Relationship between acrosin activity of human spermatozoa and oxidative stress

Aim: To study the association between seminal oxidative stress and human sperm acrosin activity. Methods: It is a prospective study consisting of 30 infertile men and 12 fertile normozoospermic volunteers. A full history, clinical examination and scrotal ultrasound were done to exclude other related factors such as smoking and varicocele. Presence of white blood cells (WBCs) in semen samples was evaluated by peroxidase staining. Lipid peroxidation in spermatozoa was induced after incubating with ferrous sulphate (4 mmol/L) and sodium ascorbate (20 mmol/L). Induced peroxidation of spermatozoa was assessed by determining the production of thiobarbituric acid reactive substances (TBARS). Acrosin activity was measured using the gelatinolysis technique. The halo diameters around the sperm heads and the percentages of spermatozoa showing halo formation were evaluated. An acrosin activity index was calculated by multiplying the halo diameter by the halo formation rate. Results: A significant difference was observed in acrosin activity parameters and TBARS levels between samples with WBCs (>1×106/mL of ejaculate) and those without. This difference was also noted between the normozoospermic and the oligoasthenoteratozoospermic semen samples. The TBARS production by spermatozoa had a significant negative correlation with the acrosin activity index (r = -0.89, P <0.001). Conclusion: The presence of oxidative stress in an individual with leukocytospermia and/or abnormal semen parameters is associated with impaired sperm function as measured by its acrosin activity.

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 acrosin activity.

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）

Synthesis and Crystal Structure of a Novel Ethyl 5-(4-(2-Phenylacetamido)phenyl)-1H-pyrazole-3-carboxylate as an Acrosin Inhibitor

The title compound （ethyl5-（4-（2-phenylacetamido）phenyl）-lH-pyrazole-3-carboxylate, C20H19N3O3） was synthesized by the reaction of Claisen condensation, cyclization, reduction and acylation. The structure was characterized by X-ray diffraction, MS, NMR and IR. It belongs to the monoclinic system, space group C2/c with a = 22.723（9）, b = 9.324（4）, c = 18.890（8） A, β = 114.259（6）°, V = 3649（3） A^3, Dc = 1.272 Mg·m^3, Z = 8, Mr = 349.38, p = 0.087 mm^-1, F（000） = 1472, the final R = 0.0615 and wR = 0.1643. The biological test shows that the title compound has a moderate acrosin inhibition activity.

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

Aim: To study the roles of tumor necrosis factor alpha (TNF-α) 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-α decreased the sperm acrosin activity and acrosome reaction (P<0.01; P<0.01, respectively); it also inhibited the Ca2+-ATPase activity and SOD activity in sperm (P< 0.05; P<0.001, respectively), but increased the NOS activity and the amount of NO in sperm (P<0.001; P<0.001, respectively). While it had a less significant effect on the activity of Na+-K+-ATPase (P>0.05). Conclusion: TNF-α inhibits the sperm acrosin activity and acrosome reaction.

Sperm acrosin activity may be a useful factor in choosing between ICSI and IVF for infertile male patients

The clinical applications of acrosin activity are limited.We analyzed 61578 male partners in infertile couples who visited the outpatient department of the Reproductive and Genetic Hospital of CITIC-Xiangya(Changsha,China)between August 2014 and December 2019 to determine the reference ranges and thresholds for acrosin activity in infertile Chinese men;to determine whether correlations exist between acrosin activity and age,sperm concentration,sperm morphology,or sperm motility;and to evaluate whether acrosin activity could serve as an effective prognostic indicator for choosing between in vitro fertilization(IvF)and intracytoplasmic sperm injection(icsl)in the clinic.The cut-off value for the normal reference range of acrosin activity for male partners in infertile couples was 24.78μlU per 106 sperm.There was no significant association between acrosin activity and age,sperm concentration,semen volume,total sperm count,progressive motility,or total motile spermatozoa.A weak positive correlation was found between acrosin activity and normal sperm morphology.There was a statistically significant difference in abnormal acrosome morphology between the group with high acrosin activity(>24.78μlU per 106 sperm)and the group with low acrosin activity(<24.78μlU per 106 sperm).The group with a low IVF fertilization rate had a high index of abnormal acrosomal morphology at 21.2%,while the group with a high IVF fertilization rate had a low index of 0.2%.At an acrosin activity of<24.78μlU per 10 sperm,in one cycle of the same patient,the fertilization rate,normal fertilization rate,and good-quality embryo rate for Icsl were significantly higher than those for IVF.Therefore,the most promising application of acrosin activity could be in the selection of IcsI over IVF for infertile male patients with complete fertilization failure or a low fertilization rate.

Acrosin structure-based design,synthesis and biological activities of 7-azaindol derivatives as new acrosin inhibitors

A series of 7-azaindol derivatives were designed based on the homologous 3D model of human acrosin.These compounds were synthesized and evaluated for their human acrosin inhibitory activities in vitro.Compounds 7a,7i,7j,7k and 7n showed highly inhibitory activity against human acrosin.The three-dimensional structure-activity relationship was investigated through a CoMFA model,which provided valuable information to further study of potential human acrosin inhibitors.

精子相关参数与体外受精受精率的相关性研究

目的探讨精子参数与体外受精(IVF)受精率的相关性。方法回顾性分析2020年4月至2023年4月在郴州市第一人民医院生殖医学中心进行IVF助孕的符合纳入、排除标准的327例病例。依据我中心胚胎实验室质控数据考核标准分为低受精组(受精率<30%,n=19)和正常受精组(受精率≥30%,n=308)。收集男方的基本信息与精子相关参数以及胚胎信息。用Pearson相关性分析各参数与精子受精率的相关性,Logistic回归分析精子受精率的影响因素,采用ROC曲线分析预测价值。结果精子顶体酶活性、正常形态精子百分率、精子DNA碎片指数(DFI)是影响受精率的相关因素,相关系数(r)分别为0.168、0.306、-0.243(P均<0.05)。Logistic回归分析确定三者均是影响IVF受精率的独立因素,并得出多因素回归方程式:受精率预估值=-2.561+0.035×精子顶体酶活性+2.066×正常形态精子百分率-1.51×精子DFI。精子顶体酶活性、正常形态精子百分率、精子DFI及三因素联合预测低受精率的ROC曲线下面积(AUC^(ROC))分别为0.806、0.889、0.827、0.899,其Yuden指数对应的cut-off值分别为61.2μIU/10^(6)精子、2.98%、27.50%、-21.32,敏感性分别为95.70%、76.90%、68.40%、64.30%,特异性分别为43.50%、94.70%、87.70%、100.00%,表明3个指标联合预测受精率的效能较好。结论精子顶体酶活性、正常形态精子百分率、精子DFI是预测IVF受精率的独立影响因素;可以用多因素回归方程式预测IVF的受精率。

上皮性卵巢癌症患者血清和肿瘤组织中ACRBP表达与一线化疗效果和预后的关系

目的探讨上皮性卵巢癌(EOC)患者血清和肿瘤组织中顶体素结合蛋白(ACRBP)表达与一线化疗效果和预后的关系。方法使用从基因表达谱互动分析数据库中获得的数据分析卵巢癌(OC)中ACRBP mRNA的表达和预后作用。收集2019年10月至2021年12月在该院接受手术的95例EOC患者的组织标本和血清样本。另外,收集30例卵巢良性肿瘤物组织样本作为良性肿瘤对照组,收集30例健康志愿者血清样本作为健康对照组。采用免疫组织化学染色法检测组织ACRBP表达。一线化疗完成后,使用实体肿瘤疗效评价标准1.1版评估化疗效果分为敏感组和耐药组。敏感组包括完全缓解(CR)者和部分缓解(PR)者,耐药组包括疾病稳定(SD)者和疾病进展(PD)者。结果公共数据库中,OC患者组织ACRBP mRNA表达明显升高(P<0.05),且其高表达与总生存期(OS)较差有关。临床样本中,ACRBP主要在EOC肿瘤细胞的细胞质中表达,而在良性对照组卵巢组织中均未检测到ACRBP。一线化疗耐药组患者组织ACRBP表达显著高于敏感组,差异有统计学意义(P<0.001)。组织ACRBP表达预测一线化疗耐药的受试者工作特征(ROC)曲线下面积(AUC)为0.830(95%CI:0.743~0.916),此时的截断值为8.65分,灵敏度和特异度分别为80.0%和77.1%。ACRBP高表达是EOC患者OS和PFS预后的独立危险因素(P<0.05)。根据组织ACRBP表达中位值,将EOC患者分为ACRBP低表达组(<8.27分)和高表达组(≥8.27分)。组织ACRBP高表达组患者OS和无进展生存期(PFS)明显更短(P<0.05)。结论EOC患者组织ACRBP高表达与一线化疗耐药和预后不良密切相关。ACRBP有希望成为一线化疗效果和预后预测的新型生物标志物。

精子顶体酶活性与精液常规参数的相关性

目的 :测定人精子顶体酶活性并了解该酶与精液常规分析有关参数之间的关系 ,寻求一种可靠的衡量精子生育力的参数。方法 :分光光度比色法测定精子顶体酶活性。结果 :顶体酶活性与精子密度、a及 b级活动力精子百分率、精子活动率之间存在明显正相关 ( P<0 .0 0 1 ) ,而与畸形精子率呈显著负相关 ( P<0 .0 0 5 )。以精液常规分析各主要参数正常与否分成的两组间其顶体酶活性差异也均有显著性 ( P<0 .0 0 1 )。结论

精子顶体酶检测对不明原因不孕夫妇助孕治疗方案选择的临床意义

目的:探讨精子顶体酶检测对不明原因不孕(UI)夫妇选择助孕治疗方案的临床意义。方法:回顾性分析2013年1月至2015年12月诊断为UI,并经3次宫腔内人工授精(IUI)治疗失败改行体外受精-胚胎移植(IVF-ET)助孕的49对夫妇共49个周期的临床资料,并对比分析同期因输卵管阻塞因素行常规IVF治疗的95对夫妇的131个周期的临床资料,比较两组实验室数据、临床结局和顶体酶活性差异;进一步根据UI夫妇男方精子顶体酶活性将UI组分成2个亚组:<36 IU/106精子组共20个周期,≥36 IU/106精子组共29个周期,比较两亚组之间受精率的差异。结果:1行IVF-ET治疗的UI夫妇,比同期因输卵管阻塞行常规IVF的夫妇受精率显著下降(67.0%vs 76.4%,P<0.05),补救性卵细胞胞质内单精子注射(ICSI)率高于输卵管因素组(20.4%vs 6.1%,P<0.05);成熟卵母细胞比例(MII卵率)、可利用胚胎率、优胚率、种植率、临床妊娠率两组差异无统计学意义(P>0.05)。2UI组中位精子顶体酶活性低于输卵管因素组(36.03 IU/106精子vs 61.98 IU/106精子,P<0.01),UI组中顶体酶活性<36 IU/106精子亚组的受精率明显低于顶体酶活性≥36 IU/106精子亚组的受精率(47.7%vs80.3%,P<0.01)。结论:1UI夫妇男方精子顶体酶活性低下导致的受精率低,可能是造成女性不孕的主要原因和潜在因素。2鉴于UI夫妇男方精子顶体酶活性<36 IU/106精子时受精率较低,建议不采用IUI治疗,可选择IVF+短时受精联合早期补救ICSI治疗。

左卡尼汀对精子顶体酶活性低下男性不育患者的疗效分析

目的:观察左卡尼汀(LC)治疗精子顶体酶活性低下男性不育患者的疗效。方法:收集精子顶体酶活性低于正常值的男性不育患者212例,随机分为两组,治疗组口服LC 1. 0 g/次,3次/d;对照组给予维生素E软胶囊100 mg/次,3次/d,连续用药3个月。治疗前后分别检测精液常规参数与精子顶体酶活性。根据精液常规分析结果,将治疗组进一步分为少精子症组、弱精子症组和正常组。结果:治疗组中,与治疗前相比,治疗后前向运动精子百分率(PR%)明显提高[(36. 35±1. 26)%vs (32. 58±1. 13)%,P <0. 05],精子顶体酶活性显著提高[(58. 61±1. 93)μIU/106精子vs (37. 05±0. 66)μIU/106精子,P <0. 01]。而在对照组中,治疗后精子浓度、PR%、顶体酶活性较治疗前都有提高,但无显著差异。将治疗组进一步分组分析,治疗后少精子症组精子浓度较治疗前显著增高[(21. 82±4. 21)×106/ml vs (11. 27±0. 73)×10~6/ml,P <0. 01],弱精子症组PR%较治疗前显著提高[(29. 81±1. 88)%vs (20. 61±0. 85)%,P <0. 01],精子顶体酶活性弱精子症组较正常组和少精子症组提高更加显著[(60. 85±3. 04)μIU/106精子vs (56. 32±2. 86)、(57. 09±6. 31)μIU/106精子,P <0. 05]。结论:口服LC可以有效改善精子的顶体酶活性,且对弱精子症患者效果更明显。

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。

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。

精子DNA碎片率及顶体酶活性对男性不育的诊断价值

【目的】探讨精子DNA碎片率与顶体酶活性检测对男性不育的诊断价值。【方法】回顾性分析近三年前来本院就诊的902例男性患者的精液常规参数及精子DNA碎片率和顶体酶活性水平。【结果】按DNA碎片率水平将所有患者分为4组(≤10%,11~20%,21~30%,≥31%),各组患者的年龄、精子浓度、精子总活力、前向运动率以及顶体酶活性存在统计学差异;按精子顶体酶活性正常与否将患者分为两组后,两组间精子浓度、精子总数、精子总活力、前向运动率、正常形态率、畸形精子指数、DNA碎片率均有统计学差异。相关性分析显示,DNA碎片率与精子浓度、精子活力、前向运动率、精子总数及顶体酶活性呈负相关关系,与畸形精子指数呈正相关关系;顶体酶与精子浓度、精子活力、前向运动率、精子总数、正常形态率呈正相关关系,与头部异常率、畸形精子指数、DNA碎片率负相关。【结论】精子DNA碎片率及顶体酶活性水平均可以在一定程度上反映精子参数水平,其中DFI更能反映精子活力水平,顶体酶活性则与精子形态更为相关。两者都是精子质量评估更为客观而全面的指标。

精子形态与顶体酶活性关系分析

目的探讨精子形态与精子顶体酶活性的关系。方法对268例男性不育患者采用改良Kennedy法测定顶体酶活性,采用改良巴氏染色法分析精子形态,以正常形态精子率大于或等于15%为正常组,正常形态精子率小于15%为异常组。结果正常组顶体酶活性高于异常组顶体酶活性,差异无统计学意义(P=0.05);正常组正常形态精子率及头部畸形率与顶体酶活性不相关(P>0.05);异常组正常形态精子率与顶体酶活性存在正相关(r=0.39,P<0.01),头部畸形率与顶体酶活性存在负相关(r=-0.352,P<0.01)。结论精子形态可影响顶体酶活性。

男性不育患者精子形态与精浆锌和精子顶体酶活性关系的研究

目的：研究男性不育患者精子形态与精浆锌和精子顶体酶活性关系。方法455例男性不育患者分别根据年龄和精子正常形态百分率分成5组和8组，采用 Diff-Quik 染色法、改良 PRA 法和改良 Kennedy 法分别进行精子形态、精浆锌和精子顶体酶活性的检测。结果各年龄组的正常形态精子百分率和精浆锌浓度差异无统计学意义（P ＞0．05），精子顶体酶活性在36～50岁年龄段显著降低（P ＜0．05），其他年龄段精子顶体酶活性差异无统计学意义（P ＞0．05）。在精子正常形态百分率不同的8组中，年龄差异无统计学意义，精浆锌总量差异无统计学意义，精子顶体酶活性随着精子正常形态百分率降低而降低（r ＝0．93，P ＜0．01）。结论精子正常形态百分率不受年龄和精浆锌的影响，与精子顶体酶活性呈正相关。

顶体蛋白酶抑制剂4-胍基苯甲酸盐对人精子功能的影响

目的 :研究顶体蛋白酶抑制剂 4-胍基苯甲酸盐 (KF- 95 0 )对人精子功能的影响。方法 :体外用不同浓度的 KF- 95 0作用于人精子 ,观察其活动率 ;用免疫组化 ABC法观察 KF- 95 0对精子顶体结构的影响。结果 :KF- 95 0对人精子的作用与其浓度和作用时间有关。经≥ 0 .2 mg/ ml KF- 95 0处理 ,随着时间的延长以精子活动率明显下降 ,精子膜功能有不同程度的损害 ,顶体脱落率以及已发生顶体反应的精子死亡率明显上升。 结论 :KF- 95

解脲支原体感染对不育男性精液质量的影响

目的:研究解脲支原体(UU)感染对不育男性精液质量的影响.方法:收集不育男性精液143例,分为UU阳性的观察组46例和UU阴性的对照组97例;采用西班牙CASA计算机辅助分析系统分析精子活力,通过伊红染色法检测精子存活率,精子形态染色则采用标准巴氏法,精子顶体酶活性、表面结合抗体As Ab水平、精浆弹性蛋白酶水平均采用试剂盒进行检测.结果:与对照组相比,UU感染组精子正常形态率显著降低(P<0.05),而顶体酶异常率显著升高(P<0.05);精子前向运动率、总活力、存活率及顶体酶活性降低,但无统计学差异(P>0.05);精子畸形指数、畸形精子指数、As Ab水平及精浆弹性蛋白酶水平均无统计学差异(P>0.05).结论:UU感染可导致精子正常形态率显著降低,顶体酶异常率显著升高,从而使男性不孕率升高;同时,UU感染亦可能影响精子活动率、存活率、顶体酶活性、As Ab水平及精浆弹性蛋白酶水平.

精子顶体酶活性与精液有关参数的研究

目的分析精子顶体酶活性与精液有关参数变化,探讨精子顶体酶活性对男性不育的影响。方法对214例精液标本作精子顶体酶活性、弹性硬蛋白酶、果糖、α-葡萄糖苷酶、锌、酸性磷酸酶、精子尾部低渗肿胀试验检测,同时用WLJY-9000伟力彩色精子质量检测仪作精液分析。以精子顶体酶活性正常的精液作为对照组,以精子顶体酶活性异常的精液作为观察组,并对两组参数进行比较。结果214份精液标本,精子顶体酶活性正常111例,异常103例。对照组在精子密度、精子活动率、a+b级精子百分率、精子尾部低渗肿胀试验均明显优于观察组(P<0.001),弹性硬蛋白酶、果糖、α-葡萄糖苷酶均明显优于观察组(P<0.05),精液量、锌、酸性磷酸酶无差别(P>0.05)。结论精子顶体酶活性与精液有关参数关系密切,精子顶体酶活性降低将能引起男性不育。