精子顶体酶活性检测对诊断男性不育症的价值

目的比较各种质量精液的精子顶体酶活性。方法分光光度比色法检测顶体酶活性。结果各种质量精液的精子顶体酶活性均显著低于对照组P<0.001。结论精子顶体酶可作为了解精子生育力的一项可靠指标。     

Assessment of sperm functions in infertile patients with varicoceles

Acrosin activity, acrosome reaction and nuclear chromatin condensation were studied in 24 infertile patients with varicoceles and 26 fertile men with or without varicocele. Chromatin condensation, assessed by aniline blue staining, and acrosin activity, evaluated by gelatinolysis technique, were significantly affected in the group of infertile patients. Defective chromatin condensation and defective acrosin activity were detected in 67% and 50% of the infertile patients, respectively. No significant difference was found between the two groups regarding the acrosome reaction, which was assessed by the triple staining technique after exposure of spermatozoa to low temperature (4 degrees C). This study identified a subgroup of infertile patients with normal standard semen parameters but impaired sperm functions. Results of the sperm function tests and standard semen parameters were not correlated. Therefore, it is concluded from this study that important sperm functions are impaired in patients with varicocele and that the gelatinolysis technique and aniline blue staining are effective tools for assessment of the fertilization potential of varicocele patients.     

冷冻方法和冷冻保护剂对人精子顶体酶活性影响

目的:了解冷冻方法和冷冻保护剂对人精子顶体酶活性影响。方法:对精液标本进行速冻和缓慢冷冻保存,应用分光光度比色法测定精子顶体酶活性。结果:速缓冷冻法对精子顶体酶活性无显著性影响(P>0.05)。甘油-卵黄-柠檬酸钠(GYC)组顶体酶活性明显高于单一甘油组。结论:冷冻速度对人精子顶体酶活性没有影响(P<0.05),应用GYC作为CPM可使人精子顶体酶活性保持很高活性。     

禁欲时间对人精子顶体酶活性精子密度和精子数的影响

目的:了解禁欲时间对人精子顶体酶活性、精子密度和精子数的影响。方法:分光光度比色法测定精子顶体酶活性。结果:禁欲1～5d时,顶体酶活性是稳定的,而禁欲10d时顶体酶活性明显下降,与禁欲1d时顶体酶活性比较差异有显著性(P<0.05)。精子密度和精子数随禁欲时间增加而增加,禁欲10d精子密度约为禁欲1d精子密度2倍,禁欲10d精子总数约为禁欲1d的4倍。结论:禁欲时间影响顶体酶活性、精子密度和精子数。     

蛋白C抑制剂对人精子顶体反应和顶体素活性的影响

本研究应用提纯的尿PCI 进行对精子顶体反应和顶体酶的抑制试验。当洗涤后的精子与PCI 一起在体外孵育,其活动率和顶体反应未见降低;但当精子提取物或猪纯顶体素与PCI一起孵育时,其酰胺酶活性明显受到抑制且与PCI 剂量反应时间相关。这种抑制效应可能是通过形成PCI-顶体素复合物而起作用的,尽管这种复合物对SDS不稳定。精液中PCI 的高浓度(220±41.0mg/l)可能通过抑制由于精于变性或受损时释放出的顶体素或其它蛋白酶活性,从而保护生殖道上皮或精子本身不受酶的降解。     

Low Acrosin Activity in Polyzoospermia

Compared to normozoospermic men total sperm acrosin activity is considerably decreased (-59,1%) in patients with polyzoospermia indicating a functional defect of the proteolytic potential of the sperm acrosome, which may be responsible for the reduced fertility observed in this group of patients.     

精子顶体酶活性检测新方法与临床应用

以Na-苯甲酰-DL-精氨酸P-硝酰基苯胺(BAPNA)为底物,检测7.5×10~6去精浆精子顶体酶活性,孵育温度24℃,时间3小时,探讨和分析了最佳实验条件及注意事项.与此同时,检测了18例正常生育者精子(A组),16例精液常规基本正常者精子(B组),18例精液常规不正常者精子(C组).各组检测结果分别是29.7±14.3,19.4±10.9,6.9±7.7μIU/10~6精子(X±S),组间比较差异显著(P<0.05,P<0.001),提示顶体酶活性降低与生育力低下或不育相关.     

Ⅳ型前列腺炎对精子顶体酶活性及精液参数影响的研究

目的:探讨Ⅳ型前列腺炎对精子顶体酶活性及精液参数的影响。方法:对就诊的男性不育患者行精子顶体酶、精液参数及前列腺炎的相关检查。随机选择患有Ⅳ型前列腺炎的男性不育患者112例作为研究组,无Ⅳ型前列腺炎的男性不育患者123例作为对照组,比较两组之间精子顶体酶活性及精液参数的差别。结果:患有Ⅳ型前列腺炎的男性不育患者精子顶体酶活性、精子总活力、前向运动精子百分率、正常形态率及精子低渗肿胀试验与无Ⅳ型前列腺炎的男性不育患者相比,差异有显著性(P<0.05);而精液pH值、精子浓度,两组相比差异无显著性(P>0.05)。结论:Ⅳ型前列腺炎对男性不育患者的精子顶体酶活性、精子总活力、前向运动精子百分率、正常形态率及精子低渗肿胀试验等精液参数有影响。     

Acid extraction of acrosin from human spermatozoa pretreated by different physicochemical methods

Summary Washed human spermatozoa were subjected to different physicochemical methods, followed by acid extraction of the sperm acrosomes to dissolve acrosin. The acrosin activity of the sperm pellet and the supernatant was determined by benzoyl arginine ethyl ester/alcohol dehydrogenase (BAEE/ADH) assay to calculate the total acrosin activity in m U/10⁶ spermatozoa. Membrane-active and zymogen-activating agents increased the total acrosin activity 50%–200% compared to acid extraction alone. Similar results were obtained by osmotic shock, sonication and treatment with glass beads. Snap freezing of unprotected spermatozoa in liquid nitrogen yielded a fivefold increase in total acrosin activity, thus demonstrating that this is the method of choice for optimal acrosin extraction. The possibility is discussed as to whether acrosomal membrane alterations with improved solubilization of membranebound acrosin and/or conformational changes and/or zymogen activation are responsible for the considerable increase observed in acrosin activity.Dedicated to Prof. Dr. Dr. h.c. Otto Braun-Falco in honour of his 60th birthday     

精子质量与短时受精后受精结局关系的研究

目的 探讨精子质量对短时受精后受精结局的预测价值.方法 选取2009年1月至2010年6月在河南省人民医院生殖医学中心进行早期受精观察的常规体外受精胚胎移植(IVF-ET)的患者,筛除年龄＞ 38岁,且第2次减数分裂中期(MⅡ)卵母细胞数＜3个的患者,共有558个周期.按是否进行补救卵胞浆内单精子显微注射术(Re-ICSI)分为常规体外受精(IVF)组(472周期)和Re-ICSI组(86周期).按既往妊娠史将IVF组和Re-ICSI组分为原发不孕和继发不孕,其中IVF组原发不孕269周期,继发不孕203周期;Re-ICSI组原发不孕64周期,继发不孕22周期.IVF组和ReICSI组患者胚胎种植率、临床妊娠率、早期流产率的比较采用x2检验,活产率的比较采用Fisher精确概率检验.IVF组和Re-ICSI组间不孕年限、前向运动精子数量、精子畸形率、精子顶体酶活性的比较采用Mann-Whitney U检验.结果 IVF组胚胎种植率、临床妊娠率、早期流产率、活产率分别为29.4％、44.9％、13.4％、37.0％,Re-ICSI组分别为25.7％、34.6％、10.7％、29.6％,差异均无统计学意义(x2=0.869、2.963、0.010,P=0.351、0.085、0.922、0.098).IVF组中,原发不孕患者的不孕年限、前向运动精子数量、精子畸形率、顶体酶活性分别为4.00(3.00 ～6.00)年、58.37( 33.64 ～102.27)×106条、81.09％ (79.41％～88.69％)、76.30(48.50 ～92.46) μIU/106条精子,Re-ICSI组分别为5.00 (3.25～8.00)年、36.33 (20.59 ～ 64.43)×106条、85.50％ (81.28％ ～ 89.02％)、47.14(31.61 ～90.24) μIU/106条精子,差异均有统计学意义(Z=-2.617、- 3.505、- 3.553、-3.530,P=0.009、0.000、0.000、0.000).IVF组中,继发不孕患者的不孕年限、前向运动精子数量、精子畸形率、顶体酶活性分别为5.00(3.00～7.00)年、63.00( 34.20 ～ 107.73)×106条、81.29％(79.90％ ～ 86.09％)、78.34( 53.87 ～ 98.00)μIU/106条精子,Re-ICSI组分别为5.00 (3.75 ～7.00)年、28.80(18.57 ～48.56)×106条、88.79％(84.04％ ～95.64％)、54.70(39.73 ～76.77) μIU/106条精子,除不孕年限外,其他各项指标差异均有统计学意义(Z=-0.338、-3.505、-3.553、-3.530,P =0.735、0.000、0.000、0.006).结论 患者的不孕年限、前向运动精子数量、精子畸形率、顶体酶活性对短时受精后是否出现受精失败有预测价值.