A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acet...A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen(ROS) level after freezing-thawing process. The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia. The cryopreservation of human spermatozoa treated with acetyl-L-carnitine at different concentrations(group B: 2.5 mmol/L, group C: 7.5 mmol/L, group D: 15 mmol/L) was compared with control(group A: no acetyl-L-carnitine given). For the frozen-thawed spermatozoa, the viability, motility and DNA integrity were measured by comet assay, acrosome integrity by FITC-PNA staining and ROS level was determined in each group. The results showed that there were no significant differences in motility and viability between group A and group B, while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A. As compared with group A, the values for DNA integrity parameters including comet rate(CR), tail DNA percentage(TD), tail length(TL) and Oliver tail moment(OTM) were significantly reduced in group C and group D. Group C and group D also displayed a higher proportion of intact acrosome than group A. No significant difference in ROS level was found between group A and group B, while with the increase in acetyl-L-carnitine concentration, the ROS level in groups C and D was significantly reduced as compared with that in group A. In conclusion, acetyl-L-carnitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.展开更多
[Object]This study aimed to examine the effects of 3,7-dimethyl-1-(5-oxo-hexyl)-xanthine(pentoxifylline,PF)on the motility,mitochondrial activity,acrosome integrity and fertilization rate of spermatozoa in both fr...[Object]This study aimed to examine the effects of 3,7-dimethyl-1-(5-oxo-hexyl)-xanthine(pentoxifylline,PF)on the motility,mitochondrial activity,acrosome integrity and fertilization rate of spermatozoa in both fresh and frozen-thawed bull semen.[Method]Fresh and frozen-thawed bull spermatozoa were exposed to 5 mmol/L PF with untreated samples as controls.[Result]Fresh spermatozoa showed reduced(P〈0.05)motility after 2 h incubation with PF whereas,surprisingly,frozen-thawed samples exhibited increased sperm motility(P〈0.05)after 2 h incubation with PF and they also showed enhanced longevity compared to controls.Mitochondrial activity in both fresh and frozen-thawed bull spermatozoa increased(P〈0.05)during 4 h incubation with PF whereas acrosome integrity remained unchanged in both types of semen.However,treatment with 5 mmol/L PF did not influence the in vitro fertilization efficiency of fresh spermatozoa but improved significantly(P〈0.05)that of frozen-thawed spermatozoa.[Conclusion]These results indicate that PF can improve sperm quality of frozenthawed bull semen,and may improve pregnancy rates in bovine artificial insemination programmes employing frozen semen.展开更多
基金supported by grants from the Natural Science Foundation of Hubei Province(No.2016CFB352)Hubei Province Health and Family Planning Scientific Research Project(No.WJ2017M011)China Scholarship Council(No.201706275124)
文摘A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen(ROS) level after freezing-thawing process. The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia. The cryopreservation of human spermatozoa treated with acetyl-L-carnitine at different concentrations(group B: 2.5 mmol/L, group C: 7.5 mmol/L, group D: 15 mmol/L) was compared with control(group A: no acetyl-L-carnitine given). For the frozen-thawed spermatozoa, the viability, motility and DNA integrity were measured by comet assay, acrosome integrity by FITC-PNA staining and ROS level was determined in each group. The results showed that there were no significant differences in motility and viability between group A and group B, while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A. As compared with group A, the values for DNA integrity parameters including comet rate(CR), tail DNA percentage(TD), tail length(TL) and Oliver tail moment(OTM) were significantly reduced in group C and group D. Group C and group D also displayed a higher proportion of intact acrosome than group A. No significant difference in ROS level was found between group A and group B, while with the increase in acetyl-L-carnitine concentration, the ROS level in groups C and D was significantly reduced as compared with that in group A. In conclusion, acetyl-L-carnitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.
基金Supported by NMKJ Project of Production and Industrial Application of Sexed Semen in Domestic Animals(No.20111701)
文摘[Object]This study aimed to examine the effects of 3,7-dimethyl-1-(5-oxo-hexyl)-xanthine(pentoxifylline,PF)on the motility,mitochondrial activity,acrosome integrity and fertilization rate of spermatozoa in both fresh and frozen-thawed bull semen.[Method]Fresh and frozen-thawed bull spermatozoa were exposed to 5 mmol/L PF with untreated samples as controls.[Result]Fresh spermatozoa showed reduced(P〈0.05)motility after 2 h incubation with PF whereas,surprisingly,frozen-thawed samples exhibited increased sperm motility(P〈0.05)after 2 h incubation with PF and they also showed enhanced longevity compared to controls.Mitochondrial activity in both fresh and frozen-thawed bull spermatozoa increased(P〈0.05)during 4 h incubation with PF whereas acrosome integrity remained unchanged in both types of semen.However,treatment with 5 mmol/L PF did not influence the in vitro fertilization efficiency of fresh spermatozoa but improved significantly(P〈0.05)that of frozen-thawed spermatozoa.[Conclusion]These results indicate that PF can improve sperm quality of frozenthawed bull semen,and may improve pregnancy rates in bovine artificial insemination programmes employing frozen semen.