【摘要】：兔精子在冷凍保存的過程中,由于遭受冷休克而產(chǎn)生過量的對精子有害的氧自由基,致使精子容易發(fā)生脂質(zhì)過氧化反應(yīng),從而使精子的質(zhì)膜遭到破壞,導(dǎo)致解凍后的精子品質(zhì)降低、人工授精率、分娩率降低,限制了兔人工授精技術(shù)的推廣運用。本試驗擬在兔精子冷凍保存液TCG中添加不同濃度的褪黑色素,初步探討褪黑色素對兔精子冷凍保存的影響,利用光學(xué)顯微鏡觀察精子活率,且利用SYBR14/PI、FITC-PNA、JC-1、BODIPY581/591C11熒光染料檢測解凍后的精子的質(zhì)膜完整率、頂體完整率、線粒體膜電位變化及脂質(zhì)過氧化反應(yīng),此外,還對褪黑色素處理的凍存后的精子進行凋亡的檢測,研究獲得以下結(jié)果:1.在兔精子冷凍保存稀釋液中,添加不同濃度的褪黑色素,與對照組相比,可以顯著提高解凍后的兔精子的活率、質(zhì)膜完整率、頂體完整率、線粒體膜電位變化及降低了精子的脂質(zhì)過氧化反應(yīng)(P0.05),且褪黑色素的推薦添加濃度為0.1或1mM,當(dāng)添加濃度為5mM時,效果不顯著(P0.05)。2.在精子冷凍稀釋液中,添加褪黑色素可以減緩精子因在冷凍-解凍過程中遭受損傷而引起的凋亡現(xiàn)象。試驗發(fā)現(xiàn),當(dāng)在稀釋液中添加1mM的褪黑色素時,精子的凋亡現(xiàn)象可以得到緩解。因此,在兔精子冷凍保存中,添加0.1mM的褪黑色素可能通過減緩精子的凋亡,從而提高解凍后的精子品質(zhì)。
[Abstract]:During the cryopreservation of rabbit spermatozoa, excessive oxygen free radicals harmful to sperm are produced due to cold shock, which makes the sperm prone to lipid peroxidation, thus destroying the plasma membrane of the sperm. After thawing, the sperm quality, artificial insemination rate and delivery rate decreased, which limited the application of artificial insemination technology in rabbits. In this experiment, different concentrations of melatonin were added to rabbit sperm cryopreservation solution (TCG). The effect of melatonin on rabbit sperm cryopreservation was studied. The sperm motility was observed by optical microscope and SYBR14/PI,FITC-PNA,JC-1, was used. The plasma membrane integrity rate, acrosome integrity rate, mitochondrial membrane potential change and lipid peroxidation of thawed spermatozoa were detected by BODIPY581/591C11 fluorescent dye. In addition, apoptosis of frozen spermatozoa treated with melatonin was also detected. The results are as follows: 1. Compared with the control group, the sperm motility, plasma membrane integrity and acrosome integrity of rabbit spermatozoa were significantly increased by adding melatonin at different concentrations in the rabbit sperm cryopreservation diluent. The change of mitochondrial membrane potential and the decrease of lipid peroxidation of spermatozoa (P0.05), and the recommended concentration of melatonin was 0.1 or 1 mm, but the effect was not significant when the concentration of 5mM was added (P0.05). The addition of melatonin in semen cryopreservation diluents can attenuate the apoptosis of spermatozoa caused by injury during freezing and thawing. It was found that the apoptosis of spermatozoa could be alleviated when 1mM melatonin was added to the diluent. Therefore, in cryopreservation of rabbit spermatozoa, melatonin added with 0.1mM may improve the quality of thawed spermatozoa by slowing down the apoptosis of spermatozoa.
【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】：S829.1

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