男性不妊精子の酸化的DNA障害とその病理学的意義

1997年，我们调查了不育男性患者精子中是否存在高水平的氧化性DNA损伤，并研究了抗氧化剂治疗对这种损伤水平的影响。比较19例不育男性和17例正常男性精子DNA氧化损伤水平。此外，14名不育男性给予抗氧化剂2个月。采用高效液相色谱电化学检测法测定精子中氧化损伤的一种形式8-羟基-2 '-脱氧鸟苷的水平。精子DNA中的8-羟基-2 '-脱氧鸟苷水平在配偶不育患者中显著高于对照组（1.5*0.2对1.0*0.1/10^5脱氧鸟苷），并且与精液中的精子浓度相关。抗氧化剂处理对精子浓度产生了显著的积极影响，精子8-羟基-2 '-脱氧鸟苷水平显著降低（从1.5*0.2降至1.1*0.14/10^5天 ...更多信息 1998年，我们研究了氧自由基在热应激诱导的睾丸生殖细胞凋亡中的作用。将从未成熟大鼠分离的睾丸细胞在高温或非高温下培养，通过琼脂糖凝胶电泳后DNA片段的出现和凋亡细胞的流式细胞术定量来确定这些细胞中凋亡的发生。在32.5 ℃下，< 1%的细胞在整个培养期间显示出凋亡迹象，而在热应激下，在43 ℃下暴露1小时后，随后在32.5 ℃下培养23小时，凋亡细胞的比例增加到14%。与热应激的作用类似，外源性氧自由基也诱导凋亡。与此相反，过氧化氢酶处理显着衰减热应激诱导的细胞凋亡。此外，睾丸细胞热应激与细胞内过氧化物水平升高有关，我们的研究表明精子DNA氧化损伤水平与男性不育之间存在相关性，并表明自由基氧参与热应激诱导的睾丸生殖细胞凋亡。少

In 1997, we investigated whether a high level of oxidative DNA damage in spermatozoa occurs in infertile male patients, and examined the influence of antioxidant treatments on the levels of this damage. The levels of oxidative DNA damage in spermatozoa of infertile male (n= 19) and control patients (n= 17) were compared. In addition, 14 infertile males were given antioxidants for 2 months. The levels of 8-hydroxy-2'-deoxyguanosine, a form of oxidative damage, in the spermatozoa were determined using high performance liquid chromatography with electrochemical detection. The levels of 8-hydroxy-2'-deoxyguanosine in the sperm DNA were significantlyhigher in mate infertile patients than control (1.5*0.2 versus 1.0*0.1/10^5 deoxyguanosine), and were correlated with sperm concentrations in ejaculates. Antioxidant treatment resulted in significant positive effects on sperm concentrations, with a significant reduction in sperm 8-hydroxy-2'-deoxyguanosine levels (from 1.5*0.2 to 1.1*0.14/10^5 d … More eoxyguanosine).In 1998, we investigated the role of radical oxygen species in the testicular germ cell apoptosis induced by heat stress. Testicular cells isolated from immature rats were cultured with or without elevated temperature, and occurrence of apoptosis in these cells was defined by the appearance of DNA fragmentation Following agarose gel electrophoresis, and by the flow cytometric quantification of apoptotic cells. At 32.5ﾟC, < 1% of cells showed signs of apoptosis throughout the culture period, whereas, under heat stress, the proportion of apoptotic cells increased to 14 % after I h exposure at 43ﾟC followed by 23 h culture at 32.5 ﾟC.Similar to the effect of heat stress, exogenously supplied oxygen free radicals also induced apoptosis. In contrast, treatment with catalase significantly attenuated heat stress-induced apoptosis. Furthermore, heat stress of testicular cells was associated with an increased intracellular peroxide level.Our studies demonstrate an association between the level of oxidative DNA damage in spermatozoa and male infertility, and indicate the involvement of radical oxygen species during testicular germ cell apoptosis induced by heat stress. Less