| Background and Purposes: Male infertility accounts for about 50% of the total number of infertility in the world,and affects about 10% of all men.Recent studies have shown that PDE5 inhibitors can increase sperm motility.In this study we planed to explore the effect of different concentrations of vardenafil pretreatment on weak sperm in vitro,including sperm function,sperm morphology and sperm cell membrane integrity,the rate of sperm DNA fragments,and the effect of vardenafil pretreatment on the post-thaw sperm activity.This study will provide evidence for the application of PDE5 inhibitors in assisted reproductive technologies,such as cryopreservation of sperm,diagnosis of male infertility,in vitro treatment and artificial insemination.Materials and Methods: Part Ⅰ: In vitro effects of vardenafil pretreatment on the viability of idiopathic asthenospermia Semen samples of 30 male patients were collected,according to the exclusion and inclusion criteria.Each semen sample was divided into 5 tubes,one was regarded as the control group,the other 4 groups were added with the prepared vardenafil solution.The final concentration of vardenafil was 0.4μg /m L,4μg /m L,40μg /m L,and 400μg /m L,respectively.The test tubes were gently oscillated and incubated at 37℃ water bath kettle for 0min,15 min,30min,60 min and 90 min.Then the sperm concentration and sperm motility were detected by the WLJY-9000 sperm quality detection system according to the WHO manual of human semen examination and treatment laboratory(the fifth edition).Sperm DNA fragment index(DFI)was detected by acridine orange staining.Sperm morphology was strictly interpreted by papanicolaou staining according to WHO manual of human semen examination and treatment laboratory(fifth edition),and the percentage of normal sperm was evaluated.Sperm viability was detected by eosin staining.According to above results,the optimal concentration and treatment time of vardenafil were determined,and the contents of c AMP and c GMP in sperm were detected by immunospectrophotometry to analyze the changes of each index.Part Ⅱ: Effect of vardenafil pretreatment on the activity of post-thaw sperm According to the exclusion criteria and inclusion criteria,the semen samples of 30 healthy donors from Shanghai human sperm bank were collected.The final concentration of vardenafil was 0μg /m L,0.4μg /m L,4μg /m L,40μg /m L,400μg /m L,and incubated at room temperature for 5 min.The ratio of semen to protectant was 2:1.After balancing at room temperature for 5 min,it was put into a 1.0 m L frozen tube and directly into CRYOPLUS 1 one-step fumigation apparatus.After freezing,it was put into liquid nitrogen for preservation.After 24 h,semen specimen was removed immediately at 37 ℃ water bath for 5 min.WLJY-9000 sperm quality detection system was used to detect sperm motility and percentage of forward moving sperm according to WHO manual of human semen examination and treatment laboratory(fifth edition).The improved pap staining method was used to strictly interpret the sperm morphology and evaluate the percentage of normal sperm according to the WHO manual of human semen examination and treatment laboratory(fifth edition).Results:Part Ⅰ: In vitro effects of vardenafil pretreatment on the viability of idiopathic asthenospermia After treatment with vardenafil,sperm motility was significantly increased at a concentration of 4μg /m L for 30min(P<0.01),normal sperm percentage showed no statistical difference compared with the control group(P>0.05),sperm activity was increased compared with the control group(P<0.05),and sperm DNA fragmentation index(DFI)showed no statistical difference compared with the control group(P>0.05).And the content of c AMP and c GMP in sperm was significantly increased(P<0.01)Part Ⅱ Effect of vardenafil pretreatment on the activity of post-thaw sperm After thawing,the progressive sperm rates were(41.47±9.80)%,(42.57±9.60)%,(47.77±8.55)%,(37.27±8.47)%,and(26.37±6.99)% in the groups treated with 0,0.4,4.0,40.0,and 400.0 μg/m L of vardenafl,respectively.Compared to the control group(0 μg/m L of vardenafl),4.0 μg/m L of vardenafl could signifcantly improve the post-thaw sperm motility(P=0.034).Conclusions: 1.Vardenafil can significantly improve the sperm motility of patients with asthenospermia in vitro,with the optimal concentration of 4μg /m L.This study provides a new drug treatment basis for clinical asthenospermia.Whether vardenafil can be used as a treatment for infertility with low sperm motility,and as an agonist for sperm motility for artificial insemination or in vitro fertilization,which remains to be studied.2.Vardenafl pretreatment can significantly improve the activity of the postthaw sperm. |
PDF Full Text Request