氰戊菊酯对小鼠精子体外获能的影响及机制探讨

Effect of Fenvalerate on in vitro Sperm Capacitation in Mice and its Mechanisms

[目的 ]观察氰戊菊酯 (fenvalerate ,Fen)对小鼠精子体外获能的影响并探讨其作用机制。 [方法 ] 2 0 0 μl小鼠精子细胞悬液加入不同终浓度Fen染毒后 ,CO2 孵箱 3 7℃培育 2h ,以金霉素 (chlortetracycline ,CTC)荧光染色后涂片观察小鼠精子体外获能培养后B型精子的发生率。[结果 ]以终浓度为 0、0 .62 5、1.2 5、2 .5、5 .0、10 .0、2 0 .0 μmol·L-1的Fen处理小鼠精子 ,其B型精子发生率分别为 ( 5 5 .0 5± 0 .43 ) %、( 5 2 .0 2± 1.2 9) %、( 4 7.13± 0 .87) %、( 4 0 .0 8± 0 .3 2 ) %、( 3 5 .79± 0 .88) %、( 2 8.66± 0 .96) %、( 2 7.89± 1.5 6) % ,半数有效剂量 (ED50 )为 5 μmol·L-1,其作用呈剂量 反应关系。同时Fen以时间依赖方式抑制B型精子发生率。进一步观察到 2 .5、5 .0、10 .0、2 0 .0 μmol·L-1的Fen组小鼠精子 ,与对照组相比 ,其精子细胞膜脂质流动性下降 [以 1,6 二苯已三烯 ( 1,6 diphenyl 1,3 ,5 hexatriene ,DPH )为荧光探针 ] ,作用呈剂量 反应关系。 [结论 ]氰戊菊酯抑制小鼠精子体外获能 ,其作用机制主要是氰戊菊酯降低了精子细胞膜脂质流动性。

To observe the effect of fenvalerate,a pyrethroid insecticide,on in vitro sperm capacitation in mice,and to study the mechanisms of its action. Mouse in vitro sperm capacitation was assessed by the B pattern of staining by chlortetracycline(CTC),then the fluidity parameters were monitored to reveal the dynamic changes in sperm membrane due to fenvalerate interaction by measuring the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene(DPH) incorporated into the membrane. Sperm capacitation was inhibited by fenvalerate in a concentration-dependent and time-dependent manner. The ED 50 is 5μmol·L -1 . Fenvalerate was also found to increase the DPH fluorescence polarization value in a concentration-dependent manner,implicating that it makes the membrane less fluid. [Conclusion] Fenvalerate inhibits sperm capacitation by decreasing membrane fluidity in mice sperm.