番茄红素对人精子冷冻损伤的保护作用及机制研究  

Protective effect of lycopene on human spermatozoa during cryopreservation and its mechanism

作　　者：梁作文[1] 郭凯敏[1] 戴小凡[2] 刘凌云[1] 徐胜旗 赵丽晶[3] 李付彪[1] 王洪亮[1] 

机构地区：[1]吉林大学第一医院男科,吉林长春130021 [2]吉林市中心医院泌尿外科,吉林吉林132020 [3]吉林大学基础医学院病理生理学系,吉林长春130021

出　　处：《中华男科学杂志》2015年第6期521-526,共6页National Journal of Andrology

基　　金：吉林省科技发展计划项目(201015152)~~

摘　　要：目的:通过在人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可能保护作用及机制。方法:选择吉林省人类精子库捐精者的精液标本25份,每份精液一式4份,3∶1加入冷冻保护液后混匀,不含番茄红素者设为对照组(Ly0),而Ly2、Ly5、Ly10实验组混合液中分别含有2、5、10μmol/L的番茄红素,冷冻复苏精液进行常规分析,采用流式细胞术分析冻融后各组精子的凋亡率;采用硫代巴比妥酸(TBA)比色法检测精子中脂质过氧化产物丙二醛(MDA)浓度。利用JC-1标记法检测线粒体膜电位。结果:冻融后各组精子运动参数较新鲜精液参数均明显下降(P<0.01),Ly5组冻融后精子凋亡率[(25.68±4.36)%]较对照组[(33.26±4.78)%]显著下降(P<0.05);Ly5组线粒体膜电位水平[(66.18±14.23)%]显著高于对照组[(55.24±12.31)%],P<0.05;Ly2、Ly5、Ly10组的MDA水平与对照组比较,无统计学差异(P>0.05)。结论:在精液冷冻保护液中添加一定浓度的番茄红素可以减少线粒体氧化损伤,减轻活性氧对精子质膜的氧化应激损伤,从而提高精子的抗凋亡能力。Objective： To investigate the protective effect of lycopene against cryopreservation injury of post-thawing human sperm and its mechanism. Methods： Semen samples were collected from 25 volunteers,each sample equally divided into four parts to be cryopreserved with cryoprotectant only（ Ly0 control） or cryoprotectant ＋ lycopene at the concentrations of 2（ Ly2）,5（ Ly5）,and 10 μmol / L（ Ly10）,respectively. Before and after thawing,the semen samples were subjected to computer-assisted semen analysis（ CASA） for sperm kinematics,flow cytometry for sperm apoptosis,thiobarbituric acid assay for malondialdehyde（ MDA） concentration,and JC-1 fluorescent staining for the sperm mitochondrial membrane potential（ MMP）. Results： After cryopreservation,sperm motility was markedly decreased in all the groups（ P 〈 0. 01）. The rate of sperm apoptosis was significantly lower in the Ly5 group than in the Ly0 control（[25. 68 ± 4. 36]% vs [33. 26 ± 4. 78]%,P 〈 0. 05）,while sperm MMP remarkably higher in the former than in the latter（[66. 18 ± 14. 23]% vs [55. 24 ± 12. 31]%,P 〈 0. 05）. The Ly2,Ly5 and Ly10 groups showed no statistically significance differences in the MDA level from the Ly0 control（ P 〉 0. 05）. Conclusion： Addition of lycopene at a proper concentration to cryoprotectant may reduce oxidative damage to sperm mitochondria in the freezing-thawing process,attenuate oxidative stress injury induced by reactive oxygen species to sperm plasma membrane,and improve the anti-apoptosis ability of sperm.

关 键 词：番茄红素 精子 冷冻保护 氧化应激损伤 凋亡 线粒体膜电位 

分 类 号：R698.2[医药卫生—泌尿科学]