S型氯代甘油醇对大鼠精子运动和超活化抑制作用研究

Inhibitory effect of(S)-chlorohydrin on rat sperm motility and hyperactivation

目的研究S型氯代甘油醇(SACH)对大鼠精子运动和超活化的影响和可能机制。方法 20只成熟雄性SD大鼠随机分为4组,分别给予0、2.5、5.0和10mg/kg BW SACH,连续灌胃染毒52天。取附睾尾精子,获能条件下培养5h,以计算机辅助精子分析系统检测精子运动和超活化运动,同时检测精子特异的3-磷酸甘油醛脱氢酶(GAPDS)活性、三磷酸腺苷(ATP)和环磷酸腺苷(cAMP)水平,观察己酮可可碱(PTF)对SACH的拮抗作用。结果 SACH染毒组大鼠精子曲线运动速率(VCL)、平均路径速率(VAP)、直线运动速率(VSL)和精子头侧摆幅度(ALH)与对照组相比受到显著抑制(P<0.01),运动直线性(LIN)显著增强(P<0.01)。SACH使VCL≥400μm/s或LIN≤20%的比例显著减少(P<0.01和P<0.05),表明精子超活化受抑制。SACH显著抑制GAPDS活性,ATP和cAMP水平也随染毒剂量增加呈下降趋势。PTF可改善精子cAMP水平,部分缓解精子运动和超活化的抑制程度。结论 SACH抑制大鼠精子GAPDS活性导致ATP和cAMP水平下降,可能是影响精子运动和超活化的重要机制。

Abstract：Objective To investigate the effects of （S）-α-chlomhydrin （SACH） on rat epididymal sperm motility and hyperactivation and explore the mechanisms of the effects. Methods 20 adult male Sprague-Dawley rats were divided randomly into 4 groups and dosed orally with 0, 2. 5, 5. 0 and 10mg/kg BW SACH respectively for 52 days. After the cauda epididymal sperm were incubated under a eapaeitating condition for 5h, sperm motility and hyperactivation parameters were obtained by computer-assisted sperm analysis （CASA）, and sperm-specific glyceraldehyde 3-phosphate dehydrogenase （GAPDS） activity, adenosine triphosphate （ATP） and cyclic adenosine monophosphate （cAMP） were assayed. The protecting effect of pentoxifylline （PTF） against SACH was aslo tested. Results The spermfrom the SACH-treated rats treated showed significant decreases in curvilinear velocity （VCL）, average path Velocity （VAP）, straight line velocity （VSL） and amplitude of lateral head movement （ALH） （P 〈 0. 01 of all）, and an increase in linearity （LIN） （ P 〈 0. 01 ）. The SACH-treated rats had much less sperm population with VCL≥400μm/s or LIN ≤20% than that of the control （P 〈 0. 05 and P 〈 0. 01, respectively ）, indicating that SACH diminished hyperactivation of rat sperm. GAPDS activities were inhibited by SACH, and decreasing trends of ATP and cAMP levels were observed. PTF rescued the cAMP level which was depressed by SACH, and alleviated in part the inhibition of sperm motility and hyperactivation. Conclusion SACH impaired the motility and hyperactivation of rat sperm, which might result from the inhibition of GAPDS by SACH and subsequent defects of ATP and cAMP.