精技術(shù)得到了廣泛的應(yīng)用。但是由于豬精液在冷凍保存過程中容易受到低溫打擊,造成解凍后豬精子的活率不甚理想,從而影響繁殖率,因此找到一些冷凍保護(hù)劑成為了亟需解決的問題。豬精子質(zhì)膜中含有較多的不飽和脂肪酸,冷凍保存及冷凍解凍過程中容易被氧化,從而破壞了精子質(zhì)膜完整性。而竹葉黃酮、異抗壞血酸鈉和丁基羥基茴香醚作為食品添加劑中幾種常見的高效抗氧化劑,在食品抗氧化方面的研究較多,但是其對豬精液冷凍保存的影響還未見相關(guān)報道。本試驗以竹葉黃酮、異抗壞血酸鈉和丁基羥基茴香醚為冷凍保護(hù)劑,分別以不同的濃度加入豬精液冷凍稀釋液,通過顯微鏡觀察、碘化丙啶(PI)和羅丹明123聯(lián)合染色、低滲腫脹實驗(HOST)以及異硫氰酸熒光素連接的花生凝集素(PITC-PNA)分別檢測了解凍后豬精子的活力、活率、線粒體膜完整性、質(zhì)膜完整性和頂體完整性,評價幾種抗氧化劑對豬精液冷凍保存的保護(hù)作用。同時,還利用酶活性檢測試劑盒檢測了冷凍-解凍后豬精子的丙二醛(MDA)水平、超氧化物歧化酶(SOD)、谷胱甘肽過氧化物酶(GSH-Px)等酶的活性,得出了以下幾個結(jié)論:1.將不同濃度的竹葉黃酮添加到豬精液冷凍稀釋液中,可以顯著提高冷凍解凍后豬精子的活力、活率、線粒體膜完整率、質(zhì)膜完整率和頂體完整率(P0.05)。當(dāng)竹葉黃酮的濃度為5 mg/mL時,冷凍解凍后豬精子的活力、活率、線粒體膜完整率、質(zhì)膜完整率和頂體完整率最高。2.在豬精液冷凍稀釋液中添加不同濃度的異抗壞血酸鈉,可以顯著提高冷凍解凍后豬精子的活力、活率、線粒體膜完整率、質(zhì)膜完整率和頂體完整率(P0.05)。當(dāng)異抗壞血酸鈉的濃度為6 mg/mL時,冷凍解凍后豬精子的活力、活率、線粒體膜完整率、質(zhì)膜完整率和頂體完整率最高。3.在豬精液冷凍稀釋液中添加不同濃度的丁基羥基茴香醚,可以顯著提高冷凍解凍后豬精子的活力、活率、線粒體膜完整率、質(zhì)膜完整率和頂體完整率(P0.05)。當(dāng)丁基羥基茴香醚的濃度為0.2 mg/mL時,冷凍解凍后豬精子的活力、活率、質(zhì)膜完整率和頂體完整率最高,而當(dāng)丁基羥基茴香醚的濃度為0.15 mg/mL時,線粒體膜完整率最高。4.在豬精液冷凍稀釋液中添加2 mg/mL~6 mg/mL的竹葉黃酮,與對照組相比,MDA水平的降低、SOD酶活力和GSH-Px酶活力的升高具有顯著性差異(P0.05)。當(dāng)竹葉黃酮濃度為4 mg/mL和5 mg/mL時,MDA水平降低最多;濃度為5 mg/mL時,SOD酶活力效果最好;濃度為4 mg/mL時,GSH-Px酶活力最佳。冷凍稀釋液中添加6 mg/mL異抗壞血酸鈉時,MDA水平下降最明顯;添加8 mg/mL時,SOD酶活力最高;而異抗壞血酸鈉濃度為6 mg/mL~10 mg/mL時,GSH-Px酶活力比其他組差異顯著。在冷凍稀釋液中添加不同濃度的丁基羥基茴香醚時,MDA水平、SOD酶活力和GSH-Px酶活力比對照組差異顯著(P0.05),且濃度為0.2 mg/mL時,效果均為最佳。5.兩種抗氧化劑聯(lián)合作用能顯著提高解凍后豬精子的活力、活率和膜完整性(P0.05)。其中5 mg/mL竹葉黃酮和6 mg/mL的異抗壞血酸鈉配伍作用后的效果最好。
[Abstract]:In order to improve the benefit of pig raising, pig artificial insemination technology has been widely used. However, because the pig semen is vulnerable to cryopreservation during cryopreservation, the survival rate of pig sperm is not ideal after thawing, thus affecting the reproductive rate. Therefore, finding some cryopreservation agents has become an urgent problem to be solved. More unsaturated fatty acids, frozen storage and freezing thawing are easily oxidized, thus destroying the integrity of sperm plasma membrane. However, bamboo leaf flavonoids, sodium isoascorbate and butyl hydroxy anisole are used as some common high effective antioxidants in food additives, but they have more research on the antioxidation of food products, but they have a good effect on pig sperm. The effect of liquid cryopreservation has not been reported. In this experiment, the freeze protectant of bamboo leaf flavone, sodium isoascorbate and butyl hydroxy anisole was added to the cryopreservation agent at different concentrations, respectively, by microscope observation, PI and Luo Danming 123 combined dyeing, hypotonic swelling test (HOST) and isothiocyanate. Peanut agglutinin (PITC-PNA) was used to detect the viability, viability, membrane integrity, membrane integrity and acrosome integrity of the porcine sperm after thawing, respectively, to evaluate the protective effect of several antioxidants on the cryopreservation of porcine semen. At the same time, the enzyme activity detection kit was used to detect the malondialdehyde (MD) of frozen thawing sperm. A) levels, the activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and other enzymes, and the following conclusions were obtained: 1. adding different concentrations of bamboo leaf flavonoids to the cryopreservation diluents of porcine semen can significantly improve the viability, survival rate, mitochondrial membrane integrity, membrane integrity and acrosome integrity after freezing and thawing. P0.05) when the concentration of the bamboo leaf flavonoids was 5 mg/mL, the vitality, the survival rate, the membrane integrity of the mitochondria, the integrity of the plasma membrane and the highest acrosome rate of.2. after the freezing and thawing of the pig sperm were added with different concentrations of sodium isoascorbate in the cryopreservation solution of the pig's semen, which could significantly improve the viability, the survival rate and the complete mitochondrial membrane after the freezing thawing. Rate, plasma membrane integrity and acrosome integrity (P0.05). When the concentration of sodium isoascorbate is 6 mg/mL, the vitality, viability, membrane integrity, integrity of plasma membrane and the highest acrosome integrity after freezing thawing can be added to the freezing thawing solution of the pig semen with different concentrations of butyl hydroxy anisole, which can significantly improve the freezing thawing. The viability, survival rate, mitochondrial membrane integrity, membrane integrity and acrosome integrity (P0.05). When the concentration of butyl hydroxy anisole was 0.2 mg/mL, the viability, viability, plasma membrane integrity and acrosome integrity were the highest after freezing thawing, and the mitochondrial membrane integrity was the highest when the concentration of Ding Jiqiang anisole was 0.15 mg/mL High.4. added 2 mg/mL~6 mg/mL of bamboo leaf flavonoids in the frozen semen of pigs. Compared with the control group, the MDA level decreased, the activity of SOD enzyme and the activity of GSH-Px enzyme increased significantly (P0.05). When the concentration of the bamboo leaf flavonoids was 4 mg/mL and 5 mg/mL, the MDA level decreased most; when the concentration was 5 mg/mL, the activity of the SOD enzyme was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration was the best; concentration At 4 mg/mL, GSH-Px enzyme activity was the best. When 6 mg/mL isoascorbate was added to the frozen diluent, the level of MDA decreased most obviously; when adding 8 mg/mL, the activity of SOD enzyme was the highest; and when the concentration of sodium ascorbate was 6 mg/mL~10 mg/mL, the activity of GSH-Px enzyme was significantly different. When aromatic ether, MDA level, SOD enzyme activity and GSH-Px enzyme activity were significantly different than those of the control group (P0.05). When the concentration was 0.2 mg/mL, the combined effect of the best.5. and two kinds of antioxidants could significantly improve the viability, viability and membrane integrity (P0.05) of the porcine sperm after thawing. The compatibility of 5 mg/mL bamboo leaf flavonoids and 6 mg/mL isoascorbate sodium. The latter is the best.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S828

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 吳永明;夏欣一;黃宇烽;;精子DNA完整性檢測技術(shù)研究進(jìn)展[J];中華男科學(xué)雜志;2006年08期

2 趙憲林;昝林森;胡建宏;桂林生;郝瑞杰;;維生素E對秦川牛冷凍精液的抗氧化保護(hù)作用[J];畜牧獸醫(yī)學(xué)報;2010年01期

3 張英;天然功能性竹葉提取物——竹葉黃酮[J];中國食品添加劑;2002年03期

4 周定杰;靳鐳;;熒光染色技術(shù)在精子檢測中的應(yīng)用[J];中國優(yōu)生與遺傳雜志;2007年07期

，

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