Department of Genetics, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
J Assist Reprod Genet. 2012 Jul;29(7):625-30. doi: 10.1007/s10815-012-9755-5. Epub 2012 Apr 11.
The aim of this study was to investigate whether treatment of sperm from infertile patients would gluthatione could reduce sperm premature chromosome condensation (PCC). To reach the goals of this study, the frequency of sperm PCC formation in sperm of normal and sub-fertile men with/without glutathione treatment were compared and analyzed.
Hamster oocytes were retrieved after super ovulation by PMSG and HCG injection. Following treatment with hyaluronidase, zonae was removed by trypsin digestion. Sperm were classified into 3 groups according to morphology, movement and counts, treated with glutathione(10 μg/ml) and then processed by swim up method. After capacitation, zona-free oocytes were incubated with sperm then transferred to fresh media containing colcemid. Cells were fixed and slides prepared using Tarkowskie's standard air drying technique and after staining in 5% Giemsa, oocytes were analyzed at high magnification.
Sperm penetration rate was higher and the rate of intact sperm head and PCC was lower in GSH treated samples compared to non treated groups. Sperm penetration rate was significantly higher in treated astheno sperm samples compared to non-treated ones (66.4% and 50. 97% respectively) (P < 0.001). We observed a significantly higher PCC frequency in infertile patients (P < 0.001). In addition, there was a significantly lower rate of intact sperm head in treated astheno sperm samples (17.49%) compared to non treated ones (26.79%) (P < 0.001). Finally, a significantly lower rate of PCC in treated astheno sperm samples comparing to non treated ones was seen (51.06% and 72.96% respectively) (P < 0. 001).
Our results show that sperm PCC formation could be one of the major causes of failed fertilization in individuals with sperm abnormalities. Also sperm PCC formation may be involved in the etiology of some cases of idiopathic infertility. Given that the susceptibility of sperm to oxidative stress is significantly greater in idiopathic infertile men, our results show that treatment with glutathione could significantly reduce these stress factors and increase ART outcome.
本研究旨在探讨对不育患者精子进行谷胱甘肽处理是否可以降低精子早熟染色体凝聚（PCC）。为了达到本研究的目的，比较并分析了正常和亚生育男性精子在有无谷胱甘肽处理的情况下 PCC 形成的频率。
通过 PMSG 和 HCG 注射超排卵后收集仓鼠卵母细胞。用透明质酸酶处理后，用胰蛋白酶消化去除透明带。根据形态、运动和计数将精子分为 3 组，用谷胱甘肽（10μg/ml）处理，然后用泳动法处理。获能后，将无透明带的卵母细胞与精子孵育，然后转移到含有秋水仙素的新鲜培养基中。细胞用 Tarkowskie 标准空气干燥技术固定，用 5%吉氏染色后，在高倍显微镜下分析卵母细胞。
与未处理组相比，GSH 处理组的精子穿透率更高，完整精子头和 PCC 的比例更低。与未处理组相比，处理后的弱精样本的精子穿透率显著更高（分别为 66.4%和 50.97%）（P<0.001）。我们观察到不育患者的 PCC 频率显著升高（P<0.001）。此外，处理后的弱精样本中完整精子头的比例明显低于未处理组（分别为 17.49%和 26.79%）（P<0.001）。最后，与未处理组相比，处理后的弱精样本的 PCC 比例明显降低（分别为 51.06%和 72.96%）（P<0.001）。
我们的结果表明，精子 PCC 形成可能是精子异常个体受精失败的主要原因之一。此外，精子 PCC 形成可能与一些特发性不孕病例的病因有关。鉴于特发性不育男性对氧化应激的敏感性明显增加，我们的结果表明，用谷胱甘肽处理可以显著降低这些应激因素，提高 ART 结果。
