抗精子抗体对精子顶体酶活性的影响

本文旨在观察抗精子抗体对精子顶体酶活性的影响。选择男性不育者５０例与正常生育者２０例，采用固相酶染色法测抗精子抗体，以固定明胶薄膜法测精子顶体酶活性。结果发现５０例不育者抗精子抗体阳性率为５２％，不育者精子顶体酶活性明显低于生育者；抗精子抗体阳性者顶体酶活性明显低于阴性者，表明抗精子抗体可降低精子顶体酶活性。     

肿瘤坏死因子对人精子顶体酶和顶体反应的影响及机制探讨

目的 研究肿瘤坏死因子(TNF-α)对正常精子顶体酶活性和顶体反应的影响及其机制。方法采用BAEE/ADH联合法测定顶体酶和三色染色法技术测定顶体反应。结果 TNF-α可显著降低精子顶体酶的活性和顶体反应(P<0.01;P<0.01),并且它可使精子中的Ca2+-ATPase和SOD的活性显著降低(P<0.05;P<0.001);但TNF-α可使精子中NOS活性增强及NO含量增加(P<0.001;P<0.001);对Na+-K+-ATPase活性影响不明显(P>0.05)。结论 TNF-α对精子顶体酶及顶体反应有一定的抑制作用,并且可能通过降低Ca+-ATPase活性,自由基和NO及增加NOS等多种途径来实现的。     

抗精子抗体对精子顶体酶活性的影响

目的；观察抗精子抗体对精子顶体酶活性的影响。方法：选择男性不育者５０例，与正常生育者２０例。采用固相酶染色法测搞精子抗体，固定明胶薄膜法测精子顶体酶活性。结果：５０例不育者抗精子抗体阳性率为５２５，不育者精子顶体酶生明显低于生育者；抗精子抗体阳怀者顶体酶活性低于阴性者。结论：抗精子抗体可降低精子顶体酶活性。     

抗精子抗体对精子顶体酶活性的影响

目的：观察抗精子抗体对精子顶体酶活性的影响。方法：选择男性不育者５０例，与正常生育者２０例。采用固相酶染色法测抗精子抗体，固定明胶薄膜法测精子顶体酶活性。结果：５０例不育者抗精子抗体阳性率为５２％。不育者精子顶体酶活性明显低于生育者；抗精子抗体阳性者顶体酶活性低于阴性者。结论：抗精子抗体可降低精子顶体酶活性。     

抗精子抗体对精子顶体酶活性的影响

本文旨在观察抗精子抗体对精子顶体酶活性的影响。选择男性不育者50例与正常生育者20例，采用固相酶染色法测抗精子抗体，以固定明胶薄膜法测精子顶体酶活性。结果发现50例不育者抗精子抗体阳性率为52％。不育者精子顶体酶活性明显低于生育者；抗精子抗体阳性者须体酶活性明显低于阴性者，表明抗精子抗体可降低精子顶体酶活性。     

切除颌下腺对大鼠睾酮和精子顶体酶活力的影响

本实验对大鼠切除颌下腺３０和４８天时，睾丸和附睾重量、血清睾酮水平及精子顶体酶活力进行了观察。结果显示：（１）切除颌下腺的实验组，睾丸和附睾重量明显低于对照组（Ｐ＜０．０５，Ｐ＜０．０１），睾丸脏器系数也明显降低（Ｐ＜０．０１），但附睾脏器系数无明显降低（Ｐ＞０．０５）；（２）血清睾酮水平３０天时实验组略低于对照组，４８天时为２．４１ｎｍｏｌ／Ｌ，明显低于对照组的４．４９ｎｍｏｌ／Ｌ（Ｐ＜０．０１）；（３）实验组的精子顶体酶活力分别为７．１９和６．００ｍＵ／ｍｌ明显低于对照组（Ｐ＜０．０１）分别降低了５５．１％和６１．１％。结果表明表皮生长因子（ＥＧＦ）参与了调节睾酮的分泌，并能影响精子顶体酶活力以及睾丸和附睾的重量。     

γ-氨基丁酸对精子顶体酶活性的影响

目的 :研究γ 氨基丁酸 (GABA)对生育男性精子及抗精子抗体 (AsAb)阳性病人的精子顶体酶活性的影响。　方法 :用BAEE ADH联合法同时测定GABA对精子顶体酶活性的影响。　结果 :GABA可明显提高正常及AsAb阳性精子顶体酶的活性 (P <0 .0 1) ;并使精子的Na+ K+ ATPase、Ca2 + ATPase、超氧化物歧化酶 (SOD)的活性增加(P <0 .0 1,P <0 .0 5 ,P <0 .0 0 1)。　结论 :GABA对精子顶体酶活性有显著影响     

人精子透明质酸酶活性测定的临床意义

目的 :测定人精子透明质酸酶 (HYD)活性并分析其与精液常规参数之间的相关性。　方法 :用改良Singer法测定 14 6例男性精子HYD活性 ,常规检测精子密度、活动率和正常形态百分率。　结果 :精子HYD活性与密度、活动率及与正常形态百分率存在显著相关性 (r值分别为 0 .65、0 .63和 0 .72 ,P均 <0 .0 1)。不育男性精子密度在 4 0× 10 6 /ml以上组的HYD活性明显高于精子密度少于 2 0× 10 6 /ml以下组 (P <0 .0 1)。不育男性精子活动率 >60 %组的HYD活性明显高于 <3 0 %组 (P <0 .0 1)。　结论 :精子HYD活性测定是评价精子功能的有效指标之一     

IL-2对精子顶体酶及一氧化氮合酶活性的影响

目的　探讨白细胞介素 2对正常人及白细胞精子症精子功能的影响。方法　精子顶体酶的活性检测采用BAEE/ADH联合法。结果　白细胞精子症及正常人精子顶体酶的活性分别为 (2 7.6 8± 10 .0 6mU/ml,n =18;4 3.6 3± 7.6 1mU/ml,n =18) (P <0 .0 1) ;白细胞精子症及正常人精子一氧化氮合酶分别为 (2 .74± 0 .39U/mg pr,n =18;2 .74± 0 .39U/mgpr,n =18) (P <0 .0 1)。结论　白细胞介素 2对精子顶体酶和一氧化氮合酶均有明显抑制作用 ,提示这可能是导致精子受精能力下降的原因。     

人精子顶体酶活性及其影响因素分析

为研究人精子顶体酶活性与男性不育的关系，分别测定门诊３５６例不育男性精液（实验组）和１８例正常生育者精液（对照组）的顶体酶活性，同时测定实验组男性的精子活力、精子活率、畸形率、镜下白细胞数、精浆抗精子抗体并进行解脲支原体培养，作相关性分析。结果：实验组和对照组顶体酶活性为１０７±９２和２９７±１４３μＩＵ／１０６精子，组间比较差异有显著性（Ｐ＜０００１），顶体酶活性与镜下白细胞数、畸形率明显负相关，与精子活率、精子活力正相关，顶体酶活性与解脲支原体感染有关，但精浆抗精子抗体的存在不影响顶体酶活性。提示顶体酶活性与精子质量有关，是影响生育的重要因素     

NO通过人精子顶体酶对顶体反应的影响

目的探讨NO通过精子顶体酶(acrosin)对项体反应(acrosome reaction,AR)的影响。方法采用BAEE/ADH法测定精子顶体酶的活性,以及通过FITC-PSA法检测精子顶体反应。结果NO供体SNP可诱导人精子表达顶体酶活性,同时促进入精子顶体反应；顶体酶抑制剂TLCK能抑制顶体酶活性,并可抑制SNP诱导的人精子顶体反应。结论NO可能通过调节人精子顶体酶的活性而诱导精子的顶体反应。     

Reactive oxygen species influence the acrosome reaction but not acrosin activity in human spermatozoa

It is now widely accepted that the higher levels of reactive oxygen species (ROS) produced by damaged or deficient spermatozoa are associated with a loss of motility and a decreased capacity for sperm-oocyte fusion. Furthermore, earlier studies show, under physiological conditions, that some ROS may be involved in capacitation and hyperactivation of human spermatozoa. We measured ROS levels, acrosome reaction (AR) and acrosin activity (AA) in semen samples from suspected subfertile men to reveal the influence of ROS on AR and AA of human spermatozoa. Semen samples were obtained from 60 patients. Samples with > or = 1 x 10(6) leukocytes/mL were excluded from the study. ROS production was determined using a chemiluminescence technique. AR was determined using a triple stain technique. The percentage of acrosome-reacted spermatozoa after low temperature induction of the AR (test value), and the inducibility of AR (= the difference between the test value and the control), were calculated. The AA was analysed by determining the proteolytic potential of spermatozoa on gelatin plates. The mean halo diameter and percentage of halo formation in each sample were measured as AA parameters. Scatter plots of ROS levels and AR parameters showed that the percentage of acrosome reacted spermatozoa and AR inducibility were better in samples with low rather than high ROS levels. On the other hand, there were no apparent similarities between ROS and the AA parameters. Therefore, the percentage of acrosome-reacted spermatozoa and AR inducibility were significantly higher in the low than in the high ROS group (p = 0.028, p = 0.0001, respectively). In addition, there was no significant difference in AA parameters between groups. These findings suggest that lower ROS in semen may have a role in AR but excessive ROS may exert a negative influence on AR, while ROS in semen has no relationship to AA.     

The fate of acrosomal staining during the acrosome reaction of human spermatozoa as revealed by a monoclonal antibody and PNA-lectin

A monoclonal anti-human sperm antibody, raised against an acrosomal antigen and indicated to recognize in boar sperm the serine protease, acrosin, stained in human spermatozoa a 50 Kd antigen and several others in the region 24-34 Kd by immunoblotting. The 50 Kd band and the region of 30-34 Kd showed proteolytic activity by zymographic enzyme detection. The fate of the antigen was studied in the acrosome reaction induced by the calcium ionophore A23187. In control incubations 69.5 +/- 14.2% (mean +/- SD) of the spermatozoa had intact acrosomal staining according to indirect immunofluorescence using this antibody whereas in acrosome-reacted samples only 21.0 +/- 2.0% of the sperm were stained. Another marker for the acrosome, peanut agglutinin-lectin (PNA), was used to detect the acrosome with similar results. Acrosome reactions were verified by electron microscopy. The present results indicate that the corresponding antigen, evidently acrosin, and PNA-positive material are liberated during the acrosome reaction which suggests that they are not bound to the inner acrosomal membrane but are components of the acrosomal matrix.     

Effect of leptin on motility, capacitation and acrosome reaction of human spermatozoa

Leptin is a polypeptide hormone with important roles in reproduction. It has been detected in human seminal plasma as well as on human ejaculated spermatozoa. This study aimed at studying the possible role of leptin in regulating human sperm functions. Immunofluorescent staining was used to study the expression of leptin and its receptor. The correlation between the concentration of leptin and soluble leptin receptor (ObRs) in seminal plasma as measured by enzyme-linked immunosorbant assay and sperm motility parameters measured by computer-assisted sperm analyais (CASA) was determined. The effects of recombinant leptin on human sperm motility, capacitation and acrosome reaction as measured by chlortetracycline staing were also studied. Leptin immunoreactivity was demonstrated at the equatorial and neck regions of human spermatozoa, whereas that of ObRs was shown up on the tail. After Percoll separation, spermatozoa with high density had more intense leptin immunoreactivity compared with those with low density. No significant correlation was found between seminal plasma concentration of leptin/ObRs and sperm motility parameters. After incubation with recombinant human leptin for either 3 h or overnight, there was no change in all the CASA motility parameters determined and percentages of capacitated and acrosome-reacted spermatozoa. We concluded that leptin does not have a significant effect on motility and capacitation/acrosome reaction in human ejaculated mature spermatozoa. Its role in male reproduction is yet to be determined.     

Assessment of released acrosin activity as a measurement of the sperm acrosome reaction

Aim： To develop a method for assessing sperm function by measuring released acrosin activity during the acrosome reaction （AR）. Methods： Human semen samples were obtained from 24 healthy donors with proven fertility after 3-7 days of sexual abstinence. After collection, samples were liquefied for 30 min at room temperature. Standard semen parameters were evaluated according to World Health Organization （WHO） criteria. Calcium ionophore A23187 and progesterone （P4） were used to stimulate the sperm to undergo AR. After treatment, sperm were incubated with the supravital dye Hoechst33258, fixed in a glutaraldehyde-phosphate-buffered saline solution, and the acrosomal status was determined by fluorescence microscopy with fluorescein isothiocyanate-labeled Pisum sativum agglutinin （FITC-PSA）. The percentage of sperm undergoing AR （AR%） was compared to sperm acrosin activities as assessed by spectrocolorimetry. The correlation between AR% and acrosin activity was determined by statistical analysis. Results： The AR% and released acrosin activity were both markedly increased with A23187 and P4 stimulation. Sperm motility and viability were significantly higher after stimulation with P4 versus stimulation with A23187 （P 〈 0.001）. There was a significant positive correlation between released acrosin activity and AR% determined by FITC-PSA staining （r= 0.916, P 〈 0.001）. Conclusion： Spectrocolorimetric measurement of released acrosin activity might serve as a reasonable alternative method to evaluate AR. （Asian J Androl 2008 Mar, 10： 236-242）     

Effects of tumour necrosis factor alpha and interleukin-6 on progesterone and calcium ionophore-induced acrosome reaction

For human spermatozoa to successfully fertilize the oocyte, they need to undergo a timely acrosome reaction (AR). Factors which disturb the AR may lead to fertilization failure. The objective of this study was to investigate the effects of two cytokines namely tumour necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) on the spontaneous, calcium ionophore-induced and progesterone-induced human sperm AR. Twenty-two normal semen samples were treated with increasing concentrations of TNF-α and IL-6 after spermatozoa were isolated by a double wash swim-up method. The AR was induced by calcium ionophore A23187 and progesterone. The AR was determined by using fluorescein isothiacyanate Pisum sativum agglutinin and observed under fluorescence microscope. Both TNF-α and IL-6 could decrease the spontaneous, ionophore and progesterone-induced AR ( p  <   0.05) in a dose-dependent manner. TNF-α showed a more potent inhibiting effect than IL-6 by inhibiting the AR at lower concentrations. This study has demonstrated that TNF-α and IL-6 play a role in inhibiting both the non-physiological as well as physiologically elicited AR by calcium ionophore and progesterone respectively.     

Relation of human sperm acrosin activity and fertilization in vitro

The aim of the study described here was to determine the possible contribution of the acrosin activity test to routine semen analysis in enhancing the precision of the prognosis of IVF success in a group of patients in which the contribution of the egg factor to infertility was ruled out (20 cases) compared to a control IVF group (39 cases). Semen analysis, acrosin activity and acrosome ultrastructure were determined for all semen samples. The group with high fertilization rates was comprised of normozoospermic patients while the group with low fertilization rates was comprised of astheno-teratozoospermic patients. The mean acrosin level of the positive IVF group was significantly higher than that of the negative group (51.7 ± 33.2 and 28.6 ±13.7, respectively). Two parameters: per cent motile spermatozoa and acrosin level, were found to have a significant positive correlation with subsequent successful IVF ( r = 0.36, P < 0.006; r = 0.37, P < 0.004, respectively); and agenesis of the acrosome was found to have a significant negative correlation ( r = -0.33, P < 0.01). The ability of these parameters to correctly predict fertilization success was 59%, with 5% false positive, among which 15.4% was predicted solely by the acrosin level (above 54 μIU 10⁶ cells⁻¹) and 23% solely by per cent motile spermatozoa (above 50%). Abnormalities of the acrosome ultrastructure did not contribute further to the correct classification. The apparent clinical benefit of the acrosin level test is discussed.     

Assessment of the effect of testosterone on the acrosome reaction of human spermatozoa

In the acrosome reaction, the spermatozoon plasma membrane fuses with the outer acrosomal membrane, resulting in the release of the acrosomal content. Several compounds, such as sex steroids, are known to modulate the acrosomal exocytosis. Testosterone regulates various functions in male reproductive physiology; however, little is known about the relationship between testosterone and the acrosome reaction. Thus, our objective was to study the effect of testosterone on the acrosome reaction of human spermatozoa. To evaluate the acrosomal exocytosis, spermatozoa were incubated with testosterone (0.2, 2.0 and 20 nmol l(-1)), progesterone and control medium for 60, 120, 240 and 1440 min. The acrosome reaction was assessed by staining with Hoechst 33258 and fluorescein isothiocyanate-conjugated P. sativum agglutinin lectin. In general, spermatozoa incubated with progesterone had the highest percentage of acrosomal exocytosis. The percentage of acrosome reaction obtained in the three treatments with testosterone differed from that observed for progesterone at 120, 240 and 1440 min (24 h). Additionally, significant differences were found between testosterone (2.0 and 20 nmol l(-1)) and progesterone after 60 min. Differences between control and the three testosterone treatments studied were obtained only at 1440 min. In general terms, these results show that testosterone exerts no inductor effects on the acrosome reaction of human spermatozoa.     

Effect of zinc on human sperm motility and the acrosome reaction

This study has assessed the effect of zinc on human sperm motility and the acrosome reaction in vitro. Progressively motile human sperm were selected by swim-up and by glass bead columns and then incubated in a medium in which capacitation happened in an asynchronous way. Different doses of zinc (1, 10, 100 and 1000 microM) were added for periods of 2, 4 or 6 h. Other samples were incubated with zinc (1000 microM), and after 1 h incubation, the zinc was removed. Aliquots of each culture were used to evaluate progressive motility and the acrosome reaction using a triple-stain technique. Sperm motility was reduced when the amount of zinc added was greater than or equal to 100 microM, and these doses also caused a significant reduction in the % of sperm undergoing the acrosome reaction. After removal of zinc and further incubation in zinc-free medium for 1 h, an increase in the percentage of motile and acrosome-reacted sperm was observed. However, the increase in acrosome reaction did not reach the values observed in controls. Results suggest that extracellular zinc acts as an inhibitor of human sperm motility and the acrosome reaction (and/or capacitation and the acrosome reaction). This inhibitory effect is reversible and occurs in a dose-dependent fashion. The probable mechanisms involved are discussed.