The Initial Exploration Of Endogenous Ouabain In The Pathogenesis Of Asthenospermia

Objective To study the effects of Ouabain and it's receptors: NKA-α1 polyclonal antibody and NKA-α4 polyclonal antibody on human sperm movement.Methods 60 normal semen samples were selected according to the World Health Organization standards. After optimized by swim-up technique, 30 samples were co-incubated with Ouabain of different concentration(1×10-2 M~1×10-7 M), the changes of the sperm movement were detected by CASA, the movement parameters observed contained: sperm motility, sperm activity ratio, curve velocity, whiplash frequency and so on; Another 30 samples were co-incubated with NKA-α1 antibody and NKA-α4 antibody respectively, the movement parameters observed were same as above.Results 1. The sperm motility and sperm activity ratio interfered by Ouabain presented time-effect relationship and they were decreased gradually with the time. The sperm motility was decreased by 62.7%~68.4%(p<0.05)and sperm activity ratio was decreased by 81.5%~85.7%(p<0.01)after co-incubated with high-dose(1×10-2 M~1×10-5 M)Ouabain for 4h; But the changes among the four dose-groups had no statistical significance(p>0.05);Compared with the control group, the sperm motility and sperm activity ratio had no statistical significance (decreased by 27.2±3.7% and 24.5±2.9% respectively )after co-incubated with low-dose(1×10-6 M and 1×10-7 M)Ouabain for 4h(p>0.05);③The rest of movement parameters had no statistical significance. 2.①The sperm activity ratio was decreased by 28.1±2.8% and 2.9±3.4% respectively after co-incubated with Anti-α1 and Anti-α4(p<0.01); But the changes between the two groups had no statistical significance(p>0.05);②The sperm motility was decreased by 38.7±3.8% and 53.2±4.4% respectively after co-incubated with Anti-α1 and Anti-α4(p<0.01); Compared with Anti-α1, the effect of Anti-α4 on sperm motility was dramatically obvious(p<0.01).③The rest of movement parameters had no statistical significance.Conclusion Ouabain and it's receptors: NKA-α1 polyclonal antibody and NKA-α4 polyclonal antibody can degrade the motor function of human sperm in vitro; They mainly decrease sperm activity ratio and sperm motility; Compared with Anti-α1, the effect of Anti-α4 is more obvious, which suggest that Anti-α4 perhaps play the principal role in sperm movement.Objective To detect the EO level in seminal plasma of normal fertile, light and severe asthenospermia males, and to explore the dependablity between EO and asthenospermia and it's course of disease.Methods 60 normal semen samples were selected according to the World Health Organization standards. The other semen of 60 light asthenospermia and 60 severe asthenospermia were selected at the same time. The competitive ELISA examination were performed to detect the EO level in seminal plasma using the anti-ouabain polyclonal antibody made by our laboratory.Results The EO level in seminal plasma of light and severe asthenospermia males were 25.27±1.71μg/L and 26.52±1.82μg/L respectively; They were dramatically higher than normal fertile males 19.32±1.45μg/L(p<0.01); Compared with the light the asthenospermia, the EO level in seminal plasma of severe asthenospermia was slightly higher but had no statistical significance(p>0.05).Conclusion The EO level in seminal plasma of asthenospermia patients is dramatically higher and present certain positive correlation with the course of disease; Combining with the results of Experiment One, it's indicated that EO perhaps participate in the regulation of sperm movement and EO/NKA may be new pathway of athenospermia pathological regulation.Objective To examine the expression and expression feature of EO in the testes of normal fertile and athenospermia SD rats, and to explore the interrelationship between EO and asthenospermia.Methods Select 10 normal fertile and 10 asthenospermia SD rats; Make paraffin sections of testes; The immunohistochemical (IHC) examination was performed using the anti-ouabain polyclonal antibody made by our laboratory.Results The expression of EO displayed in the kytoplasm and cell membrane of Leydig cells, matured spermatogenic cells (round spermatoblast and long spermatoblast)of the normal fertile and athenospermia testes of SD rats, but the expression level of athenospermia was markedly increased (p<0.01).Conclusion EO expresses in the testes of normal fertile and asthenospermia SD rats, and the changes of expression intensity indicate that EO perhaps participate in the pathological regulation.of athenospermia.