不同浓度左旋肉碱对人精子冷冻过程中DNA碎片与线粒体膜电位及顶体反应的影响  

Effects of L-carnitine in different concentrations on DNA fragmentation, mitochondrial membrane potential and acrosome reaction during human sperm cryopreservation

作　　者：程兰兰 谭丽[1] 冯宗刚[1] 马丽影[1] 万利静 Cheng Lanlan;Tan Li;Feng Zonggang;Ma Liying;Wan Lijing(Reproductive Center,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]郑州大学第二附属医院生殖中心,450014

出　　处：《中国实用医刊》2019年第18期7-11,共5页Chinese Journal of Practical Medicine

基　　金：河南省科技厅科技攻关项目(162102310207,142300410240).

摘　　要：目的探讨精子冷冻过程中不同浓度左旋肉碱对精子DNA碎片指数(DFI)、线粒体膜电位(MMP)、顶体反应(AR)的影响。方法随机选择2017年6月至11月于郑州大学第二附属医院就诊的患者40例,患者知情同意后进行实验。采用手淫法留取精液,所有标本充分混匀后均分为5组(每组40例)。A组:冷冻前精液组;B0组:冷冻保护剂组;B5组:添加5mmol/L左旋肉碱组;B10组:添加10mmol/L左旋肉碱组;B20组:添加20mmol/L左旋肉碱组。采用液氮蒸汽法冷冻精液,48h后复苏。采用计算机辅助精液分析系统检测各组精子的前向运动(PR)、活力;应用流式细胞仪检测各组精子的DFI、MMP以及AR。结果冷冻后精子PR、活力、MMP均低于冷冻前,精子DFI高于冷冻前;四个冷冻组间PR、活力比较差异未见统计学意义(P>0.05);DFI依次为:B10<B20<B5<B0,其中B10和B20组低于B0和B5组;MMP依次为:B0<B5<B10<B20,其中B10和B20组高于B0和B5组,差异有统计学意义(P<0.05);四个冷冻组间AR比较差异未见统计学意义(P>0.05)。结论精液冻存会造成精子的损伤,降低精子PR、活力和MMP,增加精子DFI;精子冷冻过程中添加10mmol/L或20mmol/L左旋肉碱会降低精子DFI,减轻MMP的下降,提高精子的冻存效果。因此,10mmol/L的左旋肉碱可以作为保护精子的有效成分应用到人精液冷冻中。Objective To investigate the effects of L-carnitine in different concentrations on DNA fragmentation index (DFI), mitochondrial membrane potential (MMP) and acrosome reaction (AR) during human sperm cryopreservation. Methods Forty patients were randomly selected from the June 2017 to November 2017 in the Second Affiliated Hospital of Zhengzhou University. Experiments were carried out on the patient consent. In addition, the semen samples were collected by masturbation method.All semen samples were fully mixed and divided into five groups: group A with fresh semen as control, group B0 with conventional cryoprotectant, group B5 with 5 mmol/L L-carnitine, group B10 with 10 mmol/L L-carnitine, group B20 with 20 mmol/L L-carnitine. The semen samples were frozen by liquid nitrogen vapor, and then were thawed 48h later. Computer-aided sperm analysis was used to detect the progressive motility (PR) and viability of spermatozoa before and after cryopreservation. Flow cytometry was used to detect DFI, MMP and AR in each group. Results The PR, motility and MMP after freezing were significantly lower than those before freezing. The sperm DFI was higher than that before freezing.There was no difference in the likelihoods of the PR on motility between four frozen groups (P>0.05). The sperm DFI of spermatozoa were as follows: B10<B20<B5<B0, in which B10 group and B20 group were significantly lower than the B0 group and B5 group (P<0.05). The mitochondrial membrane potentials of spermatozoa were as follows: B0<B5<B10<B20, in which B10 group and B20 group were significantly higher than the B0 group and B5 groups (P<0.05). There was no significant difference in AR between four frozen groups(P>0.05). Conclusions Sperm cryopreservation would lead to frozen injury, which can reduce PR, sperm motility, MMP Besides, sperm cryopreservation also lead to sperm DFI increasing;cryoprotectant added with 10 mmol/L L-carnitine or 20 mmol/L L-carnitine group can reduce the sperm DFI and alleviate decrease in MMP. In this way, the effects of spe

关 键 词：左旋肉碱 精子冷冻 DNA碎片 线粒体膜电位 顶体反应 

分 类 号：R714.8[医药卫生—妇产科学]