Armstrong J S, Rajasekaran M, Hellstrom W J, Sikka S C
Department of Urology, Tulane University Medical Center, New Orleans, Louisiana, USA.
J Androl. 1998 Jul-Aug;19(4):412-9.
Human serum albumin (HSA) is being considered as an alternate media for sperm enrichment in assisted reproductive technology (ART) because of recent concern with the use of Percoll. In this study, we compared HSA and Percoll for 1) sperm recovery, 2) reactive oxygen species scavenging potential, and 3) effects on total oxidative stress to spermatozoa. The spermatozoa-enriched fractions obtained from Percoll (80%:40%) and HSA (12%) were monitored for sperm motility, viability, hypoosmotic swelling test (HOST), and adenosine triphosphate (ATP) levels. The effect of superoxide anions (O2.-) on donor human spermatozoa was observed in the presence of either HSA or Percoll media. A combination of luminol and the Cypridina luciferin analog 2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyraz in-3-one hydrochloride was used as a highly sensitive chemiluminescence probe in our hypoxanthine and xanthine oxidase-based assay for O2.-. Sperm membrane total oxidative stress was determined by measuring levels of the prostanoid 8-iso-Prostaglandin F2alpha (8-iso-PGF2alpha). Significant differences in sperm parameters between the Percoll-enriched spermatozoa (motility 60%+/-4%, viability 56%+/-6%, and HOST 73%+/-7%) and those enriched with HSA (motility 84%+/-5%, viability 85%+/-4%, and HOST 84%+/-3%; P < 0.01) were observed. Adenosine triphosphate levels were significantly higher, by almost 50%, in samples processed with HSA than with Percoll (P=0.03). The dismutation rate of O2.- in HSA (slope -6.8) was significantly lower than in Percoll (slope -87.0; P < 0.01). Sperm motility and ATP levels decreased at a slower rate after treatment with O2.- in the presence of HSA when compared to Percoll; moreover, spermatozoa in HSA regained partial motility after 2 hours, whereas spermatozoa in Percoll were immobilized. No significant differences in 8-iso-PGF2alpha levels in spermatozoa enriched by either HSA or Percoll were observed. We conclude that the HSA sperm enrichment procedure improves the recovery of higher quality spermatozoa compared to Percoll and, because of its antioxidant properties, may be useful in processing high leukospermia semen samples for ART purposes.
由于最近对使用 Percoll 的担忧，人血清白蛋白（HSA）正被视为辅助生殖技术（ART）中用于精子富集的替代介质。在本研究中，我们比较了 HSA 和 Percoll 在以下方面的差异：1）精子回收率；2）活性氧清除潜力；3）对精子总氧化应激的影响。对从 Percoll（80%:40%）和 HSA（12%）获得的精子富集部分进行精子活力、存活率、低渗肿胀试验（HOST）和三磷酸腺苷（ATP）水平的监测。在 HSA 或 Percoll 介质存在的情况下，观察超氧阴离子（O2.-）对供体人类精子的影响。在我们基于次黄嘌呤和黄嘌呤氧化酶的 O2.- 测定中，使用鲁米诺和海萤荧光素类似物 2 - 甲基 - 6 -（4 - 甲氧基苯基）- 3,7 - 二氢咪唑并（1,2 - α）吡嗪 - 3 - 酮盐酸盐的组合作为高灵敏度化学发光探针。通过测量前列腺素 8 - 异前列腺素 F2α（8 - iso - PGF2α）的水平来确定精子膜的总氧化应激。观察到 Percoll 富集的精子（活力 60%±4%，存活率 56%±6%，HOST 73%±7%）与 HSA 富集的精子（活力 84%±5%，存活率 85%±4%，HOST 84%±3%；P < 0.01）在精子参数上存在显著差异。用 HSA 处理的样本中的三磷酸腺苷水平比用 Percoll 处理的样本显著高近 50%（P = 0.03）。HSA 中 O2.- 的歧化率（斜率 - 6.8）显著低于 Percoll（斜率 - 87.0；P < 0.01）。与 Percoll 相比，在 HSA 存在的情况下用 O2.- 处理后，精子活力和 ATP 水平下降的速度较慢；此外，HSA 中的精子在 2 小时后恢复了部分活力，而 Percoll 中的精子则被固定。在 HSA 或 Percoll 富集的精子中，未观察到 8 - iso - PGF2α 水平的显著差异。我们得出结论，与 Percoll 相比，HSA 精子富集程序提高了高质量精子的回收率，并且由于其抗氧化特性，可能有助于处理用于 ART 目的的高白细胞精液样本。
