维生素E类似物对兔精子耐冻性的影响

发布时间：2018-03-28 17:06

本文选题：兔精液　切入点：冷冻保存　出处：《西北农林科技大学》2015年硕士论文

【摘要】：近年来,随着经济的发展,人们对生活各方面的质量和要求也越来愈高,首先就表现在饮食方面。我国是农业大国,畜牧业是其中重要的组成部分,然而,传统的高脂、高能的饮食习惯对我们的健康造成了很大的影响。而兔肉具有低脂肪、低胆固醇、低热量的特点,能满足人们的健康饮食。此外,我国人多地少,存在粮食供应与需求的矛盾,猪、禽等都属于耗粮型畜禽,与人争粮的矛盾日益突出,而兔属于节粮型家畜,可以有效的利用饲草资源,以节省粮食。因此,大力发展养兔业对促进我国经济发展有重要作用。但是,公兔具有夏季不育的现象,在高温热应激下,会导致精液量下降,精子密度减少,品质下降等。因此,需要将兔精液冷冻保存,以解决夏季无法配种的问题。然而,目前兔精液冷冻保存的效果并不理想,因此,尚未被广泛的应用于生产实践中。为了进一步改善冷冻保存后兔精液品质,本试验从抗氧化剂方面对稀释液进行改进,通过向冷冻保存液中添加水溶性维生素E类似物(Trolox),来研究其对兔精液冷冻保存后精子品质的影响。试验以冷冻-解冻后精子活力、质膜完整性、顶体完整性、DNA完整性、脂质过氧化水平以及线粒体活性等作为评价指标,利用碘化丙锭(PI)、SYBR-14、异硫氰酸荧光素标记的花生凝集素(FITC-PNA)、吖啶橙(AO)、BODIPY-C11荧光探针、5,5',6,6'-四氯-1,1',3,3'-四乙基苯丙咪唑碘化羧花青(JC-l)等对精子的质量进行评估。本文所得结果如下:1.兔精液冷冻保存液中添加Trolox,解冻后各试验组的精子活力、顶体完整性、线粒体活性均显著高于对照组(P㩳0.05),脂质过氧化水平均显著低于对照组(P㩳0.05),添加浓度为200μM时,质膜完整性显著高于对照组(P㩳0.05)。2.比较不同浓度的Trolox对精液品质的影响,解冻后添加浓度为200μM的试验组精子活力、顶体完整性、线粒体活性均显著高于浓度为100μM和250μM的试验组(P㩳0.05),浓度为100μM和200μM的试验组其脂质过氧化水平显著低于250μM的试验组(P㩳0.05)。本试验结果表明维生素E类似物能增加兔精子的耐冻性,并且维生素E类似物最适添加浓度为200μM。
[Abstract]:In recent years, with the development of economy, people's quality and demand for all aspects of life have become higher and higher, first of all, in the aspect of diet. China is a big agricultural country, animal husbandry is an important part of it, however, the traditional high fat, High energy eating habits have a great impact on our health. Rabbit meat has the characteristics of low fat, low cholesterol and low calorie, which can satisfy people's healthy diet. In addition, there is a contradiction between food supply and demand. Pigs and poultry are all food consuming animals and poultry, and the contradiction of competing with people for grain is increasingly prominent, while rabbits are grain saving livestock, which can effectively use forage resources in order to save grain. The development of rabbit breeding industry plays an important role in promoting the economic development of our country. However, the male rabbit is sterile in summer. Under the high temperature heat stress, it will lead to the drop of spermatozoa, the decrease of sperm density, the decline of quality, etc. It is necessary to cryopreservation rabbit semen in order to solve the problem of inbreeding in summer. However, at present, the effect of cryopreservation of rabbit semen is not satisfactory, so, In order to further improve the quality of rabbit semen after cryopreservation, the diluent was improved in terms of antioxidants. To study the effect of water soluble vitamin E analogue troloxane on the sperm quality after cryopreservation of rabbit semen, the sperm motility, plasma membrane integrity, acrosomal integrity and DNA integrity were studied after freezing and thawing. Lipid peroxidation level and mitochondrial activity were used as evaluation indexes. The quality of spermatozoa was evaluated by using the fluorescent probe PIH4 SYBR-14, fluorescein isothiocyanate labeled peanut agglutinin (FITC-PNAA), acridine orange (Acridine Orange AOA) BODIPY-C11 as a fluorescence probe of 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 4, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 5, 4, 5, 5, 5, 4, 5, 4, 4, 4, 4, Troloxx was added to the semen cryopreservation solution, and the sperm motility of the thawed groups was determined. Acrosomal integrity and mitochondrial activity were significantly higher than those of the control group. The level of lipid peroxidation was significantly lower than that of control group. When the concentration was 200 渭 M, the integrity of plasma membrane was significantly higher than that of the control group. After thawing, the sperm motility, acrosome integrity and mitochondrial activity were significantly higher in the thawed group than those in the 100渭 M and 250渭 M groups, and the sperm motility, acrosome integrity and mitochondrial activity in the thawed group were significantly higher than those in the control group (100 渭 M and 250 渭 M, respectively). The levels of lipid peroxidation in 100 渭 M and 200 渭 M groups were significantly lower than those in 250 渭 M group. The results showed that vitamin E analogue could increase the freezing resistance of rabbit spermatozoa, and the optimum concentration of vitamin E analogue was 200 渭 M.
【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】：2015
【分类号】：S829.1

【参考文献】