Duru N K, Morshedi M, Schuffner A, Oehninger S
The Jones Institute for Reproductive Medicine, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk, Virginia 23507, USA.
Fertil Steril. 2000 Oct;74(4):715-20. doi: 10.1016/s0015-0282(00)01494-1.
To assess the effects of progesterone and acetyl-L-carnitine used before semen cryopreservation-thawing on sperm motility parameters and plasma membrane integrity.
Prospective cohort study.
Academic tertiary center.
PATIENT(S): Subfertile men undergoing semen evaluation.
INTERVENTION(S): Before cryopreservation, spermatozoa were incubated with water-soluble progesterone (1 and 10 microM), acetyl-L-carnitine (2.5, 5, 10, and 20 mM), or both (progesterone, 1 microM; and acetyl-L-carnitine, 5 mM).
MAIN OUTCOME MEASURE(S): Postthaw change of motility parameters (computer-assisted measurements) and vitality-membrane integrity (examined with eosin-Y staining and annexin V-Cy3 binding assay).
RESULT(S): There were no statistically significant differences between control samples and samples treated with progesterone and/or acetyl-L-carnitine for cryosurvival rate, motility parameters, or membrane integrity. The percentages of postthaw cells identified as live showed significantly different results with use of the eosin-Y staining and annexin V binding assay.
CONCLUSION(S): Neither progesterone nor acetyl-L-carnitine seemed to prevent cryodamage assessed by motility changes or membrane integrity in human spermatozoa of subfertile men. Annexin V binding, a reflection of membrane translocation of phosphatidylserine, provided more distinct information about postfreezing membrane integrity changes than eosin-Y staining.
评估精液冷冻保存-解冻前使用孕酮和乙酰左旋肉碱对精子活力参数和质膜完整性的影响。
前瞻性队列研究。
学术三级中心。
接受精液评估的不育男性。
在冷冻保存前,精子与水溶性孕酮(1和10微摩尔)、乙酰左旋肉碱(2.5、5、10和20毫摩尔)或两者(孕酮1微摩尔;乙酰左旋肉碱5毫摩尔)一起孵育。
解冻后活力参数(计算机辅助测量)的变化以及活力-膜完整性(用伊红-Y染色和膜联蛋白V-Cy3结合试验检测)。
对照样本与用孕酮和/或乙酰左旋肉碱处理的样本在冷冻存活率、活力参数或膜完整性方面无统计学显著差异。用伊红-Y染色和膜联蛋白V结合试验检测解冻后活细胞的百分比显示出显著不同的结果。
对于不育男性的人类精子,孕酮和乙酰左旋肉碱似乎均不能通过活力变化或膜完整性评估来预防冷冻损伤。膜联蛋白V结合试验,即磷脂酰丝氨酸膜易位的反映,比伊红-Y染色能提供关于冷冻后膜完整性变化更清晰的信息。