Taylor S L, Weng S L, Fox P, Duran E H, Morshedi M S, Oehninger S, Beebe S J
The Jones Institute for Reproductive Medicine, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk, Virginia, USA.
Mol Hum Reprod. 2004 Nov;10(11):825-34. doi: 10.1093/molehr/gah099. Epub 2004 Oct 1.
In this study we extended earlier work to determine whether sperm respond to somatic cell apoptotic stimuli and whether apoptotic phenotypes are significant indicators of human sperm quality. We evaluated ejaculated sperm from fertile donors and subfertile patients following purification of fractions of high and low motility. In unstimulated conditions, caspase enzymatic activity was higher in motile fractions from subfertile patients than in donors, and was higher in low motility fractions from both groups. Staurosporine, but not a Fas ligand or H2O2, significantly increased caspase activity, but only in high motility fractions. Procaspase-3, -7 and -9 and low levels of active caspase-3, -7 and -9 were identified by immunoblot analysis. Apoptosis-inducing factor (AIF) was present in all samples but poly ADP-ribose polymerase-1 (PARP-1) was not detected. Phosphatidylserine translocation was significantly increased only with H2O2 treatment. In ejaculates of both subfertile and fertile men, we demonstrated the presence and activation of several proteins that are key constituents of apoptosis-related pathways in somatic cells, which may serve as markers for sperm quality.
在本研究中，我们拓展了早期的工作，以确定精子是否对体细胞凋亡刺激作出反应，以及凋亡表型是否是人类精子质量的重要指标。我们对来自生育力正常的供体和生育力低下的患者的射出精子进行了评估，这些精子经过了高活力和低活力部分的纯化。在未受刺激的条件下，生育力低下患者的活动部分精子中的半胱天冬酶酶活性高于供体，且两组低活力部分精子中的该活性更高。星形孢菌素能显著增加半胱天冬酶活性，但仅在高活力部分精子中起作用，而Fas配体或过氧化氢则无此作用。通过免疫印迹分析鉴定出了前半胱天冬酶-3、-7和-9以及低水平的活性半胱天冬酶-3、-7和-9。所有样本中均存在凋亡诱导因子（AIF），但未检测到聚ADP-核糖聚合酶-1（PARP-1）。仅在过氧化氢处理时，磷脂酰丝氨酸易位显著增加。在生育力低下和生育力正常男性的射精样本中，我们证实了几种蛋白质的存在和激活，这些蛋白质是体细胞凋亡相关途径的关键组成部分，可能作为精子质量的标志物。
