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。     

南德士抑制人精子顶体酶活性的实验研究

目的:研究顶体酶抑制剂南德士对人精子顶体酶的抑制作用。方法:利用10例已生育的健康男性精子,分别与0.100、0.120、0.144、0.173、0.207、0.249、0.299、0.358、0.430mmol/L浓度的南德士在30℃条件下孵育5min,通过改良的Kennedy法检测其残余顶体酶活性。对照组使用经典的顶体酶抑制剂盐酸-N2-对甲苯磺酰-L-赖氨酸甲基甲酮(TLCK),其浓度分别是150.0、189.8、213.6、240.3、270.3、304.1、342.1mmol/L。结果:残余顶体酶活性与南德士的浓度呈负相关(r=-0.9881);南德士对顶体酶的抑制能力比TLCK高近800倍。结论:南德士能高效抑制人精子顶体酶活性。     

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。     

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

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

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。     

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     

影响顶体酶活性相关因素的研究进展

精子顶体酶存在于精子顶体内，是一种胰蛋白酶，它以酶原形式合成并储存在顶体内，它是受精过程中一种重要的蛋白水解酶。当精卵结合发生顶体反应时，通过顶体酶原的释放，为精卵结合提供了条件。顶体酶活性低下会影响卵细胞卵丘的分解及精子对卵细胞透明带的穿透，从而导致男子不育。目前，计算机辅助精子分析（CASA）系统已经成为临床上评价男性生育能力的主要方法，但该方法在检测方面尚有一定的主观性和局限性，对全面了解精子生育能力尚显不足。许多资料表明顶体酶活性是判断精子生育力一项很有价值的指标，特别是对不明原因的男子不育者。本文对国内外近年来与顶体酶活性研究相关的文献进行了整理并综述如下。     

IL-6对人精子顶体反应影响机制的初步探讨

目的:探讨IL-6对人精子顶体反应(AR)的影响机制。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:IL-6可诱导精子顶体酶及超氧化物歧化酶(SOD)的活性,促进精子顶体反应;胞外Ca2+单独不能诱导精子顶体反应,且没有胞外Ca2+的参与,IL-6也不能诱导精子顶体反应;蛋白激酶C(PKC)抑制剂calphC能逆转IL-6诱导的精子顶体反应。结论:IL-6对精子顶体反应有一定的促进作用,可能通过诱导精子的顶体酶和SOD活性等途径来实现,在此作用中,也涉及了PKC的激活,且还需要外源性Ca2+的参与。     

优选前后精子顶体酶活性与IVF受精率相关性的研究

目的探讨优选处理前和处理后精子顶体酶活性的变化,及与体外受精(IVF)受精率的相关性。方法采用分光光度比色法,对接受IVF治疗的53例不育夫妇男方精液,分别测定优选处理前和处理后精子顶体酶活性,分析其与IVF受精率的相关性。结果优选处理后精子顶体酶活性与优选前比较有显著性差异(P<0.05);达到常规IVF标准并选择常规IVF治疗者,优选后的精子顶体酶活性与IVF受精率有相关性,精子顶体酶活性降低与IVF受精率降低有关。结论精子顶体酶活性与IVF受精率有相关性,并且通过精子顶体酶活性可以预测IVF受精率。     

KF-950对人精子顶体素活性及顶体的影响

目的 :研究顶体素抑制剂 4 胍基苯甲酸盐 (KF 95 0 )对人精子顶体素的抑制作用和对顶体的影响。　方法 :醋酸抽提并纯化人精子顶体素 ,BAEE/ADH法测定不同剂量KF 95 0作用后残余酶的活性。以生物素标记的豌豆凝集素为探针 ,采用ABC染色法观察KF 95 0对完整精子顶体的影响。　结果 :①正常人精子顶体素活性为(37.6 5± 4 .4 7)U/L(pH 8.5 ,2 5℃ ) ;②残余酶活性与KF 95 0剂量呈负相关 (r =- 0 .998) ,并呈量 效依赖关系 ,回归方程 :Y =7.5 7- 1.895X ;③随药物浓度的增加和作用时间的延长 ,顶体完整着色率均明显下降 (P <0 .0 1)。结论 :KF 95 0能直接抑制顶体素活性 ,推测对精子顶体也有明显的破坏作用 ,是一种新型、高效的精子顶体素抑制剂。     

Demonstration of an Anionic Acrosin Inhibitor in Spermatozoa Epididymal Fluid and Seminal Plasma of the Boar

A new anionic acrosin inhibitor was found in an acidic extract of boar spermatozoa. The protein was purified by hydrophobic and ion exchange chromatography. According to gel filtration and SDS-electrophoresis the inhibitor preparation shows a molecular mass of Mr approximately 6000-7000 daltons. The isoelectric point is close to pH 4.5. It is an effective inhibitor of boar acrosin and bovine trypsin, but it does not inhibit porcine plasmin and pancreatic kallikrein or bovine chymotrypsin. An inhibitor with identical properties was found in high concentration (97% of the total acrosin inhibiting activity) in the fluid of cauda epididymis and also as a minor acrosin inhibiting component (2% of total acrosin inhibiting activity) in seminal plasma. The results indicate that binding of the inhibitor to spermatozoa may have taken place in the epididymis.     

A simple, clinical assay to evaluate the acrosin activity of human spermatozoa

Acrosin, a sperm-specific acrosomal proteinase, has an essential role in the fertilization process. Low levels of acrosin appear to be associated with subfertility and infertility, and the acrosin activity of spermatozoa may potentially be a useful indicator of semen quality. The standard acrosin tests employed by research laboratories are too complicated and/or time consuming for clinical use; therefore, a simple assay has been developed to assess total acrosin activity (acrosin and activatable proacrosin). To perform the test, liquefied semen is centrifuged over Ficoll, the washed sperm pellet is suspended in a detergent (Triton X-100)-substrate (N-alpha-benzoyl-DL-arginine p-nitroanilide) buffer, pH. 8.0, and the amidase activity is determined spectrophotometrically after a 3-hour incubation period. Amidase activity can be inhibited with benzamidine, indicating that the activity is primarily or entirely due to acrosin. The absence of detergent in the incubation medium results in greatly reduced activity. The assay is repeatable, linear with increasing sperm concentration, sensitive to a lower limit of 2 x 10(6) spermatozoa, and the results correspond to those obtained with a standard acrosin extraction and assay technique. Storage of ejaculates at 3 to 6 C or at 22 to 24 C for 24 hours does not affect the acrosin activity significantly but much higher temperatures can cause a loss of activity. Freezing ejaculates results in a large decrease in sperm acrosin activity. Leukocytes show minimal activity in the assay. Sperm populations prepared by a swim-up procedure average approximately a 2-fold higher acrosin activity than the original ejaculates. Preliminary experiments indicate that the average sperm acrosin activity of ejaculates whose spermatozoa successfully fertilize human eggs in vitro is significantly higher than those that do not fertilize eggs.     

Determination of sperm acrosin activity for evaluation of male fertility

Aim: To investigate a simple method for assaying acrosin activity for the evaluation of male fertility. Methods:The acrosin activity of 7.5 × 10~6 sperm without seminal plasma and acrosin activity inhibitors was assayed using N-α-benzoyl-DL-arginine-p-nitroanilide (BAPNA) and detergent (Triton X-100) as substrate. Results: The acrosin ac-tivity of 60 normal fertile men (35 ± 10 μIU/10~6 sperm ) was higher than that of 168 infertile men ( 16 ± 8 μIU/10~6sperm) (P <0.01). It was indicated that there was a significant positive correlation between the acrosin activity andthe sperm motility ( r ≥ 0.6534, P < 0.01) and a significant negative correlation between the sperm malformed rateand the WBC number ( r ≤ -0.5426, P < 0.01). The temperature and time of incubation and the sperm concentrationcould influence the assay results. Conclusion: Acrosin activity is an important index for the evaluation of male fer-tility. The approach developed by the authors is a simple method for the determination of acro     

精子形态率、顶体酶活性对IVF受精失败的影响

目的 探讨精液中精子形态率及顶体酶活性与IVF受精失败的关系.方法 回顾性分析2014年在本中心行IVF新鲜周期的982个周期患者,根据精子形态分析结果,将正常精子形态百分率(x)分为5组,包括:1组:x≥4％,2组:x＜1％,3组:1％≤x＜2％,4组:2％≤x＜3％,5组:3％≤x＜4％.根据顶体酶活性分为正常组及异常组.结果 IVF受精失败组(受精率＜30％)与正常组比较,正常精子形态率及顶体酶活性差异有统计学意义(P＜0.01).精子形态率比较,2、3、4组与1组比较,差异有统计学意义(P＜0.01).5组与1组比较,差异无明显统计学意义(P＞0.05).顶体酶活性正常组与异常组比较,受精失败率差异有统计学意义(P＜0.01).结论 精子形态率及顶体酶活性异常可导致IVF受精失败率升高.     

On the Teratogenesis of Round-Headed Spermatozoa: Investigations with Antibodies Against Acrosin, an Intraacrosomally Located Acrosin-Inhibitor, and the Outer Acrosomal Membrane

Acrosin, the outer acrosomal membrane (OAM) and an acrosin inhibitor were studied in testicular cells and ejaculated spermatozoa of fertile men and in those of an infertile patient with exclusively round-headed spermatozoa in his ejaculates. The investigations were performed with the aid of immunohistochemical techniques using specific antibodies against the three acrosomal markers isolated from boar spermatozoa. The spermatozoa of fertile men exhibit staining for acrosin, the OAM and the acrosin inhibitor in the cap region while the round-headed spermatozoa of the patient are totally negative for the three markers, clearly supporting the conclusions of other authors that round-headed spermatozoa lack acrosomes. The lack of the acrosin system was further substantiated by the gelatin substrate film technique. In the course of normal human spermatogenesis acrosin, the OAM and the acrosin inhibitor we first demonstrable in early round spermatids, namely in identical compartments adjacent to the cell nucleus. During spermatid differentiation the staining for the three markers becomes flattened over the nucleus, resulting in a cap-like structure in testicular spermatozoa. In contrast to the ejaculated round-headed spermatozoa, the early round spermatids in the testis of the infertile patient exhibit fluorescent staining for the three markers in the region adjacent to the nuclear membrane. In the course of further spermiogenesis, the staining did not extend over the nuclear membrane, as was observed during normal spermiogenesis, but became separated from the nuclear membrane, as was observed during normal spermiogenesis, but became separated from the nuclear membrane, was translocated at various locations in the cytoplasm and was finally eliminated with the loss of the cytoplasm. These results are in accordance with the results of electron microscopically investigations on the teratogenesis of round-headed spermatozoa. Furthermore, the developmental pattern of the acrosin inhibitor during normal and abnormal spermiogenesis supports the intraacrosomal location of the acrosin inhibitor recently described by Tschesche et al. (1982).     

A serine protease inhibitor, N-alpha-tosyl-l-lysine chloromethyl ketone, prolongs rat hepatic allograft survival

BACKGROUND: Serine protease inhibitors have profound suppressive effects on cellular and humoral immune responses. We investigated the effect of a serine protease inhibitor, N-alpha-tosyl-l-lysine chloromethyl ketone (TLCK), on hepatic allograft survival in rats. Methods. Orthotopic hepatic transplantation was performed in an ACI (RT1(a))-to-LEW (RT1(1)) rat combination. TLCK was administered continuously at a dose of 4.4 mg/kg/day using an osmotic subcutaneous infusion minipump. RESULTS: TLCK prolonged hepatic allograft survival. Histologic staging of acute rejection based on Banff criteria in TLCK-treated hepatic allografts was significantly lower than in untreated allografts. TLCK significantly reduced serum concentrations of interferon (IFN)-gamma and tumor necrosis factor (TNF) alpha in allograft recipients. TNF-alpha mRNA levels in TLCK-treated allografts were significantly lower than in untreated allografts. TLCK also decreased perforin mRNA levels in hepatic allografts. Hepatic infiltrates eluted from TLCK-treated allografts showed significantly lower cell-mediated lympholytic activity against donor Con A blast cervical lymph node cells than those from untreated allografts. In vitro, TLCK suppressed interleukin-2 production and [(3)H]thymidine incorporation into an allogeneic mixed lymphocyte reaction. CONCLUSION: TLCK suppressed acute allograft rejection, suggesting a novel immunosuppressive strategy for therapy of acute organ rejection.     

Bradykininase and protease inhibitors in seminal plasma of fertile and infertile men.

The level of protease acrosin in the seminal plasma of oligozoospermic men was significantly higher than that of normozoospermic men. The amount of bradykininase in the seminal plasma was very high in both normozoospermic and oligozoospermic patients. When the acrosin and kininase content was referred to one million spermatozoa, seminal plasma kininase was significantly enhanced in oligozoospermic men, while the acrosin activity was similar in normozoospermic fertile men and infertile men. Human seminal plasma inhibitor I (HUSI I) increased along with sperm count. Human seminal plasma inhibitor II (HUSI II) showed no change. The motility of spermatozoa was depressed in oligozoospermic patients.     

Characterization of mouse epididymal acrosin: comparative studies with acrosin from boar and human ejaculated spermatozoa

Acrosin is an acrosomal protease believed to play a major role in fertilization. It is synthesized as an inactive precursor, proacrosin, which is processed via (auto)proteolysis into active form(s). In this paper, comparative studies on the characteristics of acrosin from mouse, boar and human are reported. The mouse proacrosin/acrosin was especially investigated to clarify whether or not the enzyme undergoes modifications during epididymal maturation. Acrosomal extracts from mature and immature mouse spermatozoa, as well as from ejaculated boar and human spermatozoa, were analysed by means of SDS–electrophoresis, Western blot and activity measurements. The studies showed that epididymal maturation produced a shift in the molecular weight of proacrosin. It was also observed that the activation kinetics differ strongly between the three species studied. Human proacrosin showed a constant substrate turnover, acrosin from boar showed sigmoidal activation kinetics and mouse acrosin, either from the caput or the cauda epididymides, showed a rapid decay in activity, suggesting the presence of an endogenous specific inhibitor.