烟酰胺腺嘌呤二核苷酸(NAD)对人类精子活力、DNA和顶体酶的保护作用  

Protective effects of nicotinamide adenine dinucleotide(NAD)on human sperm motility,DNA and acrosin

作　　者：房芃[1] 胡利文[1] 赵秀红[1] 陈永刚[1] 王爱妍 毕敬磊 韩昊辰 崔艳国[1] Fang Peng;Hu Liwen;Zhao Xiuhong;Chen Yonggang;Wang Aiyan;Bi Jinglei;Han Haochen;Cui Yanguo(Reproductive Center of Zibo Maternal and Child Health Hospital,Zibo 255000,Shandong,China)

机构地区：[1]淄博市妇幼保健院生殖医学中心,山东淄博255000

出　　处：《中国男科学杂志》2023年第1期36-40,共5页Chinese Journal of Andrology

基　　金：淄博市重点研发计划(2019ZC010009)。

摘　　要：目的探讨烟酰胺腺嘌呤二核苷酸(NAD)及其前体物质烟酰胺腺嘌呤单核苷酸(NAM)对人类精子活力、DNA和顶体酶活性的保护作用。方法选取在我院生殖医学中心就诊的男性精液标本50例,检测其活力(PR级)、DNA碎片指数(DFI)和顶体酶活性。按照NAD/NAM的添加时机不同将每份精液均分为冻前干预组和复苏后干预组。将冻前干预组的标本均分为三份:分别加入PBS、NAD和NAM后37℃孵育30 min,检测其精子活力、DFI和顶体酶活性三项指标后加入精子冷冻保护剂冷冻大于24 h后复苏,再次检测该三项指标。复苏后干预组的标本直接加入精子冷冻保护剂冷冻大于24 h后复苏,复苏后的标本均分为三份:分别加入PBS、NAD和NAM后37℃孵育30 min,检测其精子活力、DFI和顶体酶活性三项指标。另选取男性精液标本38例,每例均分为两份,一份冷冻24 h后复苏、另一份冷冻大于两年后复苏;复苏后的标本均分为三份:分别加入PBS、NAD和NAM后37℃孵育30 min,检测其三项指标。结果精液在冷冻前加入NAD/NAM孵育后精子活力增强、顶体酶活性升高(P<0.05),DFI无明显变化(P>0.05);复苏后活力(前向运动精子百分率)明显降低(30.5%±4.5%)vs(63.3%±6.8%),精子顶体酶活性显著降低(78.2±8.5μIU)vs(135.0±8.2μIU)(P<0.05),DFI显著升高(25.5%±7.5%)vs(10.2%±8.3%)(P<0.05),但冻前NAD/NAM孵育组的精子活力显著高于对照组(38.4%±8.4%)/(36.5%±7.4%)vs(30.5%±4.5%)(P<0.05),顶体酶活性显著高于对照组(90.4±6.5μIU)/(92.5±5.8μIU)vs(78.2±8.5μIU)(P<0.05),DFI显著低于对照组(16.5%±6.8%)/(18.8%±8.7%)vs(25.5%±7.5%)(P<0.05)。复苏后干预组的精液经NAD/NAM孵育后精子活力升高(40.2%±6.3%)/(42.2%±8.8%)vs(31.2%±7.5%);顶体酶活性升高(98.5±7.6μIU)/(93.8±8.2μIU)VS(80.3±5.8μIU)(P<0.05),DFI未见明显变化(P>0.05)。冷冻大于两年后复苏组的精子活力低于冷冻24 h复苏组(24.5%±4.7%)vs(33.5%±7.4%);顶体酶活性也出现类�Objective To investigate the protective effects of nicotinamide adenine dinucleotide(NAD)and its precursor nicotinamide adenine mononucleotide(NAM)on human frozen sperm motility,DNA and acrosin activity.Methods Total of 50 male semen samples were collected from reproductive medicine center of our hospital in the study,the sperm motility,DNA fragmentation index(DFI)and acrosin activity were detected.All the semen samples were classified into pre-freezing intervention group and post-resuscitation intervention group according to the different added time of NAD/NAM.The samples of the pre-freezing intervention group were further divided into three subgroups:they were treated with PBS,NAD and NAM,respectively and incubated at 37℃for 30 min.The sperm motility(PR level),DFI and acrosin activity were detected.After frozen with cryoprotectant for more than 24 h,the three indexes in samples were detected again.The samples in the post resuscitation intervention group were directly added with sperm cryoprotectant,frozen for more than 24 hours,and then resuscitated.The recovered samples were divided into three parts:and they were treated with PBS,NAD and NAM and incubated at 37℃for 30 min,and then the sperm motility,DFI and acrosin activity were detected.Additionally,38 male semen samples were collected,each case was divided into two parts.One was resuscitated after 24 hours of freezing,the other was resuscitated after freezing for more than two years.The recovered samples were divided into three parts,and they were treated with PBS,NAD and NAM and incubated at 37℃for 30 min,then the three indexes in the samples were detected.Result After treated with NAD/NAM before freezing,sperm motility and acrosin activity in the samples were increased(P<0.05),but no significant change in DFI was found(P>0.05);After resuscitation,sperm motility and acrosin activity were significantly decreased[(30.5±4.5%)vs(63.3±6.8%)][(78.2±8.5μIU)vs(135.0±8.2μIU)](P<0.05),and DFI was significantly increased[(25.5±7.5%)vs(10.2±8.3%)](P<0.05)

关 键 词：烟酰胺腺嘌呤二核苷酸(NAD) 精液保存 精子能动性 DNA碎片率 顶体蛋白酶 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]