The Effect Of Pentoxifylline On Capacitation And Acrosome Reaction Of Mouse Sperm.

Objectives: To study the effect of Pentoxifylline on capacitation and Acrosome Reaction of mouse sperm.Methods and results: Experiment 1: Mouse sperm was exposed to 0mg/ml, 0.125mg/ml, 0.25mg/ml, 0.5mg/ml, 1mg/ml, 2mg/ml, 4mg/ml PF and 10μg/ml P4 for 30 minutes respectively. Percentage of AR was detected by CBB staining. The effect of different concentration of PF and P4 on AR of mouse sperm was studyed. The dose-effect relationship between PF and AR of mouse sperm was analyzed. It was shown that there was no significant difference(P>0.05) in percentage of AR between the group treated with different concentration of PF and the control group(PF0.0-30),no significant difference(P>0.05) between the group treated with P4 and the control group.Experiment 2: Mouse sperm was exposed to different concentration of PF(as above mentioned) and 10ug/ml P4 by turns for 30 minutes and 10 minutes, respectively in order to induce AR of capacitated spermatozoa. Percentage of AR was detected by CBB staining. The effect of different concentration of PF on AR of mouse sperm induced by P4 was studyed. The dose-effect relationship between PF and capacitation of mouse sperm was analyzed. It was shown that there was no significant difference(P>0.05) in percentage of AR between the group treated with 0.125mg/ml PF and the control group(PF0.0-30;P4-10). However, there was significant difference(P<0.05) in percentage of AR between the group treated with 0.25mg/ml PF and the control group, great significant difference(P<0.01) in percentage of AR between the group treated with 1mg/ml and upwards PF and the control group.Experiment 3: Mouse sperm was incubated by capacitated culture medium for 60 minutes. After that, Mouse sperm was exposed to different concentration of PF, 10μg/ml P4, PF and P4 together, respectively, for 10 minutes. Percentage of AR was detected by CBB staining. The effect of different concentration of PF on AR of capacitated sperm was studyed. The dose-effect relationship between PF and AR of capacitated sperm was analyzed. It was also studyed that whether PF and P4 are involved in synergistic effect on AR of capacitated sperm. It was shown that there was no significant difference(P>0.05) in percentage of AR between the group treated with 0.5mg/ml and downwards PF and the control group(PF0.0-10). However, there was significant difference(P<0.05) in percentage of AR between the group treated with 1mg/ml and upwards PF and the control group, great significant difference(P<0.01) in percentage of AR between the group treated with P4, PF and P4 together and the control group. There was no significant difference(P>0.05) in percentage of AR between the group treated with P4 and the one treated with PF and P4 together.Experiment 4:Mouse sperm was exposed to the lowest concentration of PF that was able to promote the AR of capacitated sperm. Percentage of AR of mouse sperm was detected by CBB staining at Ominute, 20 minute, 40 minute, 60 minute, 80 minute, 100 minute, respectively. The time-effect relationship between PF and AR of mouse sperm was analyzed. It was shown that there was no significant difference(P>0.05) in percentage of AR between the group treated with PF and the control group(M16) at Ominute, 20 minute, 40 minute, respectively. However, the percentage of AR in the group treated with PF was significantly higher than the control group after 60 minutes. Conclusions: 1. PF can promote capacitation of mouse sperm, induced an early onset of sperm capacitation, the lowest effective concentration is 0.25mg/ml.2. 0.5mg/ml and downwards PF could not promote AR of mouse capacitated sperm whereas 1mg/ml and upwards PF could promote it.3. PF and P4 did not have synergistic effect on AR of capacitated spermatozoa.4. Compared to control group, 1mg/ml PF could promote AR of mouse sperm after 60 minutes.