摘要目的 研究超低温冷冻对人精子糖被的损伤.方法 利用凝集素芯片比较精子冷冻前、后的凝集素结合谱.结果 通过比较冷冻保护剂和冷冻方法对精子复苏率的影响,确定了CryoSpermTM以及一步熏蒸法为最佳精予冷冻试剂和方法;通过凝集素芯片比较精子冷冻前、后的凝集素结合谱观察到在91种凝集素中,有33种发生了显著变化,其中,冷冻后显著下调的有9种,显著上调的有24种,并且显著下调的MAA及显著上调的PSA、ABA和AIA经流式验证,结果与芯片一致.结论 精子糖被经过冷冻之后发生了显著变化,对精子起保护作用的唾液酸大量丢失,同时也暴露糖链内部的糖基,因此精子糖被的冷冻损伤可能是由于低温冷冻引起精子生育力低下的原因之一.

abstractsObjective To study the effects ofcryopreservation on human sperm glycocalyx.Methods The lectin binding profilings of sperm after frozen-thawed were compared by lectin microarray.Results CryoSpermTM and direct fumigation were confirmed to be the optimized cryoprotectant and method by comparing the sperm recovery rate.In 91 lectins,33 lectins were significantly changed after sperm cryopreservation.Among them,9 lectins were greatly decreased and 24 lectins were mainly increased in frozen-thawed sperm.The binding signals of MAA,PSA,ABA and AIA were verified by FACS,and the results were consistant with that of lectin microarray.Conclusion Sperm glycocalyx occurred significant changes after cryopreservation.The sialic acid,playing an important role on protecting sperm,was seriously lost,which made the inner carbohydrates exposed.So the glycocalyx impairment might be one reason of low sperm fertility due to the cryopreservation.