Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa
Analysis of DNA fragmentation, plasma membrane translocation of phosphatidylserine and oxidative stress in human spermatozoa
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DOI：
10.1093/humrep/15.6.1338
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发表时间：
2000-06-01
影响因子：
6.1
通讯作者：
Oehninger, S
中科院分区：
文献类型：
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作者：
Barroso, G;Morshedi, M;Oehninger, S
这项横断面观察性研究的目的是：(1)检测DNA。分析高活力和低活力纯化精子群体中磷脂酰丝氨酸的损伤和质膜移位；以及(Ii)分析它们与内源性产生活性氧物种的关系。不育男性精液经梯度离心法检测，主要观察指标为：精子运动参数(计算机分析仪测定)、活性氧产生(化学发光法测定)、DNA损伤(用末端脱氧核苷酸转移酶介导的dUDP缺口末端标记法和单链DNA单抗标记法检测)和膜磷脂酰丝氨酸移位(膜联蛋白V染色检测)。所有患者在低精子活力组和高精子活力组均可观察到DNA断裂和磷脂酰丝氨酸膜移位。低精子活力组的DNA损伤细胞比例和产生活性氧的比例显著高于高精子活力组(P<；0.005)。结论：(1)膜联蛋白V(Annexin V)阳性染色证实，不育男性精子存在膜磷脂异丝氨酸易位；(2)不育男性精液DNA损伤(DNA断裂和单链DNA的存在)存在于精子活力低和活力高的部位，(3)DNA损伤与氧化应激有关。
The objectives of this cross-sectional observational study were: (i) to detect DNA. damage and plasma membrane translocation of phosphatidylserine in purified sperm populations of high and low motility; and (ii) to analyse their relationship with the endogenous generation of reactive oxygen species. Ejaculates from infertile men were examined following gradient centrifugation, The main outcome measures were: sperm motion parameters (assessed with a computer analyser), generation of reactive oxygen species (measured by chemiluminescence), DNA damage (detected by terminal deoxvnucleotidyl transferase-mediated dUDP nick-end labelling and monoclonal antibody labelling of single-stranded DNA) and translocation of membrane phosphatidylserine (examined with annexin V staining). DNA fragmentation and membrane translocation of phosphatidylserine were observed in the fractions with low and high sperm motility in all patients. The fractions with low sperm motility had significantly higher proportion of cells with DNA damage and production of reactive oxygen species than the fractions with high sperm motility (P < 0.005). DNA fragmentation was significantly and positively correlated with the generation of reactive oxygen species (r = 0.42; P = 0.02), Zn conclusion: (i) spermatozoa from infertile men display translocation of membrane phosphatidyIserine as diagnosed by annexin V positive staining; (ii) DNA damage (fragmentation and presence of single-stranded DNA) can be detected in ejaculated spermatozoa from infertile men in fractions with low and high sperm motility, and (iii) there is a relationship between DNA, damage and oxidative stress.