Gomez E, Irvine D S, Aitken R J
MRC Reproductive Biology Unit, Edinburgh, UK.
Int J Androl. 1998 Apr;21(2):81-94. doi: 10.1046/j.1365-2605.1998.00106.x.
A spectrophotometric assay for the measurement of malondialdehyde and 4 hydroxyalkenals (MA + 4HA) has been evaluated for the detection of sperm pathologies involving oxidative stress. In order to make sensitive measurements of MA + 4HA on human spermatozoa, the stimulation of a lipid peroxidation cascade with a ferrous ion promoter was found to be necessary. The optimal configuration for the promoter was defined (0.64 mM FeSO4 + 20 mM ascorbate for 2 h in Ca2+ and Mg2 free Hanks' balanced salt solution) and the assay used in a series of studies to elucidate the functional significance of MA + 4HA determinations. Such measurements were found to give highly significant correlations (p < 0.001) with the loss of motility induced by oxidative stress created either with a xanthine oxidase, free radical generating system or by prolonged incubation under aerobic conditions. Experiments involving the stimulation and suppression of lipid peroxide release from human sperm suspensions, in concert with a bioassay for cytotoxicity, confirmed the strength and causative nature of these associations. Measurements of lipid peroxidation potential in highly purified, leucocyte-free sperm suspensions revealed the presence of inverse correlations with the motility of the spermatozoa, their viability, their competence for sperm-oocyte fusion and, most significantly, the quality of sperm movement in the original semen samples. Similar negative correlations were observed between sperm function and phorbol ester-stimulated reactive oxygen species generation but, unlike the MA + 4HA determinations, these relationships were obfuscated by the presence of leucocytes. We conclude that the measurement of MA + 4HA in human spermatozoa provides important information on the underlying quality of spermatogenesis and should be of value in the clinical diagnosis of infertility involving oxidative stress and the selection of patients for antioxidant therapy.
一种用于测量丙二醛和4-羟基烯醛（MA + 4HA）的分光光度测定法已被评估用于检测涉及氧化应激的精子病理。为了对人类精子中的MA + 4HA进行灵敏测量，发现用亚铁离子启动剂刺激脂质过氧化级联反应是必要的。确定了启动剂的最佳配置（在不含Ca2+和Mg2+的汉克斯平衡盐溶液中，0.64 mM硫酸亚铁 + 20 mM抗坏血酸，作用2小时），并在一系列研究中使用该测定法来阐明MA + 4HA测定的功能意义。发现这种测量与由黄嘌呤氧化酶、自由基产生系统或在有氧条件下长时间孵育产生的氧化应激诱导的运动能力丧失具有高度显著的相关性（p < 0.001）。涉及刺激和抑制人类精子悬液中脂质过氧化物释放的实验，与细胞毒性生物测定法相结合，证实了这些关联的强度和因果性质。在高度纯化、无白细胞的精子悬液中测量脂质过氧化潜能，发现与精子的运动能力、活力、精卵融合能力以及最显著的是原始精液样本中精子运动质量呈负相关。在精子功能与佛波酯刺激的活性氧生成之间也观察到类似的负相关，但与MA + 4HA测定不同的是，这些关系因白细胞的存在而变得模糊。我们得出结论，测量人类精子中的MA + 4HA可提供有关精子发生潜在质量的重要信息，在涉及氧化应激的不孕症临床诊断以及选择抗氧化治疗患者方面应具有价值。
