论文部分内容阅读
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 protective effect and mechanism of isocitrate in sperm freezing and thawing. Methods: Semen samples of 16 healthy male fertility males were selected. After the routine parameters of semen were analyzed, 1 part of seminal fluid was divided into 5 parts, 1: 1 was added into the cryoprotectant and then mixed. The control group without isocitrate was included in the experiment. Group were added 1.25,2.50,3.75,5.00 mmol / L concentration of isocitrate, frozen and resumed after routine sperm analysis. In the meantime, 9 semen samples of healthy males were selected and the fluorescent dyes 5,5 ’, 6,6’ tetrachloro-1 and 1 ’, 3,3’ tetraethylbenzimidazolyl carbonylated iodine (5, 5 ’, 6,6’-tetrachloro-1,1’, 3,3 "-tetraethyl-imidacarbo-cyanine iodide (JC-1) was used to detect the mitochondrial membrane potential of spermatids after single resuscitation. Results: After resuscitation, the percentage of forward motile sperm (PR) in G3 group increased (P <0.01), the level of PR in G2 group increased (P <0.05) and that in G3 group increased (P <0.05) The membrane potential increased compared with the control group (P <0.05). Conclusion: Adding appropriate concentration of isocitrate to the semen cryoprotectant can protect mitochondrial function of sperm after cryopreservation and increase the sperm motility and concentration after resuscitation.