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目的比较葡萄糖6磷酸异构酶(GPI)和烯醇化酶1(ENO1)在人类精子中不同耐冻能力组之间表达的差异,寻找预测人类精子冻融后活力可能的标志物。方法收集42例合格供精志愿者的新鲜精液样品,依据冻融后精子活力的不同,将符合条件的样品分为耐冻组和不耐冻组,应用蛋白质印迹(Western blot)技术比较两组样品GPI和ENO1蛋白在表达量上的差异,利用免疫荧光技术验证Western blot结果并观察两种蛋白在精子中的分布。结果在两组样品之间,GPI和ENO1都表现出明显的差异。GPI和ENO1在耐冻组的表达量均高于不耐冻组(分别为P<0.01,P<0.05),而蛋白在精子中的分布未见明显组间差异。结论 GPI和ENO1可以作为标志物在冻存前预测人类精子冷冻复苏后的活力。
Objective To compare the differences in expression of glucose 6-phosphate isomerase (GPI) and enolase 1 (ENO1) among different cryoprotectant groups in human sperm, and to find possible markers for predicting viability of human sperm after freezing-thawing. Methods Fresh semen samples from 42 qualified volunteers were collected. According to the difference of sperm motility after freezing and thawing, eligible samples were divided into anti-freezing group and non-freezing group. Western blotting was used to compare the two groups The differences of the expression of GPI and ENO1 in the samples were analyzed by immunofluorescence to confirm the Western blot results and observe the distribution of the two proteins in the sperm. Results Between the two groups of samples, GPI and ENO1 showed significant differences. The expression levels of GPI and ENO1 were higher in cryopreserved group than in non-freezing group (P <0.01, P <0.05, respectively), but there was no significant difference in the distribution of protein in sperm. Conclusion GPI and ENO1 can be used as markers to predict the vitality of human sperm after cryopreservation before cryopreservation.