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。
Objective To investigate the effects of nicotinamide adenine dinucleotide （NAD） on sperm DNA fragmentation.Methods Sperms were incubated with 0,1,10,100 or 1 000 μmol/L NAD for screening of the optimum concentration of NAD intervention.NAD levels were determined by using NAD/NADH Quantification Kits according to the manufacturer＇ s instructions.DNA fragmentation index and high DNA stainability were evaluated by flow cytometry.Single and double stranded DNA repair enzyme were analyzed by real time-PCR.Results 10 μmol/L NAD incubation can effectively increase the NAD content in sperms.Sperm DNA fragmentation was significantly decreased,along with the increased sperm NAD levels,but high DNA stainability was not affected.Sperm single and double stranded DNA repair enzyme can be affected by sperm NAD levels.Conclusion Taken together,these results indicate that sperm NAD levels may be involved in sperm DNA damage repair through the regulation of the DNA repair system.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2014年第11期999-1002,共4页
Journal of China Medical University
