顶体酶的研究现状及临床应用价值

在诊疗男性不育症的临床实践中,我们逐渐认识到传统的精液分析并不能完全反映精子的受精能力,而精子顶体酶活性作为一种反映精子受精能力的重要指标,逐渐引起更多人的重视。本文将从精子顶体酶的生理作用、影响因素、在诊断和治疗中的价值等四个方面,对目前的研究进展及发展前景进行综述。     

少弱精症患者精子顶体酶活性分析

目的:研究少弱精症不育男性精子顶体酶活性变化与其生育力的关系。方法:以159例少弱精症和25例正常生育男性为研究对象,应用改良Kennedy法检测精子顶体酶活性,分析不同精子密度、活力、形态与顶体酶活性的关系。结果:正常组的顶体酶活性高于不同精子密度不育各组(>50×106ml、20～50×106ml、<20×106ml)(F=7.504,P<0.001);不同精子密度不育各组之间顶体酶活性比较,>50×106ml组顶体酶活性高于<20×106ml组(P<0.05),其它各亚组两两比较顶体酶活性无统计学差异;少弱精症不育者的精子畸形率与顶体酶活性呈负相关(r=-0.171),a+b级精子活力(r=+0.142)和顶体完整率(r=+0.154)与顶体酶活性呈正相关。结论:顶体酶活性对少弱精症男性生育力评价有重要价值,顶体酶活性可以作为精子形态分析之外重要的预测精子受精能力的指标。     

形态正常精子百分率与顶体酶活性的关系

目的:探讨精子形态与精子顶体酶活性的关系。方法:对737例男性不育患者采用分光光度比色法测定顶体酶活性,采用精子形态检测系统下人工修正方法分析精子形态。结果:精子形态正常组的顶体酶活性明显高于精子形态异常组,两组间比较差异显著(P<0.05);顶体酶活性正常组形态正常精子百分率明显高于顶体酶活性异常组形态正常精子百分率,两组间比较差异亦具有显著性(P<0.001);顶体酶活性与形态正常精子百分率间呈显著正相关关系(r=0.172,P<0.001)。结论:精子形态可影响精子顶体酶活性。     

精子DNA碎片率及顶体酶活性对男性不育的诊断价值

目的：分析精子DNA碎片率、顶体酶活性诊断男性不育的效用。方法：选择2016年6月至2021年12月入院诊断的1727例男性筛查者,其中男性不育患者481例。根据DNA碎片指数(DFI)分组,一组的DFI值≤10%;二组为10%6为27例,>48.2μU/10⁶为416例,对比各项参数差异。结果：不同DFI组间的年龄、精子密度、顶体酶活性、前向运动率、正常精子率以及非前向运动率对比存在差异P<0.05。根据顶体酶活性分组,对比组间的精子密度、前向运动率、DFI、正常精子率与非前向运动率有差异P<0.05。相关系分析可见,DFI与精子密度、前向运动率、正常精子率、顶体酶活性具有负相关性,顶体酶活性与精子密度、前向运动率、正常精子率具有正相关性,与DFI具有负相关性。结论：DFI与顶体酶活性能够反映精子质量,可以通过以上指标诊断男性不育,具有较高的诊断价值。     

135名不育厨师精液质量变化的分析

目的探讨厨师职业对男性精液质量的影响.方法对2495名男性不育者(厨师135名、非厨师2360名)和100名男性生育者,依据EHO提倡的人类精液实验室检验的方法和标准,作全面的精液分析.结果精液质量异常率,不育厨师组为71.1%,比非厨师职业不育组的62.3%和生育组的11.0%有显著增加(P＜0.05和P＜0.01);厨师操厨≥5年组精液质量异常率(82.8%)亦显著高于＜5年组(62.3%,P＜0.01);随操厨年限的延长,精液质量的下降主要表现在精子的形态(P＜0.01)、活动力(P＜0.01)、成活率(P＜0.05)和密度(P＜0.05)等方面.结论厨师的职业环境可能是引起男性精液质量下降、造成男性不育的因素之一.     

冷冻对精子顶体酶活性及精子功能影响的研究

目的:探讨精子冷冻对精子功能和精子顶体酶活力的影响.方法:选取来自正常生育男性志愿者,且精液常规分析各参数正常的精液为标本,检测精子冷冻前后精液常规各参数、精子功能和精子顶体酶活性.结果:冷冻后精子存活率54.96±6.00％,精子宫颈黏液穿透能力1.70±0.47,精子顶体酶活性15.31±8.87mU/百万精子,与冷冻前精子存活率65.74±6.62％,精子宫颈黏液穿透能力2.65±0.49,精子顶体酶活性54.32±36.38mU/百万精子比较显著下降(P＜0.05).结论:精子冷冻导致精子膜功能和精子穿透功能受损,引起精子顶体酶活性显著下降.     

用顶体反应释放的顶体酶活性评价顶体反应功能的研究

目的:研究用顶体反应释放的顶体酶活性评价人类精子顶体反应功能。方法:10例健康生育男子精液,采用Percoll密度梯度分离法优选精子,用FITC-PSA荧光染色法分析顶体反应(acrosome reaction,AR),顶体酶活性采用分光光度比色法测定。结果:A23187和孕酮诱导的精子AR释放的顶体酶活性和顶体反应率均明显增加(P<0.05)。随A23187浓度增加,AR释放的顶体酶活性和顶体反应率均明显增加(P<0.01)。A23187诱导AR释放的顶体酶活性与相应顶体反应率呈明显正相关关系(r=0.950,P<0.001)。随孕酮浓度增加,AR释放的顶体酶活性和顶体反应率均明显增加(P<0.05)。孕酮诱导的AR释放的顶体酶活性与相应顶体反应率亦呈明显正相关关系(r=0.921,P<0.001)。结论:孕酮可诱导人精子顶体反应,用顶体反应释放的顶体酶活性可评价人精子顶体反应功能。     

Y染色体平衡易位与精液质量的相关性研究

目的 分析Y染色体平衡易位与男性精液质量的相关性。方法 回顾性分析2019年1月—2021年12月北京大学第三医院生殖医学中心就诊的男性病例资料,筛选出14例染色体核型分析为Y染色体平衡易位病例,以及同期185例染色体核型结果正常(46, XY)的男性作对照。通过比较两组间精液常规质量,包括精子的数量、活率以及形态学检测,分析Y染色体与常染色体间平衡易位与男性精液质量的相关性。结果 除精液量外,Y染色体平衡易位患者的精液密度、精子总数、精子活率、前向运动精子百分率以及正常形态精子百分率均显著低于对照男性组。14例患者中有7例为无精子症,7例为严重少弱畸形精子症。q12(14例)、q11.23(4例)是Y染色体平衡易位断裂点高发区域。结论 Y染色体平衡易位对于精子数量、活力及形态具有显著影响,严重时可导致少弱畸形精子症或无精症,影响患者的生育能力。     

甲氧滴滴涕对大鼠精子顶体酶活性的影响

目的观察甲氧滴滴涕(methoxychlor,MXC)对雄性大鼠精子顶体酶活性的影响,以探讨其生殖毒性的可能机理。方法在精子悬液中加入不同浓度的MXC,以改良的Kennedy法分别测定不同浓度MXC作用下的精子顶体酶活性。结果与对照组比较,MXC作用后的各组精子顶体酶活性明显降低,且降低幅度与MXC的作用浓度有关。结论MXC对大鼠精子顶体酶活性具有明显的抑制作用。     

顶体酶抑制剂EGB的急性毒性,阴道粘膜刺激性和阴道乳酸杆菌抑菌实验研究

本文对一种新合成的精子顶体酶抑制剂EGB的急性毒性,阴道粘膜刺激性和对阴道乳酸杆菌的影响进行了实验研究。研究结果表明:EGB 小鼠腹腔注射给药LD_(50)为228.9±20.9mg/kg体重,口服灌胃给药LD_(50)>5g/kg体重。对局部阴道粘膜的刺激性在可接受范围之内。当给药浓度高达100mg/ml时对人阴道乳酸杆菌的生长繁殖仍无抑制作用。因此,作者认为EGB作为局部阴道外用避孕药是安全,可靠的。     

Evaluation of the Sperm Motility Analyzer System (SMAS) for the Assessment of Sperm Quality in Infertile Men

Eighty-one fresh semen samples were analyzed to compare the sperm parameters obtained using the new Sperm Motility Analysis System (SMAS; version 1.0, Kaga Electronics, Tokyo, Japan) with the CellSoft^TM Series 3000 (CRYO Resources, New York, USA) computer-assisted semen analysis (CASA) system and conventional manual semen analysis, based on WHO guidelines.. Significant correlations of sperm concentration (p<0.0001) and sperm motility (p<0.0001) were observed between SMAS and manual semen analysis estimates. There were also significant correlations of sperm concentration (p=0.0003) and sperm motility (p<0.0001) between SMAS and CellSoft estimates. Significant correlations for motility-related parameters were demonstrated in sperm velocity (p<0.0001), and linearity (linear velocity (VSL) divided by curvilinear velocity (VCL)×100) (p<0.0001), amplitude of lateral head displacement (ALH) (p<0.0001), and beat/cross frequency (BCF) (p=0.0127), between SMAS and CellSoft estimates. In this study, we showed the usefulness of the new SMAS, which has high reliability in estimating sperm concentration, sperm motility, velocity and linearity compared with CellSoft. SMAS can be a promising alternative, providing cost-effective semen analysis with the utility of the CASA system.     

男性不育患者精液分析和精子形态学分析

目的：回顾性分析助孕门诊男性不育患者精液分析与精子形态学检测，为防止临床对女方过度治疗提供依据。方法：选择2007年10月1日～2008年2月29日在本院助孕门诊就诊的不孕夫妇，男方至少行精液分析2次，在第一次精液分析即进行精子形态学分析。精液分析采用清华同方精液分析系统，精子形态学分析采用改良巴氏法染色，应用Microsoft Excel进行分析。结果：进行精液分析共1206人次，进行精子形态学分析共517人次。精子密度0-180^★10^9／L，平均55．3±32．2^★10^9／L，75．1％的病例精液密度为20-100^★10^9／L，仅13．3％为少精症，无精症占1．5％；a级精子为14．0％±9．9％，b级精子为10．6％±7．8％，a级＋b级〉50％者仅占0．6％，超过半数的为30％-50％（51．1％），精子形态学分析，仅有20．5％病例是正常的（〉14％）。结论：除无精症显然与不育相关外，精液密度可能不是评估男性不育的一个很好的指标，精子活动力低下或精子畸形率过高更能预示不孕因素，精液分析和精子形态学分析可以作为助孕门诊男性不育的首要筛查手段，防止对女方的过度治疗。     

精索静脉曲张合并不育症的中西医结合临床研究进展

对于精索静脉曲张合并不育症治疗手段纷繁复杂。目前,采用西医手术结合中医中药综合治疗此病的治疗方案逐渐被认可。本文综述了精索静脉曲张合并不育症的西医治疗方案、中医治疗方案及现代检验学在此病中的应用,并对此治疗方案的发展方向进行展望。     

Current Assessment of Sperm DNA Integrity

Conventional semen analysis is rapidly losing its place as the gold standard of diagnosis and the cornerstone of treating the infertile male in modern times. Recent technology allows scientists to analyze sperm fertilizing potential and subsequent embryonic growth by studying factors that have previously escaped traditional parameters. It has become increasingly evident that nuclear DNA arrangement is essential to the fertilizing potential of sperm. A vast array of tests are now available to examine the genetic makeup of individual spermatozoa, ranging the entire gamut from simple bench top assays performed routinely to complex flow cytometric assays requiring highly-skilled technologists. Future research to compare these new tests to those more commonly in use, correlating them with reproductive outcome promises to fill the current void in the field of male infertility, paring innovative diagnostic (and prognostic) technological standards to the already existing sophisticated assortment of successful treatment modalities.     

精子顶体发育相关蛋白的研究进展

受精是新生命诞生的第一步,在此过程中,精子将所携带的单倍体遗传物质与卵子的单倍体遗传物质融合成双倍体合子。哺乳动物的受精过程涉及到一系列复杂而精细的活动,如精子的超活化与获能、顶体反应以及精卵结合等。精子顶体是一种特殊的膜细胞器,具有帽状结构并覆盖在精子的细胞核前部,在受精过程中发挥着关键作用。顶体的形成主要包括囊泡形成、囊泡运输、囊泡融合和顶体与细胞核结合等过程,其独特的蛋白运输机制需要内质网、高尔基体等细胞器以及某些特殊结构之间的相互配合。顶体形成是精子细胞分化为精子的关键步骤之一,受多基因精细调控,其发育缺陷与包括圆头精子症在内的多种男性不育症有关。综述近年精子顶体发育过程中主要生物学事件的发生顺序中相关蛋白的研究,并介绍了自噬相关蛋白对顶体发育过程作用的新近研究,以期为男性不育症的诊疗提供新思路。     

精液质量评估与男性生育力关系

目的不育夫妇中,50%与男性因素相关,随着不育症的发病率增高,对男性生育力的正确评估显得尤为重要。《世界卫生组织人类精液检查与处理实验室手册》的更新不断推动着男科实验室的发展,也提出了问题与挑战。正确认识精液质量评估的各种参数与男性生育力之间的关系对临床诊断以及治疗方案的选择至关重要。     

Sperm epigenetics in the study of male fertility,offspring health,and potential clinical applications

The mammalian sperm contains a highly unique and specialized epigenetic landscape that offers a great degree of interesting research opportunities. One key discriminating feature of the mature sperm epigenome is that it, in theory, represents both remnant marks used throughout spermatogenesis to generate sperm cells competent to perform their function, but also marks that appear to be useful beyond fertilization. Key questions must be asked about the utility of these marks and the multiple purposes that may be served. It is this unique epigenetic landscape that has driven some labs to begin to study the links between aberrant sperm epigenetic patterns and various forms of infertility, from idiopathic to alterations in sperm motility, morphology, and viability and fertilization capacity. Because of the unique nature of the sperm epigenome and the patterns found in mature sperm that appear to reflect perturbations in spermatogenesis that may ultimately have effect on pregnancy outcomes, some researchers believe that these marks may provide predictive insight that can be exploited. Indeed, there is emerging data suggesting that the predictive power of DNA methylation and RNA signatures in sperm likely exceeds that which can be found with traditional assessments of male infertility. This review will focus on the utilization of the sperm epigenome as a potential diagnostic tool in the context of male infertility, as well as the potential difficulties associated with such an approach.     

High Throughput,Parallel Imaging and Biomarker Quantification of Human Spermatozoa by ImageStream Flow Cytometry

Spermatid specific thioredoxin-3 protein (SPTRX-3) accumulates in the superfluous cytoplasm of defective human spermatozoa. Novel ImageStream technology combining flow cytometry with cell imaging was used for parallel quantification and visualization of SPTRX-3 protein in defective spermatozoa of five men from infertile couples. The majority of the SPTRX-3 containing cells were overwhelmingly spermatozoa with a variety of morphological defects, detectable in the ImageStream recorded images. Quantitative parameters of relative SPTRX-3 induced fluorescence measured by ImageStream correlated closely with conventional flow cytometric measurements of the same sample set and reflected the results of clinical semen evaluation. Image Stream quantification of SPTRX-3 combines and surpasses the informative value of both conventional flow cytometry and light microscopic semen evaluation. The observed patterns of the retention of SPTRX-3 in the sperm samples from infertility patients support the view that SPTRX3 is a biomarker of male infertility.