番茄红素对精子冷冻损伤的保护作用及机制的初步研究

发布时间：2018-03-26 10:43

本文选题：番茄红素　切入点：冷冻保存　出处：《吉林大学》2014年硕士论文

【摘要】：目的：通过在精子冷冻保护液中添加不同浓度的番茄红素，研究番茄红素对精子冷冻损伤的保护作用及机制。方法：选择2013年9月至2014年3月来自吉林大学第一医院吉林省人类精子库捐精者的精液标本，共计25份；采用计算机辅助精液常规参数分析（CASA）冻前和冻后精液标本的精子前向运动力(a+b)、精子活动率(a+b+c)；采用硫代巴比妥酸(TBA)比色法，测定冻后精子丙二醛（MDA）的含量，作为精子膜脂质过氧化反应程度的指标；应用原位末端转移酶标记技术（TUNEL）法通过激光共聚焦显微镜观察精子DNA损伤情况，测定阳性细胞比率作为反映冻后精子DNA完整性的指标。利用JC-1标记法通过激光共聚焦显微镜检测线粒体膜电位水平，作为反映冻后精子线粒体损伤的指标。结果：精液冷冻保存后各组的精子前向运动力和活力均比新鲜组精液的参数下降（P＜0.05)，，冷冻后添加不同浓度的番茄红素其线粒体膜电位水平均明显高于对照组（P＜0.05)。添加5mol/L番茄红素组其TUNEL阳性细胞比率显著低于对照组（P＜0.05)。其余实验组的检测指标与对照组相比无明显统计学意义（P0.05)。结论：冷冻保存能够使精子的移动力、活力发生显著下降，在精液中添加番茄红素可以减少线粒体氧化损伤，精子冷冻保护液中添加适当浓度的番茄红素可以保护精子DNA的完整性。番茄红素作为抗氧化剂，以适当的浓度添加在精液冷冻保护液中可以明显改善精子质量。
[Abstract]:Objective:. The protective effect and mechanism of lycopene on sperm cryopreservation injury were studied by adding lycopene with different concentrations. Methods: from September 2013 to March 2014, 25 semen samples were collected from donors from Jilin Province Human sperm Bank, first Hospital of Jilin University. The sperm motility and sperm motility were analyzed by computer aided semen routine parameters before and after freezing, and the content of malondialdehyde (MDAs) in frozen sperm was determined by thiobarbituric acid TBA colorimetry. The degree of lipid peroxidation of sperm membrane was determined by in situ terminal transferase labeling (Tunel) technique, and the DNA damage was observed by confocal laser microscopy. The ratio of positive cells was used as an index to reflect the integrity of DNA in frozen spermatozoa and the mitochondrial membrane potential level was detected by laser confocal microscope with JC-1 labeling method as an index to reflect the damage of sperm mitochondria after freezing. Results:. The sperm forward motility and motility of each group after cryopreservation were significantly lower than that of fresh semen (P < 0.05). The mitochondrial membrane potential level of lycopene added with different concentrations of lycopene after freezing was significantly higher than that of control group (P < 0.05). The percentage of TUNEL positive cells in lycopene group was significantly lower than that in control group (P < 0.05). Conclusion: cryopreservation can significantly decrease the motility and motility of spermatozoa, and the addition of lycopene in semen can reduce the oxidative damage of mitochondria. The integrity of sperm DNA could be protected by the addition of lycopene in the cryopreservation solution, and the quality of spermatozoa could be improved by adding lycopene as an antioxidant.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R698.2

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