Grunewald S, Paasch U, Wuendrich K, Glander H-J
Division of Andrology, Department of Dermatology, University of Leipzig, Leipzig, Germany.
Arch Androl. 2005 Nov-Dec;51(6):449-60. doi: 10.1080/014850190947813.
The programmed cell death (PCD, the apoptosis) is associated with the activation of the "death enzymes" caspases and very likely plays a role in the decrease of sperm vitality during cryopreservation. The activation of pan-caspases was examined in fresh and cryostored sperm by fluorescence microscopy applying a cell permeable and noncytotoxic carboxy-fluorescein labelled caspase inhibitor. Cryopreservation increased significantly the percentage of spermatozoa with activated pan-caspases (aCP) from 21 +/- 2.6 to 47 +/- 5.8 % (mean +/- SEM). Quantitative Western blot analyses confirmed the activation of caspase-1, -8, and -9 in detail on protein level after cryopreservation, whereas, the caspases of sperm of infertility patients showed a higher activation status than donors' sperm. This effect raised with increasing glycerol concentration from 7-14 %. The higher level in activation of caspases in cryostored spermatozoa of infertility patients may indicate that these cells have a lower cryotolerance and a higher susceptibility to caspase activation than does the sperm of donors.
程序性细胞死亡（PCD，即细胞凋亡）与“死亡酶”半胱天冬酶的激活相关，并且很可能在冷冻保存过程中精子活力降低方面发挥作用。通过使用一种可穿透细胞且无细胞毒性的羧基荧光素标记的半胱天冬酶抑制剂，采用荧光显微镜检查新鲜和冷冻保存精子中泛半胱天冬酶的激活情况。冷冻保存显著增加了激活泛半胱天冬酶（aCP）的精子百分比，从21±2.6%增至47±5.8%（平均值±标准误）。定量蛋白质印迹分析详细证实了冷冻保存后蛋白质水平上半胱天冬酶-1、-8和-9的激活，然而，不育患者精子中的半胱天冬酶显示出比供体精子更高的激活状态。随着甘油浓度从7%增加到14%，这种效应增强。不育患者冷冻保存精子中半胱天冬酶的较高激活水平可能表明，与供体精子相比，这些细胞具有较低的冷冻耐受性和对半胱天冬酶激活的更高敏感性。
