ZK 93426对小鼠和人精子获能的影响研究

Effects of ZK 93426 on mouse and human sperm capacitation

目的探究小分子化合物ZK 93426对小鼠和人精子获能的影响及作用机制。方法采用蛋白质印迹技术(Western Blot)检测ZK 93426对小鼠精子和人精子获能相关的酪氨酸磷酸化的影响;运用Western Blot检测ZK 93426在正常以及缺少HCO3^-,BSA,Ca^2+的获能缓冲液(modified Krebs-Ringer bicarbonate medium,HMB)中对获能的作用;选取精子特异钙离子通道CatSper抑制剂NNC 55-0396及PKA通路抑制剂H89进一步探索ZK 93426对于精子获能的机制。结果 ZK 93426对小鼠精子和人精子获能相关蛋白酪氨酸磷酸化均有促进作用,且在缺少Ca^2+的获能缓冲液中具有更明显的促进蛋白酪氨酸磷酸化作用。NNC 55-0396不会抑制ZK 93426对于获能相关的酪氨酸磷酸化的促进作用,而PKA抑制剂H89可抑制其作用。结论 ZK 93426对精子获能相关的蛋白酪氨酸磷酸化的促进作用不依赖CatSper通道,可能通过作用于PKA通路的上游发挥相应的作用。

Objective To explore the effects of small molecule compound ZK 93426 on mouse and human sperm capacitation and its relevant mechanism. Methods The effects of ZK 93426 on tyrosine phosphorylation related to mouse and human sperm capacitation were detected by Western Blot. The effects of ZK 93426 on sperm capacitation in normal modified Krebs-Ringer bicarbonate medium(HMB) or HMB buffer lack of HCO3^-,BSA or Ca^2+respectively were analyzed by Western Blot. CatSper inhibitor NNC 55-0396 and PKA inhibitor H89 were used to explore the possible mechanism of ZK93426 on sperm capacitation. Results ZK 93426 could promote capacitation-related protein tyrosine phosphorylation in both mouse and human sperms, and exhibited a more significant effect in Ca^2+-deficient HMB. NNC 55-0396 had no effect on ZK93426-promoted sperm capacitation, whereas PKA inhibitor H89 could inhibit its effect. Conclusion ZK 93426 promote the sperm capacitation-related protein tyrosine phosphorylation independent of CatSper channel, but it may play a corresponding role by acting upstream of the PKA pathway.