Chan P J, Corselli J U, Patton W C, Jacobson J D, King A
Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, California 92350, USA.
J Assist Reprod Genet. 1998 Jan;15(1):32-8. doi: 10.1023/a:1022526305186.
The objectives of this study were (1) to determine the sperm hyperactivation and related kinematic parameters at 40 degrees C after using four sperm wash procedures and (2) to correlate the heat-induced hyperactivation data with cases of clinical pregnancies from either artificial insemination or standard in vitro fertilization (IVF).
Semen samples (n = 51) were collected by ejaculation, and semen analyses were carried out to determine the pretreatment data. Sperm kinematic measurements were performed using the Hamilton Thorn HTM-C computer-aided sperm analyzer. Hyperactivation was determined using the sort module on the HTM-C. Membrane integrity was assessed using the hypoosmotic sperm swelling procedure. Sperm morphology and acrosomal status were also determined using the Spermac stain. Each semen specimen was divided and processed through either the swim-up wash, the 1-h test-yolk buffer (TYB) wash, the 1 mg/ml pentoxifylline stimulant procedure, or the two-layer 90:47% gradient colloidal solution procedure. The washed sperm were incubated at 25 or at 40 degrees C for 4 hr. After incubation, kinematic parameters were assessed for the posttreatment data. Semen specimens were obtained on different occasions for artificial insemination or standard IVF. Data from intracytoplasmic sperm injection cases were not included to avoid confounding factors. Live births and/or pregnancies with fetal heart-beat examined by ultrasound were considered clinical pregnancies.
Heat-induced hyperactive motility was significantly higher in sperm of the male partner of pregnant (n = 7) patients compared with nonpregnant (n = 44) patients (mean +/- SE, 10.0 +/- 3.3 versus 5.5 +/- 0.8%) after TYB processing followed by 4 hr of incubation at 40 degrees C. This was also observed after colloid (Percoll) processing (11.6 +/- 4.6 versus 5.8 +/- 0.8%). There were no differences in hyperactivation after 4 hr at 23 degrees C between pregnant and nonpregnant cases. Parameters such as count, volume, motility, viability, and acrosomal status were not different for the groups. However, the percentage of sperm with normal morphology (WHO classification) was twice as high in the pregnant group versus the nonpregnant group.
Heat-induced hyperactivation was associated with fertile sperm and was predictive of pregnancy obtained after artificial insemination or IVF. The association was evident only after TYB or Percoll sperm processing. The study could not confirm the finding of significant decreases in motility after heat treatment of sperm derived from infertile males. The mechanism for heat-induced hyperactivation did not involve membrane integrity or the sperm acrosome, although an involvement of heat shock proteins was postulated. Interestingly, there were no pregnancies when sperm did not exhibit heat-induced hyperactivation.
本研究的目的是：（1）在使用四种精子洗涤程序后，测定40摄氏度下精子的超激活及相关运动学参数；（2）将热诱导超激活数据与人工授精或标准体外受精（IVF）后的临床妊娠病例相关联。
通过射精收集精液样本（n = 51），并进行精液分析以确定预处理数据。使用汉密尔顿·桑恩HTM-C计算机辅助精子分析仪进行精子运动学测量。使用HTM-C上的分类模块确定超激活。使用低渗精子肿胀程序评估膜完整性。还使用Spermac染色确定精子形态和顶体状态。每个精液标本被分开并通过上浮洗涤、1小时试验卵黄缓冲液（TYB）洗涤、1mg/ml己酮可可碱刺激程序或两层90:47%梯度胶体溶液程序进行处理。洗涤后的精子在25或40摄氏度下孵育4小时。孵育后，评估处理后数据的运动学参数。在不同时间获取用于人工授精或标准IVF的精液标本。不包括来自卵胞浆内单精子注射病例的数据以避免混杂因素。通过超声检查有活产和/或有胎心搏动的妊娠被视为临床妊娠。
在经TYB处理并在40摄氏度下孵育4小时后，与未妊娠（n = 44）患者相比，妊娠（n = 7）患者男性伴侣的精子中热诱导的超激活运动显著更高（平均值±标准误，10.0±3.3对5.5±0.8%）。在胶体（Percoll）处理后也观察到这种情况（11.6±4.6对5.8±0.8%）。在23摄氏度下孵育4小时后，妊娠和未妊娠病例之间的超激活没有差异各组的计数、体积、活力、存活率和顶体状态等参数没有差异。然而，妊娠组中具有正常形态（WHO分类）的精子百分比是未妊娠组的两倍。
热诱导的超激活与有生育能力的精子相关，并且可预测人工授精或IVF后获得的妊娠。这种关联仅在TYB或Percoll精子处理后明显。该研究未能证实不育男性精子热处理后活力显著下降的发现。热诱导超激活的机制不涉及膜完整性或精子顶体，尽管推测热休克蛋白参与其中。有趣的是，当精子未表现出热诱导的超激活时没有妊娠发生。
