低氧对大鼠睾丸生殖细胞凋亡的影响

目的:研究低氧对大鼠睾丸生殖细胞凋亡和Bax、Bcl-2表达的影响。方法:雄性成年Wistar大鼠随机分为4组:常氧对照组、低氧5 d组、低氧15 d组和低氧30 d组(各组n=6)。常氧对照组在平原喂养;低氧5 d组、15 d组和30 d组分别在低压舱内模拟5 000 m高原喂养5、15、30 d。采用流式细胞术和TUNEL法检测低氧对睾丸生殖细胞凋亡的影响。运用Western印迹技术检测低氧对大鼠睾丸内凋亡相关蛋白Bax、Bcl-2表达的影响。结果:低氧5 d组、15 d组和30 d组睾丸内发生生殖细胞凋亡的生精小管数量[每100个生精小管中,含有凋亡生殖细胞的生精小管数分别为(20.50±5.07)、(21.25±7.85)、(14.00±2.45)个]均非常显著地高于常氧对照组[(6.00±2.16)个,P<0.01]。凋亡的生殖细胞以精原和精母细胞为主。低氧15 d组睾丸组织亚单倍体细胞百分率[(2.18±0.82)%]显著高于常氧对照组[(1.30±0.33)%,P<0.05],低氧30 d组[(3.08±0.93)%]极显著高于常氧对照组(P<0.01)。低氧30 d组睾丸组织Bax表达显著高于常氧对照组(P<0.05),其灰度值分别为17.34±4.54和10.50±2.82。低氧30 d组睾丸组织Bax/Bcl-2比值显著高于常氧对照组(P<0.01),比值分别为0.40±0.10和0.27±0.04。结论:低氧诱导大鼠睾丸生殖细胞凋亡增多,慢性低氧引起的睾丸生殖细胞凋亡增多与Bax表达增加有关。     

低氧对大鼠附睾功能的影响

目的研究低氧对大鼠附睾功能的影响。方法将成年雄性Wistar大鼠随机分为常氧和低氧5d、15d、30d4组。低氧各组置低压舱内模拟5,000m高原低氧。测定附睾尾精子数量、质量,生化法测定各组大鼠附睾液中果糖、肉毒碱的含量及α-葡萄糖苷酶的活性。结果低氧15d组、30d组大鼠附睾尾精子数显著低于常氧组。低氧各组精子活力、活率显著低于常氧组;尾部畸形精子数显著高于常氧组。低氧5d组、15d组大鼠附睾液果糖含量显著高于常氧组(P<0.01)。低氧5d、30d组,大鼠附睾液中性α-葡萄糖苷酶活性显著低于常氧组。结论低氧可抑制附睾尾精子数量和质量,并抑制附睾液中性α-葡萄糖苷酶活性,提示低氧可干扰附睾功能,并抑制精子的成熟。     

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。     

精子DNA损伤、核蛋白组型转换与顶体酶活性及精液参数的相关性分析

目的:探讨精子DNA损伤、精子核蛋白组型转换与顶体酶活性及精液参数的相关性。方法:收集535例精液标本,采用精子染色质扩散(sperm chromatin dispersion,SCD)检测精子DNA损伤,并与精子核蛋白组型转换、顶体酶活性和WHO手册(第4版)精液参数进行相关性分析。结果:精子DNA损伤与精子核蛋白组型转换、顶体酶活性、精子浓度及前向运动精子这些指标之间比较均具有显著性差异(P<0.01);精子DNA损伤与年龄、核蛋白组型转换、精子浓度和D级精子比例之间呈显著正相关(P<0.01或P<0.05),而与顶体酶活性呈显著负相关(P<0.001),多元逐步回归分析显示年龄、精子浓度、D级精子比例、核蛋白组型转换及顶体酶活性是5个独立的相关变量。精子核蛋白组型转换、顶体酶活性、精子密度和前向运动精子这4个指标的异常率在精子DNA异常组(DFI≥30%)中均显著的高于正常组(DFI<30%)。结论:精子DNA损伤与精子核蛋白组型转换、顶体酶活性及WHO(第4版)精液各参数之间存在密切的联系,可能是评价精子质量的另一项重要的指标。     

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.     

217例男性不育症精子顶体酶检测与分析

目的　研究人精子顶体酶与男性不育之间的关系。　方法　 2 1 7例不育男性 ,年龄 2 4～ 43岁 ,根据精液常规检查结果分为不育 A、B两组 ;46例生育男性为对照组。手淫留取精液标本 ,常规洗涤后孵育 ,采用固定明胶底膜法检测精子顶体酶。　结果　正常生育组的精子顶体酶活性阳性率为 86.5% ,酶活性亮区直径为 42 .9μm。不育 A组酶活性阳性率为 57.0 % ,酶活性亮区直径为 3 8.5μm。不育 B组酶活性阳性率为3 5.2 % ,酶活性亮区直径为 2 7.5μm。经统计学处理 ,生育组与不育组之间精子顶体酶阳性率差异有显著性 ( P<0 .0 1 )。　结论　顶体酶阳性率和活力低下是男性不育的一个重要原因 ,精子顶体酶是受精过程中的关键酶     

精浆抗精子抗体阳性不育患者顶体酶、一氧化氮合酶及超氧化物歧化酶活力变化的研究

目的:研究精浆抗精子抗体(AsAb)阳性对精子顶体酶、精浆一氧化氮合酶(NOS)及超氧化物歧化酶(SOD)活力的影响。方法:精浆AsAb阳性不育者40例,对照组为40例正常生育男性。通过吸光度变化分别计算顶体酶活力(BAEE/ADH联合法)、NOS活力(氧化还原反应)、SOD活力(黄嘌呤氧化酶法)。结果:精浆As-Ab阳性组与正常生育组比较,精子顶体酶活力明显降低(P<0.01),NOS活力明显升高(P<0.01),精浆中SOD活力明显降低(P<0.01)。结论:精浆AsAb阳性引起不育可能与精子顶体酶、精浆中SOD及NOS活力改变有关。     

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。     

顶体酶活性对体外受精-胚胎移植结局的影响

目的:探讨精子顶体酶活性对体外受精-胚胎移植(IVF-ET)结局的影响。方法:选择909对在本院生殖中心就诊的不孕夫妇,其中女方检查为正常或仅为输卵管因素不孕。采用改良巴氏法染色和Na-苯甲酰-DL-精胺酸-P-硝酰基苯胺(BAPNA)法分别对丈夫的精液进行精子形态学分析和精子顶体酶活性测定。结果:顶体酶异常组中各项精液参数,包括正常形态精子百分率、精子活动率、活动力、快速前向运动精子百分率及精子密度均显著低于顶体酶正常组(P<0.01);顶体酶活性与精液常规分析上述各参数之间存在非常显著的正相关(P<0.01);两组间取卵数、卵裂率、优质胚胎率、胚胎冷冻率、无可移植胚胎周期百分率、胚胎移植数和流产率均无显著性差异(P>0.05);顶体酶正常组的受精率、仅一个胚胎移植的周期百分率、着床率及临床妊娠率均明显高于顶体酶异常组(P<0.01)。结论:顶体酶活性异常与精液常规分析各主要参数的异常密切相关,可导致体外受精率显著下降。顶体酶活性检测在IVF结局预测中起着重要作用。     

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     

人精子顶体反应的检测及其与穿卵试验的相关性

选择正常生育男子（生育组）、输精管结扎再吻合（吻合组）、不育症患者（不育组）各２５例的精液进行人精子顶体蛋白酶及酸性磷酸酶测定,顶体组织化学三色染色和人精子穿透去透明带金黄仓鼠卵试验,比较不同生育条件下精子功能和生育能力的相关性。结果显示,精子获能后顶体反应率明显高于获能前（Ｐ＜０．０１）。生育组顶体反应率和穿透试验高于吻合组和不育组（Ｐ＜０．０１）。结果表明,精子穿卵试验结合精子顶体反应测定可作为客观评价人类精子受精能力的依据     

精子功能指标在辅助生殖助孕方式选择中的作用

目的 评估精子功能参数对精子常规体外受精能力的预测价值,探寻有效选择受精方式的新指标.方法 回顾性分析2017年1月至2019年12月于我院首次行体外受精-胚胎移植(IVF-ET)短时受精的429个周期的临床资料,根据受精结局不同分为2组:受精失败或受精率＜30％的患者为早补救卵胞浆内单精子注射(R-ICSI)组(n=...     

Correlation between sperm morphology, acrosin, and fertilization in an IVF program

Acrosin, a neutral proteinase, is located within the acrosome. The aim of this study was to evaluate acrosin concentrations in patients with severe damage of the sperm head and to determine whether acrosin concentration could predict the chances of fertilization in an IVF program. Sixty patients were accepted into this study, prospectively. The patients were divided into two groups, those with a normal morphology of less than 14% (group I, n = 33) and those with normal morphology less than 14% (group II, n = 27). All patients had a sperm concentration of less than 20 million sperm/ml and less than 30% progressively motile sperm. The acrosin assays were performed on the semen sample obtained on the day of IVF. Routine IVF insemination procedures were used, and only mature oocytes were considered. The only factor that showed a significant correlation of fertilization was normal morphology (p less that 0.01). The mean acrosin level was 73.4 /+- 38.6 mED/10 million sperm in group I and 70.9 /+- 42.7 mIU/10 million sperm in group II (no significant difference). The fertilization rate in group I was 45.4% and in group II, 77.7% p less than 0.002). Acrosin levels were not significantly different in patients with and without fertilization (72.0 /+- 42.1 and 73.6 /+- mIU/10 million sperm, respectively).     

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.     

干扰素-γ和肿瘤坏死因子-α对精子受精能力的影响及机制初探

目的:观察IFN-γ和TNF-α对正常精子顶体酶活性和顶体反应率的影响,并对其变化机制进行初步探讨。方法:36例精液常规分析基本正常标本,经75%Percoll分离后,分别或联合使用IFN-γ和TNF-α处理(终浓度均为30ng/ml)。采用BAEE/ADH联合法测定精子顶体酶活力的变化。用三色染色法观察精子顶体反应率的变化。用高效液相色谱法(HPLC)检测精子NO含量。用试剂盒法测定Na+-K+-ATPase、Ca2+-ATPase和SOD活性。结果:IFN-γ和TNF-α单独及联合使用时,均可显著降低精子顶体酶活性和顶体反应率(P<0.05或P<0.01),且TNF-α的抑制作用更强些;IFN-γ可使精子Na+-K+-ATPase、Ca2+-ATPase和SOD活性显著降低(P<0.01),且两者协同作用更低,但TNF-α对Na+-K+-ATPase和Ca2+-ATPase活性几乎无作用;IFN-γ、TNF-α及两者合用时均使精子NO含量显著增加(P<0.01)。结论:IFN-γ和TNF-α对精子顶体酶活性及顶体反应率有一定的抑制作用,并且可能通过对精子Na+-K+-ATPase、Ca2+-ATPase、SOD活性以及NO含量等多方面影响来实现的。     

IgA抗精子抗体对精子顶体反应的影响

目的 探讨精浆中IgA抗精子抗体对人精子顶体反应的影响。方法 利用免疫珠法（IBT）筛选出IgA抗精子抗体阳性精浆标本同正常人精子孵育，以孕酮诱发精子顶体反应；以特异性荧光标记物．络合异硫氰酸荧光素的花生凝集素（FITC-PNA）标记精子顶体，通过流式细胞仪检测精子顶体完整性。结果 与IgA抗精子抗体阳性精浆孵育的精子，其孕酮诱发的顶体反应发生率明显低于正常精浆及精子培养液组（P〈0．01），正常精浆组及精子培养液组间无显著性差异（P〉0．05）；IgA抗精子抗体阳性精浆组、正常精浆组、精子培养液组自发顶体反应的发生率无显著性差异（P〉0．05）。结论 免疫性不育患者精浆中的IgA抗精子抗体可以明显抑制孕酮诱发的顶体反应的发生，可能是导致不育的原因之一。     

Evaluation of the acrosome reaction and viability in buffalo spermatozoa using two staining methods: the effects of heparin and calcium ionophore A23187

In this study, we investigated the effect of heparin and calcium ionophore A23187 on in vitro induction of buffalo sperm acrosome reaction (AR). Two methods for detection of the AR and viability were employed. Fluorescein isothiocyanate-conjugated Arachis hypogea agglutinin (FITC-PNA) was used as a vital stain in combination with ethidium homodimer-1 (EthD-1) to determine the acrosome status of viable spermatozoa. In another experiment, trypan blue replaced EthD-1 to differentiate live and dead spermatozoa having undergone AR. The results from the two methods were significantly correlated (r > 0.9). Four different staining patterns were found in both methods. The FITC-PNA intensely labels the acrosome region of acrosome-intact spermatozoa. EthD-1 and trypan blue stained red and blue at the post-acrosomal region of dead spermatozoa, respectively. Spermatozoa incubated with heparin showed a significant increase ( p < 0.05) in the percentage of live acrosome-reacted sperm after 30 min incubation period. This trend continued and was significantly different over the entire incubation period when compared with the control group at the same interval. In the ionophore-treated group, the proportion of changes in live acrosome-intact and live acrosome-reacted spermatozoa was statistically significantly different ( p < 0.001) when compared with those treated with heparin at the same interval. The AR occurred sooner and to a greater extent when incubated with the ionophore but at 5 h of incubation the percentage of false acrosomal reaction was significantly higher than those in the control and heparin-treated groups. The results in this study indicated that the in vitro induction of AR by heparin and calcium ionophore evaluated by both methods could be used to assess sperm fertilizing capacity for in vitro fertilization of this species.