Kawaguchi Takashi, Kawachi Mamoru, Morikawa Makoto, Kazuta Hiroki, Shibata Kenji, Ishida Mayumi, Kitagawa Naoko, Matsuo Asaki, Kadota Toshihito
Department of Toxicology and Safety Assessment, Kawanishi Pharma Research Institute, Nippon Boehringer Ingelheim Co., Ltd., 3-10-1 Yato, Kawanishi, Hyogo 666-0193, Japan.
J Toxicol Sci. 2004 Aug;29(3):217-31. doi: 10.2131/jts.29.217.
This study was undertaken to detect key parameters of rat sperm motion in relation to male fertility by comparing the differences in sperm motion induced by treatment with alpha-chlorohydrin (ACH), known to produce spermatotoxicity, and nitrobenzene (NTB), known to produce testicular toxicity. Male rats received ACH (5 or 20 mg/kg/day) or NTB (60 mg/kg/day) for either 3 days or 18 days. Epididymal sperm was assessed for motility using a Hamilton-Thorne Sperm Analyzer (HTM-IVOS). Numerical data for statistical analysis and graphical renditions of sperm motion using parameters in radar charts and reconstructed sperm tracks were analyzed to evaluate sperm motion. Males were allowed to copulate with untreated females and cesarean sections were conducted in order to examine the effects of drug administration on male fertility. Linearity of sperm track (linearity (LIN) and/or straightness (STR)) decreased and/or beat cross frequency (BCF) increased only in ACH groups (5 or 20 mg/kg/day), although the percentage of motile sperm, sperm velocities (average path velocity (VAP), curvilinear (VCL), and straight line velocity (VSL)) and amplitude of lateral head displacement (ALH) decreased on Day 18 in both ACH and NTB (60 mg/kg/day) groups. Furthermore, from the individual reconstructed sperm tracks, it was clear that ACH-treated spermatozoa were characterized by abnormal motion ("jerking") with low vigor (low velocities) and little or no forward progression. Finally, only ACH treatment led to a reduction in pregnancy rate or infertility. Therefore, our results suggest that linearity (especially VSL, STR and LIN) in sperm motion is a key parameter for assessing a chemical's potential to induce male infertility.
本研究旨在通过比较已知会产生精子毒性的α-氯醇（ACH）和已知会产生睾丸毒性的硝基苯（NTB）处理所诱导的精子运动差异，来检测与雄性生育力相关的大鼠精子运动关键参数。雄性大鼠接受ACH（5或20毫克/千克/天）或NTB（60毫克/千克/天）处理，为期3天或18天。使用汉密尔顿-桑恩精子分析仪（HTM-IVOS）评估附睾精子的活力。分析用于统计分析的数值数据以及使用雷达图参数和重建精子轨迹的精子运动图形表示，以评估精子运动。让雄性大鼠与未处理的雌性大鼠交配，并进行剖腹产以检查药物给药对雄性生育力的影响。仅在ACH组（5或20毫克/千克/天）中，精子轨迹的线性度（线性度（LIN）和/或直线度（STR））降低和/或摆动交叉频率（BCF）增加，尽管在第18天，ACH组和NTB（60毫克/千克/天）组中活动精子的百分比、精子速度（平均路径速度（VAP）、曲线速度（VCL）和直线速度（VSL））以及头部侧向位移幅度（ALH）均降低。此外，从个体重建的精子轨迹可以明显看出，ACH处理的精子具有异常运动（“抽搐”）的特征，活力低（速度低），几乎没有或没有向前推进。最后，只有ACH处理导致妊娠率降低或不育。因此，我们的结果表明，精子运动的线性度（特别是VSL、STR和LIN）是评估化学物质诱导雄性不育潜力的关键参数。
