烟酰胺腺嘌呤二核苷酸对精子DNA碎片化指数的影响

发布时间：2019-03-30 22:10

【摘要】：目的研究烟酰胺腺嘌呤二核苷酸(NAD)对精子DNA碎片化的作用,并探讨此过程中的分子生物学机制。方法在差异浓度NAD(0、1、10、100、1 000μmol/L)孵育下,观察最适宜的精子NAD孵育浓度;采用Bio Vsion公司生产的NAD/NADH定量试剂盒测定精子NAD水平;采用流式细胞仪观察精子DNA碎片率和染色质成熟度;采用实时定量PCR观察精子DNA单链修复酶PARP1和双链修复酶BRCA1的转录水平。结果 10μmol/L NAD孵育即能显著增加精子NAD含量。在精子内NAD水平显著升高的同时,精子DNA碎片率明显降低,然而精子染色质成熟度不受精子NAD水平的影响。精子内NAD水平能显著影响精子内DNA单链修复酶PARP1和双链修复酶BRCA1的转录水平。结论这些结果初步提示,精子内NAD水平可能通过调节DNA修复系统,参与精子DNA损伤修复。
[Abstract]:Aim to study the effect of nicotinamide adenine dinucleotide (NAD) on the fragmentation of sperm DNA and to explore the molecular biological mechanism in this process. Methods the optimal concentration of sperm NAD was observed at different concentrations of NAD (0,1,10,100,1000 渭 mol / L), and the NAD level of spermatozoa was determined by NAD/NADH quantitative kit produced by Bio Vsion Company. The DNA fragmentation rate and chromatin maturity were measured by flow cytometry, and the transcription levels of DNA single strand repair enzyme PARP1 and double strand repair enzyme BRCA1 were measured by real-time quantitative PCR. Results 10 渭 mol / L NAD significantly increased the content of NAD in spermatozoa. While the level of NAD in spermatozoa increased significantly, the percentage of sperm DNA fragments decreased significantly. However, the sperm chromatin maturity was not affected by the level of NAD in spermatozoa. The level of NAD in spermatozoa significantly affected the transcription level of DNA single strand repair enzyme PARP1 and double strand repair enzyme BRCA1 in spermatozoa. Conclusion these results suggest that the level of NAD in spermatozoa may be involved in the repair of sperm DNA damage by regulating the DNA repair system.
【作者单位】：中国医科大学附属盛京医院妇产科;
【分类号】：R321.1

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