Carver-Ward J A, Jaroudi K A, Einspenner M, Parhar R S, al-Sedairy S T, Sheth K V
Department of Pathology and Laboratory Medicine, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia.
Hum Reprod. 1994 Jan;9(1):71-6. doi: 10.1093/oxfordjournals.humrep.a138323.
The study was designed to investigate the effects of pentoxifylline on the acrosome reaction of human spermatozoa in vitro, and to determine whether the reaction is differently modulated after sperm selection by multiple tube swim-up and Percoll buoyant density centrifugation. The acrosome reaction was induced in vitro by using calcium ionophore (A23187) and was detected by measuring the fluorescence of FITC-conjugated goat anti-mouse immunoglobulin bound to CD46 antibody (which binds to the CD46 receptor site on the inner acrosomal membrane) by flow cytometry. Spermatozoa separated on Percoll displayed significantly lower spontaneous acrosome reactions (P = 0.002) than did those separated by the swim-up technique. Pentoxifylline did not, by itself, induce acrosome reaction, but after induction with ionophore, it significantly increased the reaction (P = 0.003) and this increase was seen to be greater when Percoll separation was used as compared to the swim-up technique (P = 0.0002). We therefore conclude that Percoll selection of motile spermatozoa together with pentoxifylline treatment may be of value in assisted reproductive techniques, as an increased ARIC score arose after both treatments, and that flow cytometry allows a precise and rapid quantification of the acrosome reaction.
本研究旨在探讨己酮可可碱对人精子体外顶体反应的影响，并确定在通过多管上游法和Percoll浮力密度离心法进行精子筛选后，顶体反应是否受到不同调节。体外使用钙离子载体（A23187）诱导顶体反应，并通过流式细胞术测量与结合CD46抗体（其与顶体内膜上的CD46受体位点结合）的异硫氰酸荧光素（FITC）偶联的山羊抗小鼠免疫球蛋白的荧光来检测。在Percoll上分离的精子显示出自发顶体反应显著低于通过上游技术分离的精子（P = 0.002）。己酮可可碱本身不会诱导顶体反应，但在用离子载体诱导后，它会显著增加顶体反应（P = 0.003），并且与上游技术相比，当使用Percoll分离时，这种增加更为明显（P = 0.0002）。因此，我们得出结论，Percoll选择活动精子并结合己酮可可碱治疗在辅助生殖技术中可能具有价值，因为两种治疗后顶体反应指数（ARIC）得分均有所提高，并且流式细胞术能够精确、快速地定量顶体反应。
