精浆内脂质运载蛋白型前列腺素D合成酶与α-葡萄糖苷酶的相关性研究

目的分析脂质运载蛋白型前列腺素D合成酶(L-PGDS)与精浆其他参数之间的关系,探讨L-PGDS在男性生殖系统中的作用。方法分析92份精液中的精子密度、精子活力、L-PGDS浓度、酸性磷酸酶活力以及α-葡萄糖苷酶活力。依据精子密度,将标本分为3组:正常组(精子密度>20×106/ml)、寡精子组(精子密度<20×106/ml)及无精子组(精子密度为0)。彩色精子质量分析系统测定精子密度及活力,双抗体夹心酶联免疫吸附试验(ELISA)检测精浆内的L-PGDS浓度,分光光度计测定α-葡萄糖苷酶活力。结果正常组、寡精子组以及无精子组患者精浆L-PGDS浓度依次降低,差异显著(P<0.001)。L-PGDS的浓度与α-葡萄糖苷酶、精子密度及精子活力呈正相关,相关系数(r)分别为0.426、0.813和0.380。结论精浆L-PGDS浓度可作为少精子症的辅助诊断指标。     

不育患者精浆α-1,4糖苷酶活性与精液参数的关系

目的　探讨精浆 α-1 ,4糖苷酶活性与精液参数之间的关系。　方法　分光光度比色法测定精浆 α-1 ,4糖苷酶活性及进行精液常规分析。　结果　 2 90 2例男性不育者精浆α-1 ,4糖苷酶活性异常率为 3 8.87%。该酶活性与精子密度、精子活率、a,b级精子活力和顶体酶活性呈显著正相关 ( r分别为 0 .460、0 .1 2 2、0 .0 86和 0 .2 3 0 ,P均 <0 .0 0 1 ) ,而与精液量、精液 p H、液化时间和畸形精子率无显著相关 ( P>0 .0 5)。 α-1 ,4糖苷酶活性正常组精子密度、活率、a,b级精子活力和精子顶体酶活性均明显高于 α-1 ,4糖苷酶活性异常组 ( P<0 .0 0 1 )。以常规精液分析法 ( RSA)主要参数正常与否分成的两组间α-1 ,4糖苷酶活性差异有显著性 ( P<0 .0 0 1 )。　结论　α-1 ,4糖苷酶活性对精子密度、活率、a,b级精子活力和顶体酶活性均有明显影响 ,对精液量、精液 p H、液化时间和畸形精子率无显著影响     

精浆尿酸的检测及其与精液参数的相关性研究

目的:建立精浆尿酸(UA)的检测方法并探讨精浆UA水平与精液参数的相关性。方法:根据检测血清UA的方法加以改良建立检测精浆UA的方法,并观察批内变异和不同技术人员检测结果之间的差异以评价方法的可接受性。同时分析138例男性精浆UA水平与患者年龄、禁欲时间、精液量、pH、精子密度、活动率、a+b级精子百分率和正常形态精子百分率之间的相关性。结果:精浆UA检测方法的批内变异为9.16%,2位技术人员的检测结果没有显著性差异(P=0.541)。精浆UA水平与正常形态精子百分率呈正相关(r=0.350,P=0.025),与精子活动率和a+b级精子百分率有呈正相关的趋势(r=0.147和0.156,P=0.085和0.068),而与精液分析其他参数如精液量、pH、精子密度、禁欲时间以及患者年龄等无相关性。结论:通过对血清UA检测方法加以改良可以建立可接受的精浆UA检测方法。精子形态学、活动率及a+b级精子百分率可能与精浆UA水平高低有关。     

Alpha-glucosidase activity and sperm motility

We correlated the activity of alpha-glucosidase in seminal plasma with the motility and differential motility of sperm. Significant positive correlations were found between the alpha-glucosidase activity and both motility and the percentage sperm with good forward progression. This supports the use of alpha-glucosidase in semen as a marker of epididymal function and specifically of the development of motility.     

Standardization and quality control for the determination of alpha-glucosidase in seminal plasma

We attempted to provide an instruction for the standardization of alpha-glucosidase level determination and quality controls within the same laboratory and among different laboratories. Each of 51 semen samples was divided into two aliquots, centrifuging at 1000 g for 10 min or 3000 g for 15 min. The alpha-glucosidase level and sperm concentration of each sample were assayed. The alpha-glucosidase level in seminal plasma obtained at 3000 g for 15 min centrifugation was significantly lower than that at 1000 g for 10 min (p = 0.001). An additional 6 samples of seminal plasma with or without phenylmethylsulfonyl fluoride (PMSF), obtained at 3000 g for 15 min centrifugation, were frozen for 20 days, and each of 6 samples was determined for their alpha-glucosidase levels after thawing every other day. There was no significant difference between alpha-glucosidase levels in seminal plasma regardless of the presence of PMSF. The alpha-glucosidase level increased with the length of abstinence period. In conclusion, centrifugal velocity and abstinence time should be standardized in the alpha-glucosidase determination. Frozen seminal plasma may serve as the sample for quality control among clinical laboratories.     

Relationships between seminal plasma arginase activity and spermatological parameters in rams

This study was conducted to evaluate the correlation between seminal plasma arginase activity and spermatological parameters in rams. In this study, five fertility-proven Awassi rams were used as material. Six ejaculates were collected from each ram by an artificial vagina. Spermatological parameters (semen volume, mass activity, sperm motility and concentration and abnormal sperm rate) were immediately determined in each ejaculate. For enzyme assay, the semen samples were centrifuged and stored at -20 °C for the analysis of arginase activity. The average seminal plasma arginase activity was 0.61 ± 0.20 U (mg protein)(-1) . There was a positive correlation between arginase activity and semen volume (r = 0.412, P < 0.05), semen mass activity (r = 0.610, P < 0.01), sperm motility (r = 0.447, P < 0.05) and sperm concentration (r = 0.808, P < 0.01). However, there was a negative correlation between arginase activity and abnormal sperm rate (r = -0.424, P < 0.05). In conclusion, this study clearly suggests that there is a significant correlation between seminal plasma arginase activity and spermatological parameters. In light of these results, seminal plasma arginase activity may be a biochemical criterion for determining sperm quality besides classical semen analysis parameters in rams.     

Relationship between semen quality and seminal plasma components: alpha-glucosidase, fructose and citrate in infertile men compared with a normospermic population of Tunisian men

The aim of this study was to evaluate the correlation between the secretory function of the male accessory glands and sperm parameters in normospermic controls and infertile patients. One hundred and fifty-nine men were investigated: they were composed of two groups: normospermic ( n  = 37) and infertile ( n  = 122) men with altered sperm characteristics. These infertile men were divided into the following groups: asthenozoospermia ( n  = 38), teratozoospermia ( n  = 40) and asthenoteratozoospermia ( n  = 44). The patients underwent semen analysis and measurements of fructose, neutral alpha-glucosidase and citric acid. The level of fructose was significantly decreased in asthenozoospermic and increased in asthenoteratozoospermic men. It was significantly correlated with semen volume, sperm count, motility and morphology. The seminal alpha-glucosidase levels were significantly correlated with semen volume and pH and citric acid was significantly correlated with pH. Thus, alpha-glucosidase and citric acid levels were associated with semen pH. The significant correlation between semen parameters, accessory glands and epididymal functions highlights the relationship between semen and normal genital tract function.     

精浆酸性磷酸酶和γ-L-谷氨酰转肽酶检测的比较及其与精液参数的相关性研究

目的:对精浆酸性磷酸酶(ACP)和γ-L-谷氨酰转肽酶(γ-GT)活性检测进行比较,并分析ACP和γ-GT活性与精液参数的相关性。方法:133例精浆标本,分别检测ACP活性和γ-GT活性。随机留取2例精浆标本,1例用于ACP批内检测,另1例用于γ-GT批内检测。随机留取4例标本,2例用于ACP批间检测,另2例用于γ-GT批间检测。用计算机辅助的精液分析(CASA)系统分析精液标本的精液量、pH、精子密度、活动率、a+b级活动精子百分率等参数。同时分析ACP和γ-GT活性与精液参数的相关性。结果:精浆ACP活性和γ-GT活性呈显著性正相关(r=0.570,P=0.000)。ACP的批内变异系数(CV)为13.72%,批间CV分别为13.80%和15.49%。γ-GT批内CV为7.68%,批间CV分别为7.76%和9.73%。精浆ACP活性和γ-GT活性均与pH值呈显著负相关(r=-0.330,P=0.000;r=-0.388,P=0.000)。γ-GT活性与精子密度呈显著正相关(r=0.165,P=0.045),而ACP活性与精子密度无显著相关(r=0.048,P=0.546)。ACP活性和γ-GT活性均与精子活动率、(a+b)级活动精子百分率、精液量、禁欲时间以及年龄无显著相关性。结论:精浆γ-GT活性检测的精确性高于ACP活性检测,两者与精液参数的相关性基本类似。提示精浆γ-GT活性检测比ACP活性检测更适合用来评价前列腺功能。     

Relationship between seminal ascorbic acid and sperm DNA integrity in infertile men

Ascorbic acid has recently been reported to protect sperm DNA from the damage induced by exogenous oxidative stress in vitro. But, there is no report on seminal ascorbic acid and sperm DNA fragmentation in infertile men. In this study, we asked whether sperm DNA damage correlates with seminal ascorbic acid levels. Sperm DNA fragmentation index (DFI) was analysed in 75 men by flow cytometry after acridine orange staining. We also measured the levels of seminal plasma ascorbic acid and total antioxidant capacity. Abnormal sperm DNA integrity (DFI >or= 30%) was observed in 12% of the patients with normal semen parameters and in 52% of the patients with abnormal semen parameters. There were significant correlations between the level of DFI and conventional semen parameters including sperm count, motility and morphology (r = -0.29, -0.55 and -0.53 respectively; p < 0.05). Seminal ascorbic acid level was significantly lower in the patients with leucospermia than the patient with normal semen parameters. Interestingly, a significantly greater percentage of men with abnormal DFI were observed in the patients with low levels of seminal ascorbic acid compared with those with normal or high levels of ascorbic acid (59% vs. 33%, p < 0.05). Men with insufficient seminal ascorbic acid frequently have sperm DNA damage.     

Visco-elasticity of seminal fluid in relation to the epididymal and accessory sex gland function and its impact on sperm motility

Seminal viscopathy was shown to be associated with male infertility. However, our knowledge about the regulatory mechanism of this process is still limited. In semen samples from 411 men attending for fertility assessment, traditional semen parameters including visco-elasticity were assessed according to the World Health Organization guidelines. Sperm motility was evaluated by use of computer aided sperm analysis (CASA). Seminal activity of neutral alpha-glucosidase (NAG) and concentrations of prostate-specific antigen (PSA), zinc, and fructose were measured. The activity of NAG, and the concentrations of PSA and zinc were significantly lower in hyper-visco-elastic semen samples (medians: 5 vs. 8 mU/mL; 741 vs. 924 mg/L; 1 vs. 2 mM/L), than in those with normal visco-elasticity (p = 0.004, 0.005 and 0.011, respectively). When comparing the total amounts, only for seminal fructose there was a difference between samples with high visco-elasticity as compared with those of normal visco-elasticity (median: 74 vs. 53 microM/ejaculate, p = 0.007) This seminal marker was the only significant independent parameter in predicting seminal visco-elasticity in a multiple logistic regression analysis (odds ratio for the highest quartile = 4.67). Hyper-visco-elasticity was associated with a lower percentage of motile spermatozoa (43 vs. 50%, p = 0.045). Similar trend was found for the CASA motility characteristics curvilinear velocity (VCL), average path length (VAP), amplitude of lateral head displacement (ALH) (p = 0.008, 0.038 and 0.020, respectively). Our study demonstrated the interplay between the regulatory effect of post-testicular organs on semen visco-elasticity. Hyper-visco-elasticity was associated with asthenozoospermia and lower levels of VCL, VAP and ALH.     

Why do we determine α-glucosidase activity in human semen during infertility work-up?

alpha-Glucosidase is a normal constituent of human semen, produced mainly in the epididymis. It is significantly correlated to sperm count. Its activity is low in cases of epididymal obstruction. We evaluated alpha-glucosidase activity in 653 semen samples of patients, who attended our department for marital infertility, with respect to associations to clinical and other seminal parameters. The normal range (mean +/- 2 SD) in samples with normal parameter values was 7.2-46.4 mU ml-1. The determination in patients with azoospermia revealed mean values of 7.7 +/- 9.5 mU ml-1 in obstructive azoospermia, and 15.8 +/- 11.5 mU ml-1 in nonobstructive azoospermia. The difference was not statistically significant in that the sensitivity of determination with respect to the presence of obstruction was only 0.66, and the specificity 0.83. A significant correlation (r = 0.34) of alpha-glucosidase activity with log sperm count was observed. The mean alpha-glucosidase activity was not significantly different in groups formed according to sperm motility, according to leucocyte count or according to semen volume. A difference between smokers and nonsmokers with comparable sperm count, as reported in the literature, did not occur. We conclude from our results that the determination of alpha-glucosidase activity does not give additional information of the fertility status exceeding that of other clinical investigations or parameters of semen analysis.     

Seminal plasma albumin: origin and relation to the male reproductive parameters

We wanted to investigate the origin of seminal plasma albumin and its relation to the male reproductive parameters. Semen samples from 916 men, under infertility assessment, were analysed according to guidelines of the World Health Organization. Seminal plasma constituents, i.e. albumin, markers of the epididymal (neutral alpha-glucosidase, NAG), prostatic (prostate-specific antigen, PSA, and zinc) and seminal vesicle function (fructose), as well as levels of reproductive hormones in plasma were measured. The sperm chromatin structure assay (SCSA) was applied on 267 of the 916 samples. A negative correlation was seen for seminal albumin and plasma follicle-stimulating hormone (r=-0.1, P=0.02) and a positive correlation for seminal albumin and serum inhibin B (r=0.2, P=0.004). Albumin exhibited positive correlations with the epididymal marker, NAG (r=0.5, P<0.001) and with the prostatic markers, PSA and zinc (r=0.1, P=0.001; r=0.2, P<0.001 respectively) as well as with age (r=0.2, P<0.001). A negative significant association was seen for seminal albumin and semen volume (beta=-0.60; 95% CI -0.80 to -0.30). The opposite trend was found regarding sperm concentration (beta=0.34; 95% CI 0.30-0.40), total sperm count (beta=0.30; 95% CI 0.20-0.40), and percentage morphologically normal spermatozoa (beta=0.70; 95% CI 0.10-1.0). No association was found between albumin and sperm motility, SCSA parameters, or fructose, the marker of seminal vesicles. Our results suggest testicular, epididymal and prostatic origin of seminal plasma albumin, in addition to the contribution from blood. This is the first study to demonstrate an association between seminal plasma albumin and sperm morphology. Further studies are needed to elucidate the role of seminal albumin in sperm morphology.     

精浆游离L-肉毒碱水平及其与精子密度、活动率及活力的相关性研究

目的:研究正常生育及不育男性精浆中游离L-肉毒碱水平差异及其与精子密度、活动率(a+b+c级精子百分率)及活力(a+b级精子百分率)之间的相关性,探讨精浆中游离L-肉毒碱水平对男性生育力的影响及其在不育症检查和治疗中的作用。方法:分别采用高效液相色谱法和计算机辅助精液分析系统,测定了230例不育症患者(精子密度正常117例,少精子症81例,无精子症32例)和30例正常生育男性精浆中游离L-肉毒碱水平及精子密度、活动率、活力等参数。根据检查结果对不育症患者分组后,以SPSS12.0软件包进行统计学分析,比较各组间游离L-肉毒碱水平的差异以及游离L-肉毒碱水平与精子密度、活动率、活力之间的相关性。结果:正常生育组精浆游离L-肉毒碱水平明显高于不育组(P<0.01)。精液中精子密度越低、活力越弱,这种差异性越显著。相关性分析结果显示,精浆游离L-肉毒碱水平与精子密度呈显著正相关关系(r=0.521,P<0.01),与精子活动率和活力之间也具有正相关关系(r=0.319,P<0.01;r=0.251,P<0.01)。结论:精浆L-肉毒碱水平与精子密度、活动率和活力之间密切相关,其含量测定作为一项有用的生化指标,可为男性不育症检查及临床诊治和进行有关男性生殖功能机制研究提供参考。     

Smoking influence on sperm vitality,DNA fragmentation,reactive oxygen species and zinc in oligoasthenoteratozoospermic men with varicocele

This study aimed to assess the influence of smoking duration and intensity on sperm vitality, sperm DNA fragmentation, reactive oxygen species (ROS) and zinc (Zn) levels in oligoasthenoteratozoospermic (OAT) men with varicocele (Vx). A total of 246 men were investigated who were divided into OAT nonsmokers, OAT smokers, OAT nonsmokers and OAT smokers with Vx. They were subjected to history taking, clinical examination and semen analysis. In their semen, sperm hypo‐osmotic swelling (HOS) test, sperm DNA fragmentation test, seminal ROS and seminal Zn were assessed. The results demonstrated significantly decreased HOS test, seminal Zn level and significantly increased sperm DNA fragmentation, seminal ROS levels in OAT smokers with Vx more than OAT smokers compared with OAT nonsmokers. Smoking intensity, smoking duration and Vx grade demonstrated significant negative correlations with sperm motility, HOS test percentage and significant positive correlations with sperm DNA fragmentation, seminal ROS level. It is concluded that smoking has a negative impact on sperm progressive motility, HOS test, seminal Zn and positive impact on sperm DNA fragmentation, semen ROS level that are exaggerated if Vx is associated being correlated with smoking intensity, smoking duration and Vx grade.     

Seminal immunoreactive relaxin in domestic animals and its relationship to sperm motility as a possible index for predicting the fertilizing ability of sires

Although immunoassayable relaxin has been detected in human and boar seminal plasma, there is no evidence suggesting the existence of immunoreactive relaxin in the seminal plasma of other domestic animals. The first objective of this study was to determine whether immunoreactive relaxin was present in the seminal plasma of bulls, rams and he-goats. In addition, the correlation of immunoreactive relaxin with sperm motility as an index for predicting the fertilizing ability of bull sires was investigated. Semen with normal sperm motility was collected from bulls, rams and he-goats, and the relaxin immunoreactivity of the semen samples was measured using a time-resolved fluoroimmunoassay (TR-FIA) for porcine relaxin that we developed. The presence of relaxin immunoreactivity was demonstrated in seminal plasma from bulls, rams and he-goats. The level of immunoreactive relaxin in seminal plasma was the highest in bulls followed by humans, rams, boars and he-goats in that order, when relaxin levels in boar and human semen having normal sperm motility were also assayed under the same conditions. When the correlation between the seminal plasma level of immunoreactive relaxin and sperm motility was examined in bull semen samples as an index for predicting fertilizing ability, it was found that the relaxin level was significantly correlated with the percentage of spermatozoa showing the most intensive motility (r = 0.64, p < 0.05). These results indicate that immunoreactive relaxin is widely found in the seminal plasma of domestic animals and that measuring the relaxin concentration of seminal plasma may be useful to identify subfertile sires or predict the fertility potential of individual sires.     

Seasonal changes of neutral alpha-glucosidase activity in human semen

Although there are contradictory reports, the biochemical evaluation of the seminal activity of neutral alpha-glucosidase (NAG) has repeatedly been described as an important parameter to test epididymal patency and is recommended by the WHO. Because, for a number of diagnostic parameters, seasonal variations have been described even in the human, it was the aim of this study to investigate possible circannual changes of NAG. This is an important aspect of andrological diagnosis, as seasonal changes of specific diagnostic parameters might have an impact on the accuracy and predictive power of these parameters, which in turn might have an effect on the therapeutic concept for the patients. In a total of 473 patients, sperm concentration, volume of the ejaculate, total motility, progressive motility, pH value, number of peroxidase-positive cells, concentration of fructose, and NAG as functional markers of the seminal vesicles and the epididymis, respectively, were analyzed according to standard procedures. Seminal activity of NAG was significantly correlated with the sperm concentration (P < .0001), ejaculate volume (P < .0001), and the pH (P = .0025). Moreover, significant (P = .0008) seasonal changes in the activity of seminal NAG with the maximum in spring (76.87 mU/ejaculate) and the minimum in autumn (58.55 mU/ejaculate) were found. The incidence of low-ranged activity of the enzyme was 9.2% in spring, while it was 20.3% in autumn. Thus, our data clearly demonstrate circannual changes of the seminal activity of neutral alpha-glucosidase. This in turn has clinical impact as the predictive power of the test system changes throughout the year.     

Sperm characteristics and accessory sex gland functions in HIV-infected men

A comparative study was carried out in the andrology clinic, Parirenyatwa Hospital, Harare, Zimbabwe, to investigate the sperm characteristics and accessory sex gland functions in HIV-infected individuals. Sixty-two patients with infertility problems who attended the clinic were requested to donate semen and blood after consent was obtained. HIV antibodies in paired semen and blood samples, sperm morphology, sperm count, sperm motility, seminal leucocytes, seminal fructose, seminal neutral alpha-glucosidase, and citric acid were analyzed. Nine out of 31 blood samples tested positive, while 21 out of 62 semen samples were positive for HIV. Leucocytospermia was associated with HIV-seropositive men (p < .01). The accessory sex gland function, as evaluated by biochemical markers, was not affected in HIV-seropositive men. HIV causes impairment of sperm motility by activating seminal leucocytes, which in turn induce oxidative stress on the sperm. Leucocytospermia is almost always present in HIV-seropositive men.     

Usefulness of sperm quality analyzer-V (SQA-V) for the assessment of sperm quality in infertile men

The aim of this study was to evaluate the correlation of new of Sperm Quality Analyzer (SQA-V) with the computer-assisted sperm analysis (CASA) and manual semen analysis estimates. One hundred five fresh semen samples were analyzed using SQA-V and CASA and manual semen analysis. Significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and normal morphology (p < 0.0001) were observed between SQA-V variables and manual semen analysis estimates. There also were significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and sperm velocity (p = 0.0235) between SQA-V variables and CASA estimates. Meanwhile, it did not correlate with amplitude of lateral head displacement, beat cross frequency, lineality assessed by CASA. The value of the sperm concentration and the sperm motility measured by SQA-V showed high correlations with the value of those measured by CASA and manual semen analysis. In addition, velocity and sperm morphology may also be evaluated to some extent using SQA-V.     

不育男性精浆总抗氧化能力与精子运动功能的关系

目的:研究不育男性精浆总抗氧化能力(TAC)与精子运动能力和方式之间的关系,探讨精浆TAC水平在男性生育中的临床意义。方法:113例精子密度正常的不育男性,28例正常生育男性作为对照组。精液于37℃液化后采用计算机辅助精液分析(CASA)系统进行精液常规分析,采用比色法进行精浆TAC分析。结果:正常生育组精浆TAC为(19.82±6.33)U,不育男性精子密度正常组精浆TAC为(14.37±8.45)U,不育男性精子密度正常组与正常生育组比较存在显著性差异(P<0.01)。精浆TAC与a级精子百分率(r=0.208,P<0.05)和(a+b)级精子百分率(r=0.231,P<0.05)呈显著正相关,精浆TAC与精子运动参数中的前向性(r=0.200,P<0.05)、直线性(r=0.208,P<0.05)、曲线速度(r=0.189,P<0.05)、直线速度(r=0.210,P<0.05)、平均移动速度(r=0.215,P<0.05)及鞭打频率(r=-0.248,P<0.01)之间有显著的相关性,其中前向性、直线性、直线速度、曲线速度、平均移动速度与TAC呈正相关(P<0.05),而鞭打频率与TAC呈负相关(P<0.01)。精浆TAC与摆动性、侧摆幅度、平均移动角度之间无显著相关。结论:精浆中TAC水平与精子运动能力和运动方式密切相关,适宜的精浆TAC为精子运动提供了良好的外部环境,精浆中过低的TAC水平与精子运动能力下降和运动方式改变有关,可能是引起男性不育的病因之一。精浆中TAC分析可为探讨男性不育的发病机制以及临床用药提供依据。     

alpha-Glucosidase, sperm ATP concentrations, and epididymal function

It has been suggested that alpha-glucosidase may be a marker of epididymal patency and function. Spermatozoal ATP concentrations decrease during passage through the epididymis, indicating efficient maturation. We correlated sperm motility with seminal plasma alpha-glucosidase activity and spermatozoal ATP. The sperm motility correlation with alpha-glucosidase activity was significantly positive, and the sperm motility correlation with spermatozoal ATP was significantly negative. It appears that high-alpha-glucosidase activity and low-spermatozoal ATP were present in semen with good sperm motility and could possibly indicate efficient epididymal function.