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。     

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。     

低氧对大鼠精子体外顶体反应和顶体酶、透明质酸酶活性的影响

目的研究低氧对大鼠精子顶体酶、透明质酸酶活性的影响以及对精子体外顶体反应发生的影响。方法成年Wistar大鼠随机分为4个组:常氧对照组、低氧5d组、低氧15d组和低氧30d组。低氧组置低压舱内模拟高原5000m,分别低氧5、15和30d。采用明胶薄膜法、透明质酸底物转化法、溶血磷脂胆碱(LPC)诱导金霉素荧光染色法,测定低氧对附睾尾精子顶体酶、透明质酸酶活性和体外顶体反应发生率的影响。结果低氧5、15和30d组大鼠精子顶体反应发生率,与常氧对照组相比均显著降低,分别由常氧对照组的(30.5±3.5)%,下降至(11.7±0.9)%(P<0.01)、(10.8±1.0)%(P<0.01)和(10.0±1.4)%(P<0.01);低氧5、15和30d组大鼠精子顶体酶阳性率降低,分别由常氧对照组的(81.67±7.16)%,下降至(54.17±3.82)%(P<0.01)、(30.00±3.92)%(P<0.01)和(43.00±3.63)%(P<0.01);低氧各组透明质酸酶活性无显著改变(P>0.05)。结论低氧抑制了精子顶体酶活性和顶体反应发生。     

重组人肿瘤坏死因子-α对人正常精子运动影响的实验研究

目的 观察重组人肿瘤坏死因子-α(rhTNF-α)在体外对人正常精子运动的影响.方法 40例健康自愿者正常精液,Peroll梯度离心将精子密度调整为10×106/ml,分别与对照组以及终浓度为30 pg/ml、60 pg/ml、90 pg/ml、270 pg/ml的rhTNF-α孵育.在0.5h、1h、2h、3h、4h时间段取样,用CASA检测精子运动参数变化.结果 rhTNF-α终浓度为60、90、270 pg/ml各组精子的活力、直线速度、曲线速度、平均速度和前向运动明显低于对照组,差异均有统计学意义(P＜0.01),且270 pg/ml组的rhTNF-α对精子作用最为显著.结论 一定浓度的rhTNF-α可明显抑制人正常精子运动.     

肿瘤坏死因子受体在人精子的表达及其意义

目的　证实肿瘤坏死因子受体 (TNF R2 )在人精子有表达 ,为TNF α对精子的直接作用提供实验证据。　方法　用Western印迹法检测TNF R2 在 15例生育者精子的表达。　结果　 15例生育者的精子均有TNF R2 表达。　结论　TNF R2 在人精子有表达 ,该受体可能参与了TNF α在人精子的信号转导及对精子功能的调控     

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。     

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。     

精子诱发顶体反应与精子顶体酶活性测定关系的研究

目的 通过对精液常规正常的不育男子检测人类精子诱发顶体反应和测定精子顶体酶活性,探讨两种检测结果 的相关性.方法 40例精子密度≥20×106/ml、活动力(a+b)级精子≥50%的不育男子精液,采用离子载体A23187诱导精子顶体反应(AR),用FITC-PSA荧光染色法分析AR,并计算顶体反应发生率;用分光光度比色法(Kennedy法)测定精子顶体酶活性.用组间方差分析方法 比较精子顶体反应发生率与精子顶体酶活性之间关系.结果 两种检测结果 没有明显相关性(r=0.292,P>0.05).结论 两种检测联合应用可作为临床分析男性不育病因的检测手段.     

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。     

Roles of tumor necrosis factor alpha on sperm acrosin activity and acrosome reaction

Aim: To study the roles of tumor necrosis factor alpha (TNF-a)on the sperm acrosin activity and acrosome reaction. Methods:The sperm acrosin activity was tested by the method of BAEE/ADH Unity and the acrosome reaction by the Triple-stain technique. Results: TNF-a decreased the sperm acrosin activityand acrosome reaction (P＜0.01, P＜0.01, respectively);     

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.     

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.     

Inhibitory effects of tumor necrosis factor alpha on fracture healing in rats

Fracture healing, which involves a cascade of biological tissue responses, may be affected by various biochemical substances. One of these substances is tumor necrosis factor alpha (TNF). Studies were made on the effects of TNF on healing of fractured ribs of rats. Fracture healing was inhibited by daily administration of recombinant human TNF (400 μg/kg body weight per day, intraperitoneally) after fracture. The rate of union on day 20 was significantly lower in the TNF-treated group (4/18, 22.2%) than in the control group (14/18, 77.8%) (p < 0.001 by Chi-square test). Histological examination showed that TNF inhibited cartilagenous callus formation. On day 10, cartilage was seen in the gap zone and under the periosteum in the control group, but no cartilage formation was observed in the gap zone in 9 of 12 specimens from the TNF-treated group. On day 20, the fracture ends were united by newly formed bone in the control group, but mature fibrous tissue was seen in the gap zone, and bony or cartilagenous union was not achieved in the TNF-treated group. These results show that TNF inhibits cartilage formation in the early phase of bone induction in fracture healing and suggest that this effect of TNF is due to its inhibition of differentiation of mesenchymal cells into chondroblasts.     

Stimulation of capacitation and the acrosome reaction in ejaculated buffalo (Bubalus bubalis) sperm and the effects of a sperm motility factor

The conditions for stimulation of in vitro capacitation and the acrosome reaction of ejaculated buffalo sperm has been determined. Washed ejaculated sperm were successfully capacitated in BWW medium supplemented with bovine serum albumin (BSA) and a sperm motility factor(s) (SMF(s) isolated from the adrenal glands of rats. The acrosome reaction was induced in capacitated sperm by introducing Ca++ ions (final concentration 5 mM) into the medium. Supplementation of BWW medium with SMF(s) in the presence of BSA significantly increased the percentage of sperm showing capacitation and the acrosome reaction. SMF(s) also significantly increased the percentage motility, the percentage forward motility and maintained a higher percentage of live sperm in BWW medium under the conditions used in this study. The significance of the present findings is discussed.     

1077例男性不育患者精子顶体酶活性及其影响因素分析

本研究对１０７７ 例不育门诊病人的精子进行顶体酶活性检测,并分析了某些因素对顶体酶活性的影响。研究发现,精子顶体酶活性呈正偏态分布,６５ ．５ ％ 低于正常值（１８ μ Ｉ Ｕ／１０６ 精子） ;相关检验表明,顶体酶活性与精子活率、活力呈显著正相关,与精子畸形率和镜下精液白细胞呈显著负相关;卡方检验表明,顶体酶活性与支原体感染、精浆抗精子抗体无关;秩和检验表明,顶体酶活性降低组精液粘度高于顶体酶活性正常组（ Ｐ＜ ０ ．０００５） 。