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.

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）

Studies on the Relationship between Urokinase Plasminogen Activator(uPA)and Human Sperm Motility

To clarify the role of urokinase plasminogen activator(uPA) in the mechanisms of regulating sperm motility and the ability of fertilizing, we investigated the quantities and activities of uPA in human seminal Plasma and on the membrane of spermatozoa.Semens were harvested from 22 infertile patients with asthenospermia and 20 healthy fertile men according to WHO standards. To quantify the membrane-bound uPA in the samples, polyclonal antibodies against human urokinase were employed by means of a sandwich ELISA. The uPA activities in seminal plasma and on the surface of spermatozoa were determined using Agarose-Fibrine-Plate method and the experiment of immunological identification with polyclonal antibodies against urokinase. In lysates of spermatozoa, significantly lower levels of uPA(23. 1±7.35 mu/106 cells ) and uPA activity (5.13±3.85 mu/106 cells) were found in patient group as compared to healthy fertile men exhibiting normospermia (29. 89±9. 40 mu/105 cells and 10. 17±6. 18 mu/106 cells). In seminal plasma, uPA activity in patient group (2134±1581. 3 IU/L)was also found significantly lower than that of normal group (3365±1859. 5 IU/L). Positive correlations were observed between sperm motility and uPA quantities (r=0. 48, P＜0. 005), as well as with uPA activities (r= 0. 45,P＜0. 005).Thus, it is inferred that membrane associated uPA on human spermatozoa may be related directly to sperm motility and fertility.

Objective non-intrusive markers of sperm production ant sexual activity

Objective studies of men＇s reproductive function are hindered by their reliance on： （i） self-reporting to quantify sexual activity and （ii） masturbation to quantify sperm output rendering both types of estimate vulnerable to unverifiable subjective factors. We therefore examined whether detection of spermatozoa and measurement of prostate-specific antigen （PSA） in urine could provide objective semiquantitative estimates of sperm output and recent ejaculation, respectively, using widely available laboratory techniques. Of 11 healthy volunteers who provided urine samples before and at intervals for 5 days after ejaculation, sperm was present in 2111 men before, and in all 11111 samples immediately after ejaculation, but by the second and subsequent void, spermatozoa were present in -10%. PSA was detectable at high levels in all urine samples, peaking at the first post-ejaculatory sample but returning to baseline levels by the second post-ejaculatory void. We conclude that urinary spermatozoa and PSA are objective biomarkers for sperm production and sexual activity, but only for a short-time window until the first post-ejaculatory urine void. Hence, for a single urine specimen, the presence of spermatozoa and PSA are valid biomarkers, reflecting sperm production and recent ejaculation only until the next micturition, so their measurement should be restricted to the first morning urine void.

Role of Urokinase-type Plasminogen Activator in the Precontact Sperm-egg Communication and Fertility of Mice in vitro

Objective To explore the role of urokinase-type plasminogen activator（uPA） in precontact sperm-egg communication and fertility of mice in vitro. Methods Firstly, sperm chemotaxis （SC） induced by uPA was assayed by measuring the sperm densities in capillaries with a descending gradient or no gradient of uPA respectively. Secondly, the role of uPAR that exists in sperm plasma membrane in SC was studied by examining the change of sperm density in capillary after incubating spermatozoa with anti-uPAR antibody. Thirdly, SC induced by eggs, which had been treated with uPA, PAl-1 and anti-uPAR beforehand respectively, was assayed to study the role of uPA in PSEC. Lastly, the fertilization capability of spermatozoa treated with uPA was examined by counting the number of fertilized eggs. Results 1）The density of spermatozoa that migrated down the gradient of uPA into the capillary was significantly lower than that into the capillary containing no-gradient uPA. 2） When uPAR of spermatozoa was inhibited by anti-uPAR antibody, the density of spermatozoa that migrated into the capillary with ascending gradient of uPA decreased correspondingly. 3） The density of spermatozoa attracted by eggs, which were treated with uPA beforehand, increased significantly than that of attracted by non-treated eggs. On the contrary, the sperm density decreased correspondingly when the egg was treated with PAI-1. 4） The number of fertilized eggs increased significantly after the spermatozoa used here was treated with uPA beforehand. Conclusion uPA could induce SC of mice sperm in vitro through the uPAR on its membrane, enhance the capability of egg inducing SC, and promote spermatozoa to fertilize eggs. Thus, uPA may act as an attractant in PSEC, increase the chance encounter of spermatozoa and eggs, therefore, enhance the fertility success correspondingly. This study, in some degree, provides an evidence that uPA may be used as a new medicine and diagnostic reagent for male infertility.

Studies of Ce(Ⅲ)-ALC-F Interacting with Herring Sperm DNA by Electrochemical, Fluorimetric and UV-spectrophotometric Method

Ce(Ⅲ)-ALC-F complex can react with hsDNA to form an electrochemically non-active supermolecular complex Ce(Ⅲ)-ALC-F-DNA in the buffer solution of (CH2)6N4(pH=4.9), which results in the decrease of the peak current of Ce(Ⅲ)-ALC-F. This method can be applied to determine DNA concentration. In addition, by using fluorimetric and UV-spectrophotometric methods with studies of denatured DNA and the effect of NaCl solution , it is also found that the binding mode is intercalation.

Role of platelet-activating factor in reproduction:sperm function

Since its discovery nearly thirty years ago, platelet-activating factor has emerged as one of the more important lipidmediators known. Platelet-activating factor (PAF; 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphorylcholine) exists en-dogenously as a mixture of molecular species with structural variants of the alkyl moiety. PAF is a novel potent signal-ing phospholipid that has unique pleiotropic biological properties in addition to platelet activation. PAF also plays a sig-nificant role in reproduction. PAF content in squirrel monkey sperm is significantly higher during the breeding seasonthan the non-breeding season. PAF content in human sperm has a positive correlation with seminal parameters and preg-nancy outcomes. High-fertility boars have significantly more PAF in their sperm than low-fertility boars. The enzymes(lyso-PAF-acetyltransferase and PAF-acetylhydrolase) necessary for PAF activation and deactivation are present insperm. PAF-acetylhydrolase may act as a 'decapacitation factor'. Removal of this enzyme during capacitation maypromote PAF synthesis increasing motility and fertilization. PAF also plays a significant role in the fertilization process,enhancing the fertilization rates of oocytes. Enhanced embryo development has also been reported in oocytes fertilizedwith PAF-treated sperm. PAF antagonists inhibit sperm motility, acrosome reaction, and fertilization, thus suggestingthe presence of receptors for PAF. The PAF-receptor is present on sperm, with altered transcript levels and distributionpatterns on abnormal cells. Whereas the exact mechanism of PAF in sperm function and reproduction is uncertain, itsimportance in normal fertility is substantial. The reproductive significance of PAF activity in sperm and fertility plus therole of PAF in the establishment of pregnancy requires further study.

PIWI/piRNA Found to Play a Critical Role in Translation Activation during Sperm Formation

During their formation,mouse sperms suspend their intracellular production until better timing.How do they manage to reactivate this process in due time?Now a joint team unravels this long-standing mystery.

回族及维吾尔族圆头精子症患者的基因诊断、精子超微结构观察和辅助生殖结局分析

目的探讨2例少数民族(P1回族、P2维吾尔族)圆头精子症患者的临床表型、精子特点、遗传学病因以及辅助生殖结局。方法分析2例少数民族圆头精子症患者的临床资料和各种精液检查参数,观察精子超微结构,并利用全外显子检测技术和qPCR检测分析患者的遗传学病因,采用卵胞质内单精子注射联合卵母细胞激活技术(ICSI+AOA)进行辅助生殖治疗,并观察其助孕结局。结果2例患者均存在DPY19L2基因109681 bp纯合缺失,其中P2患者的DPY19L2基因缺失来源于近亲结婚的父母。P1患者精子活力低下,精子DNA碎片率高,精子形态为100%圆头精子,电镜下发现精子顶体缺失,同时存在质膜、线粒体和微管等超微结构缺陷;P2患者精子活力及精子DNA碎片率均正常,精子形态为100%圆头精子,电镜下观察发现精子主要缺陷为头部小而圆伴随顶体缺失,质膜、线粒体和微管等细胞器结构损伤与超微结构缺陷少见。2例患者夫妇均接受ICSI+AOA助孕,ICSI受精率P1患者夫妇为62.5%,P2患者夫妇为75%,均成功获得临床妊娠。结论DPY19L2基因异常在不同民族背景的圆头精子症患者中都是主要的致病遗传学原因。圆头精子可同时存在质膜、线粒体和微管等细胞器结构损伤与超微结构缺陷。ICSI+AOA是圆头精子症患者的有效辅助生殖治疗。

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。

人类微量精子冷冻载体的研究进展

全球不孕不育症中男性因素约占50%,其中无精子症患者通常需要通过手术获取微量精子,或极重度少弱精子症患者的精子数量稀少,采用常规冷冻载体冷冻复苏后难以寻找到存活的精子。因此为避免取卵日无精子可用而取消周期,临床上应用微量精子冷冻载体预先冷冻少量精子,如空卵透明带、海藻酸胶囊、微滴法、麦管法、薄片法、剥卵针等。本文就人类微量精子的不同冷冻载体的研究进行综述,为临床上优化微量精子的冷冻方法提供理论依据。

中医药辨治免疫性不育研究概述

免疫性不育以肾虚为本、湿瘀为标,基本病机在于禀赋不足、戕害肾精、饮食不节等导致肾肝脾虚,感染、环境污染及接触化学毒物导致湿、瘀、热、毒等邪气蕴结精室,日久损伤机体免疫屏障,导致精子被机体免疫系统识别而形成抗精子抗体,引起精子活动率下降,造成不育。总结近20年本病论治概况,认为诸医家辨证运用中药、中西结合、针刺、穴位埋线等手段,整体上重视补肾治本、兼顾肝脾,活血祛湿治标、兼顾热浊,发挥了抗炎、调节免疫、提高精子活动率的作用,凸显了中医优势。

哺乳动物精子磷脂酶C zeta(PLCζ)在受精中作用的研究进展

哺乳动物受精是高度分化的雌雄配子结合,形成一个合子的过程。精卵细胞融合过程中,精子将携带的遗传物质和功能性分子释放到卵子内,引起一系列生物学事件,激活停留在减数分裂前期的卵子。精子释放的分子诱导卵子内质网中的钙离子(Ca^(2+))以一定的频率释放到胞质中形成钙振荡,作用于下游分子,打破细胞周期抑制网络,引发皮质颗粒释放,成为卵子激活的首要标志。文章综述了精子内诱发卵子激活的磷脂酶C zeta(PLCζ)和类似分子的鉴定、分子结构和作用机制方面的研究进展,并讨论了相关精子缺陷引起的人和动物的生殖障碍,以及人工卵子激活方法研究进展,以期为增进人类生殖健康和提高动物体外胚胎生产效率提供参考。

多不饱和脂肪酸对Δ15 Des转基因小鼠睾丸结构和功能的影响及机制

目的探讨多不饱和脂肪酸(PUFAs)对Δ15脂肪酸去饱和酶(Δ15 Des)转基因小鼠睾丸结构和功能的影响及机制。方法取C57BL/6野生型(WT)雄性小鼠和Δ15 Des转基因雄性小鼠分别作为对照组(n=5,WT组)和实验组(n=5,TG组),均饲喂含6%花生四烯酸(ARA)饲料8周,麻醉处死,分离睾丸组织及附睾组织,采用气相色谱(GC)检测睾丸组织中脂肪酸的组成及含量,采用精子质量分析仪(CASA)检测附睾组织中精子形态及运动活力[精子总活力、直线运动精子活力、精子平均路径速度(VAP)、精子曲线速度(VCL)及精子直线速度(VSL)],采用苏木精-伊红(HE)染色观察睾丸组织的形态学特征,采用实时荧光定量PCR(RT-qPCR)检测2组小鼠睾丸组织中游离脂肪酸受体1(FFAR1)、FFAR2、FFAR3、FFAR4及过氧化物酶体增殖物激活受体α(PPARα)、PPARβ、PPARγ信使RNA(mRNA)的表达,采用蛋白质印迹法检测2组小鼠睾丸组织中FFAR4和PPARγ蛋白的水平。结果与WT组相比,TG组小鼠睾丸组织中ARA与二十二碳四烯酸(DTA)含量降低(P<0.05或P<0.01),亚油酸(LA)、二十碳五烯酸(EPA)、二十二碳六烯酸(DHA)含量升高(P<0.05或P<0.01);2组小鼠的精子形态及数量无明显差异,TG组小鼠精子的直线运动精子活力、VSL较WT组升高(P<0.05),精子总活力、VAP、VCL无差异(P>0.05);与WT组相比,TG组小鼠睾丸组织曲细精管内空泡较少,成熟精子增多;与WT组相比,TG组小鼠睾丸组织中FFAR4、PPARαmRNA表达上调(P<0.01或P<0.05),PPARβmRNA表达下调(P<0.05),FFAR4蛋白水平增加(P<0.05)。结论n-3 PUFAs可改善Δ15 Des转基因小鼠睾丸组织的结构和功能,其机制可能与结合FFAR4、激活下游信号分子有关。

补肾扶正汤治疗不孕不育症的价值

目的 探究分析补肾扶正汤治疗不孕不育症的临床价值。方法 选取2016年1月—2022年5月在医院接受治疗的82例不孕不育症患者作为研究对象,采用数字随机选取法平均分为对照组与试验组,对照组使用西药进行常规治疗,用药包括醋酸泼尼松片、维生素C片、维生素E片等,试验组使用补肾扶正汤治疗,方剂包括黄芪、当归、菟丝子等。治疗后比较两组女性患者的雌二醇(E_(2))、黄体生成素(LH)、卵泡刺激素(FSH)水平、子宫内膜厚度及卵泡直径;两组男性患者的精子数量及精子活动率;以及两组不良反应发生率。结果 (1)试验组女性患者雌二醇(E_(2))水平高于对照组,LH、FSH水平低于对照组,子宫内膜厚度及卵泡直径均优于对照组;(2)试验组男性患者精子数量多于对照组,精子活动率高于对照组;(3)试验组不良反应发生率低于对照组。差异均有统计学意义(P<0.05)。结论 扶正补肾汤可以调节女性体内的激素分泌水平,并显著改善子宫状态,为受孕提供更良好的基础条件。还能够提高男性精液中精子的含量和精子活动率,增加女性的受孕成功率。扶正补肾汤皆采用自然药材组成,服用后极少出现不良反应,也不会对肝肾的代谢造成负担,具有很高的安全性,是治疗不孕不孕症的首选方法之一。

男性不育患者精子线粒体膜电位及顶体酶活性检测研究

目的:探讨男性不育患者精子线粒体膜电位(MMP)及顶体酶活性检测研究。方法:选取2022年7月—2023年7月赤峰学院附属医院收治的102例男性不育患者为研究组,并依据实际情况分为弱畸精子症组、弱精子症组、畸精子症组,各34例;同时选取102例健康男性作为对照组。各组均进行顶体酶活性检测以及精子线粒体膜电位检测,对比检测结果。结果:相较于对照组,研究组的顶体酶活性、MMP更低,差异有统计学意义(P<0.05)。与弱精子症组相比,畸精子症组的MMP明显降低,弱畸精子症组的MMP、顶体酶活性显著降低,差异有统计学意义(P<0.05);与畸精子症组相比,弱畸精子症组顶体酶活性明显降低,差异有统计学意义(P<0.05)。相较于对照组,研究组的正常形态精子百分率(PNM)、精子向前运动率(PFM)更低,差异有统计学意义(P<0.05)。与弱精子症组相比,畸精子症组、弱畸精子症组的PNM显著降低,差异有统计学意义(P<0.05);与畸精子症组相比,弱畸精子症组PFM、PNM明显降低,差异有统计学意义(P<0.05)。研究组的MMP、顶体酶活性与PFM、PNM之间呈正相关性(P<0.05)。结论:在男性生育能力诊断中,顶体酶活性以及MMP检测的价值较高,检测结果和精子形态、活动力密切相关,是评定男性不育患者精子质量的重要检测指标。

Effect of Melatonin on Proliferative Activity and Apoptosis in Spermatogenic Cells in Mouse under Chemotherapy

Objective To investigate the possible protective role of melatonin on the proliferative activity, and apoptosis of germ cells in chemotherapy-induced spermiotoxicity. Methods Male adult NMRI mice were divided into four groups. Then each group was divided into two subgroups of a and b. Group A （control）： mice received vehicle （1% ethanol）for 5 d. Group B （busulfan）： mice received a single dose of 20 mg/kg busulfan. Group C （melatonin）： mice received melatonin （10 mg/kg） for 5 d. Group D （busulfan＋melatonin）： mice received a 5-day course of melatonin （10 mg/kg） following an initial dose of busulfan （20 mg/kg）. Evaluations were made using bromodeoxyuridine （BrdU） labeling and in situ TUNEL assay after 5 d （subgroup a） or 35 d （subgroup b）. Results Busulfan-treated mice both in subgroups a and b, showed a significant increase in the number of apoptotic cells （P〈0.01） and decrease in BrdU labeling index （P〈0.01） compared with controls. Melatonin in group C significantly decreased BrdU labeling index compared with the control （P〈0.01）. Melatonin in group D significantly reduced apoptosis rate and increased BrdU labeling index compared with group B. Conclusion Melatonin may have a protective effect against busulfan-induced testicular damage, partly by decreasing of apoptosis and alteration in proliferative activity of germ cells.

Cost and safety of assisted reproductive technologies for human immunodeficiency virus-1 discordant couples

Due to significant advances in the treatment of human immunodeficiency virus type-1(HIV-1), HIV-1 infection gradually has become a treatable chronic disease. Successfully treated HIV-positive individuals can have a normal life expectancy. Hence, more and more HIV-1 discordant couples in Taiwan and the rest of the world are seeking fertility assistance. Pre-treatment of highly active antiretroviral therapy(HAART) combined with sperm washing and RT-polymerase chain reaction examination for HIV-1 viral load has become the standard procedure to assist them to conceive. However,in order to reduce the transmission risk to the lowest level for the couple and to diminish the cost of health care for the insurance institutes or government, in vitro fertilization(IVF)-intracytoplasmic sperm injection(ICSI) therapy provides the ideal solution for HIV-1 discordant couples with infected men. Intrauterine insemination(IUI) theoretically introduces more than 107 times of sperm counts or semen volume to uninfected women vs IVF-ICSI. However, since some regimens of HAART may significantly decrease the sperm motility, compared to IVF-ICSI, IUI only produces 1/5 to 1/2 pregnancy rates per cycle. Given the risk of seroconversion of HIV infection which actually happens after successful treatment, IVF-ICSI for these HIV-1 seropositive men is more cost-effective and should be the first line treatment for these cases.

Physiological Roles of Platelet-activating Factor in Mammalian and Human Reproduction

This review described origination, biosynthesis and functions of platelet-activating factor （PAF） in the reproductive system of mammals and human beings. The article mainly focused on biological roles of the phospholipid mediator in sperm fertilization and embryonic implantation. As an autocrine product of sperm and embryos, PAF markedly stimulates sperm motility and fertilization and serves as a capacitation factor in a ligand-receptor manner, After fertilization, embryo-derived PAF improves its own development, especially from fertilized ova to blastocyst stage and is thought to act as an embryo growth factor in the same manner as on sperm. Its mechanism of action was also clarified. At the end, it was presented some advances in its clinical application, followed by discussion of some issues possibly concerning in its current application.

长期超低温冷冻保存对鞍带石斑鱼精子超微结构及酶活性的影响

为探究长期超低温冷冻保存中鞍带石斑鱼精子质膜、活力、超微结构及酶活性的变化,阐明影响鞍带石斑鱼精子冷冻保存质量的相关机制。实验采集2022年鞍带石斑鱼鲜精及储存时间分别为23、49和61个月的冷冻保存精液,用伊红-苯胺黑染色方法检测精子质膜完整性;用计算机辅助精子分析仪(CASA)检测精子运动参数;测量精浆和精子中琥珀酸脱氢酶(SDH)、过氧化氢酶(CAT)、谷胱甘肽还原酶(GR)、总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)和肌酸激酶(CK)共6种酶活性的变化及三磷酸腺苷(ATP)浓度变化;用扫描电镜和透射电镜观察鲜精和冻精超微结构。结果显示,伊红-苯胺黑染色后,鲜精质膜完整性最高,为83.43%±2.73%,经过超低温冷冻后,精子质膜完整性显著降低,且随着冷冻保存时间的延长而逐渐降低。CASA结果显示,鲜精活力最高,为90.47%±3.34%,经过超低温冷冻后精子活力显著降低,但长期保存23~61个月的精子活力无显著差异,精子活力保持在(63.95%±3.66%)~(68.58%±2.73%),具有稳定的活力,且鲜精与冻精之间精子平均直线运动速度(VSL)、平均曲线运动速度(VCL)和平均路径速度(VAP)均没有显著差异。精子超微结构显示,鲜精形态结构正常、线粒体排列结构规则、形态大小正常。经过超低温冷冻保存后,精子形态结构损伤明显,表现为精子头部质膜破损、细胞质外漏、细胞核膜破损、尾部鞭毛断裂或脱落等。鞍带石斑鱼精浆和精子超低温冷冻前后6种酶活性的变化及ATP含量结果显示,经过超低温冷冻后,精子内SOD、GSH-Px和CAT及ATP含量均显著降低。精浆中酶活性升高,除GR和CAT外,其余酶活性均差异显著。研究表明,长期超低温冷冻对鞍带石斑鱼精浆和精子酶活性、精子活力及精子超微结构均具有较显著的影响。本研究结果为鱼类精子冷冻损伤机理研究积累了丰富的数据,为鱼类精子长期冷冻保存提供了技术参考和评价指标。