异柠檬酸在精子冻融中保护作用的初探  

Preliminary study on the protective effect of isocitric acid in sperm cryopreservation

作　　者：赵凯[1] 陈伟[2] 徐爱明[1] 张建中[1] 华艺博 王增军[1] 

机构地区：[1]南京医科大学第一附属医院泌尿外科,江苏南京210029 [2]南京医科大学附属南京医院泌尿外科,江苏南京210006

出　　处：《南京医科大学学报（自然科学版）》2017年第2期165-168,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金项目(81270685;81402104)

摘　　要：目的:探讨异柠檬酸在精子冻融中的保护作用和机制。方法:选取16例健康生育男性的精液样本,分析精液常规参数后,将每份精液1式5份,1∶1加入冷冻保护剂后混匀,不含异柠檬酸的为阴性对照组,实验组分别添加1.25、2.50、3.75、5.00 mmol/L浓度的异柠檬酸,冷冻复苏后进行精液常规分析。同时另选取9例健康生育男性的精液样本,采用荧光染料5,5′,6,6′四氯-1和1′,3,3′四乙基苯并咪唑基羰化青碘(5,5′,6,6′-tetrachloro-1,1′,3,3'′-tetraethyl-imidacarbo-cyanine iodide,JC-1)单标法检测复苏后精子细胞的线粒体膜电位。结果:复苏后G3组前向运动精子百分率(PR)升高(P<0.01),G2组PR升高(P<0.05),G3组精子浓度上升(P<0.05);G3组精子复苏后线粒体膜电位较对照组增大(P<0.05)。结论:精液冷冻保护剂中添加适宜浓度的异柠檬酸可以保护冷冻复苏后精子线粒体功能,提高复苏后精子活力和浓度。Objective：To explore the potential protective effects and mechanism of isocitric acid in post-thawing sperm. Methods：After routine semen analysis,semen samples from 16 fertile males were selected. Every sample was equally divided into five groups.One group was mixed with routine protectant as negative control,and the other groups were mixed with modified protectant containing different concentration of isocitric acid. After and before thawing,the sperm samples were subjected to CASA for analysis and flow cytometry for mitochondrial transmembrane potential,respectively. Results：The percentage of forward movement sperm in G2 and G3 group was higher than that of the control group（P〈0.05 and P〈0.01）. Sperm concentration of G3 group also increased（P〈0.05）.The mitochondrial membrane potential in the G3 group was higher than that of the control group（P〈0.05）.Conclusion：Adding a certain concentration of citric acid in the cryoprotectant can reduce the damage of the mitochondria during the freezing and thawing.

关 键 词：异柠檬酸 前向运动精子 线粒体膜电位 精液 冷冻保护剂 

分 类 号：R392.69[医药卫生—免疫学]