线粒体功能与精子活力

精子活力是衡量精液质量和男性生育力的一个重要指标,在男性不育患者中约19％为精子活力低下所致。精子运动所需能量来自线粒体呼吸链的氧化磷酸化,各种原因导致的线粒体结构和功能改变,如膜电位的降低,酶活性或表达量异常以及线粒体DNA（mtDNA）的突变或缺失等均可导致线粒体能量合成障碍,精子活力降低。对近年有关线粒体功能与精子活力相关性研究进展综述,旨在探讨精子活力低下的发病机制以及临床治疗弱精子症的可能途径。     

线粒体功能与精子活力

精子活力是衡量精液质量和男性生育力的一个重要指标,在男性不育患者中约19%为精子活力低下所致。精子运动所需能量来自线粒体呼吸链的氧化磷酸化,各种原因导致的线粒体结构和功能改变,如膜电位的降低,酶活性或表达量异常以及线粒体DNA（mtDNA）的突变或缺失等均可导致线粒体能量合成障碍,精子活力降低。对近年有关线粒体功能与精子活力相关性研究进展综述,旨在探讨精子活力低下的发病机制以及临床治疗弱精子症的可能途径。     

人卵泡液蛋白质组学的研究进展

卵泡液是由卵母细胞、颗粒细胞和膜细胞等共同分泌的一种含多种蛋白质的生物液,其作为卵母细胞生长的微环境或培养基,直接影响到卵泡的发育和卵母细胞的成熟。过去十几年,对卵泡液蛋白质组学的研究主要集中于新蛋白筛选及疾病相关蛋白等的识别上。本文从卵泡液新蛋白的筛选、年龄相关生物学标志鉴定、确定卵泡成熟标志、疾病相关标志的筛选和体外受精（IVF）成功的标志等5方面介绍近年卵泡液蛋白质组学的相关研究。筛选和鉴定某些生殖和发育相关的关键蛋白,并可能成为卵泡液中的生物标志,可用于预测卵母细胞的质量、受精及胚胎发育潜能。     

芹菜汁对小鼠精液参数及毒性作用的影响

目的:探讨芹菜汁对小鼠精液参数以及毒性作用的影响,为芹菜的进一步开发利用提供一定依据。方法:将小鼠按体重随机分为7d、14d和28d3个时间组,再将时间组按体重随机分为4组(对照组、低剂量组、中剂量组和高剂量组),每天定时给对照组0.3ml生理盐水灌胃,剂量组给0.3ml不同浓度的芹菜汁灌胃,分别于各时间段结束后处死动物。应用WLJY-9000型伟力彩色精子质量检测系统测定精子运动参数。结果:7d组、14d组中高剂量组和中剂量组精子密度与相应对照组比较差异均有统计学意义(P<0.05)。中剂量组:VCL14d组和28d组分别和7d组比较有统计学意义(P<0.01),VSL7d组和14d组比较有统计学意义(P<0.05);高剂量组:VCL、MAD、WOB7d组和14d组比较均有统计学意义(P<0.05)。结论:芹菜汁能够影响小鼠精子运动参数,降低精子密度。     

卵泡液干细胞因子水平对卵母细胞发育、受精和卵裂影响的实验研究

目的探讨卵泡液中干细胞因子(SCF)水平对卵母细胞发育、受精和卵裂的影响。方法采用酶联免疫吸附法(ELISA)检测68例体外受精-胚胎移植患者卵泡液中SCF水平,分析其与卵母细胞发育、受精和卵裂的关系。结果卵泡液SCF水平为526±58ng/L,显著高于血清SCF水平(389±31ng/L)(P<0.01)。含成熟卵母细胞的卵泡液SCF水平为540±62ng/L,高于含未成熟卵母细胞者(515±55ng/L)(P<0.05);受精卵的SCF水平(538±61ng/L)高于未受精卵(519±56ng/L)(P<0.05);Ⅰ级和Ⅱ级胚胎的SCF水平(536±59ng/L、531±60ng/L)高于Ⅲ级和Ⅳ级胚胎(522±56ng/L、520±55ng/L)(P<0.05)。结论卵泡液SCF可促进卵母细胞发育,提高受精卵发育潜能。     

Optimum Measurement Time for Human Sperm Velocity Determination

The optimal time-duration required by spermatozoa to travel distances from which their average velocity can be accurately determined was investigated. For this purpose, 180 motile spermatozoa, from 12 seminal specimens, were photographed while being illuminated by six light pulses, given at 1-sec intervals. From these photographs, distances traveled by each spermatozoon were measured over identical time periods. Most individual spermatozoa showed slight deviation of their velocity when measured sequentially for 5 sec. Nevertheless, when mean sperm velocity of grouped spermatozoa were determined from 1- or 5-sec measurement time, the results were almost identical. This was attributed to a pure statistical law, that is, the mean deviation decreases as the number of spermatozoa in the sample increases. It was concluded that the sperm velocity of a certain specimen can be determined properly using a 1-sec measurement time. An extended period of time does not seem to contribute to the accuracy of sperm velocity determination.     

卵泡液中细胞因子与卵泡发育及妊娠结局的相关性

随着体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVF-ET)技术水平的日益提高,卵巢微环境中调节卵母细胞发育的细胞因子的作用受到广泛关注。细胞因子作为一类具有广泛生物学活性的激素样多肽调控生殖过程,它们通过调节卵巢内分泌或旁分泌,进而影响卵泡成熟、胚胎发育及胚泡着床等环节来参与生殖调节。研究表明不孕可能与卵泡液中某些细胞因子(如血管内皮生长因子、粒细胞集落刺激因子、活性氧簇、胰岛素样生长因子、白细胞介素、白血病抑制因子等)的异常表达引发的卵泡成熟障碍有关。因而,加强卵泡液中细胞因子的研究将有助于提高IVF-ET的临床妊娠率。     

多囊卵巢综合征患者卵泡液中6种miRNAs表达的检测

目的：分析多囊卵巢综合征（PCOS）患者卵泡液外泌体中miRNA-9、miRNA-18b、miRNA-19、miRNA-24、miRNA-199和miRNA-320水平,初步探讨其在PCOS发生、发展中的潜在意义。方法：48例接受体外受精-胚胎移植（IVF-ET）治疗的不孕症患者,分为PCOS组（23例）和对照组（25例）,留取其卵泡液。采用超速离心法提取卵泡液外泌体,分别用外泌体粒径分析（NTA）、蛋白质印迹（Western blotting）和透射电镜验证外泌体。通过实时荧光聚合酶链式反应（PCR）法检测卵泡液和外泌体miRNA-9,miRNA-18b,miRNA-19,miRNA-24,miRNA-199,miRNA-320的表达情况。结果：卵泡液中PCOS组miRNA-199的表达水平高于对照组（5.42±0.80 vs. 1.02±0.94,P＜0.01）,miRNA-320的表达水平低于对照组（0.47±0.06 vs. 1.23±0.09,P＜0.01）;2组间miRNA-9,miRNA-18b,miRNA-19和miRNA-24的表达水平差异无统计学意义（P＞0.05）。在外泌体中,PCOS组miRNA-9和miRNA-18b的表达水平高于对照组（8.94±0.54 vs. 1.01±0.97,2.66±0.19 vs. 1.12±0.06,均P＜0.01）,miRNA-199和miRNA-320的表达水平则低于对照组（0.43±0.11 vs. 1.22±0.07,0.53±0.09 vs. 1.18±0.15,均P＜0.01）。结论：PCOS患者卵泡液和外泌体存在miRNA-9,miRNA-18b和miRNA-199的表达,可能与PCOS发生、发展相关。     

Characterization of Hyperactivated Motility by Human Spermatozoa During Capacitation: Comparison of Fertile and Oligozoospermic Sperm Populations

Suspensions of capacitating human spermatozoa were analyzed for potential hyperactivated movements using videomicrographic methods. Analysis was carried out on aliquots of 22 sperm suspensions, which were proved fertile several hours later during human in vitro fertilization. After approximately 3 h of capacitation, 22.1% of the fertile spermatozoa displayed motility patterns designated as hyperactivated. Over 80% of these hyperactivated spermatozoa moved with a wide-amplitude, two-dimensional whiplash pattern, displaying marked lateral displacement of the head. Only 8.4% of capacitating spermatozoa from oligozoospermic patients showed these hyperactivated movements. The incidence of hyperactivated movements by fertile and oligozoospermic spermatozoa could be significantly increased after exposure to various motility stimulants. The clinical significance of hyperactivation as a functional assay of fertilizing capacity is discussed.     

精子活动力与线粒体代谢的研究进展

弱精子症或精子无力症是一种由于精子活动力缺陷所引起的男性不育,约占男性不育症的19％,已成为男性不育的主要诱因之一.精子运动所需能量基本来自其线粒体呼吸链的氧化磷酸化,而任何与线粒体代谢相关的因素,如氧化应激、细胞凋亡及Ca2+循环等均可对精子活动力产生影响.综述近年来对精子活动力与其线粒体代谢的相关性及机制研究的进展.     

Significance of Adenosinetriphosphate in Human Sperm as Clinical Parameter

Washed human spermatozoa from 21 individuals with an average motility of 60% (quality index 277 ± 16) had an endogenous ATP generation of 7.5 ± 3.4 nmole/10⁸spermatozoa. The ATP concentrations in spermatozoa from 16 patients with severely impaired motility of 26% (quality index 98 ± 13) was 16.9 ± 9.9 nmole/10⁸spermatozoa. There was no correlation between ATP content and motility in either group. Sperm penetration into blood serum type AB, Rh-positive, was evaluated using a capillary tube penetration test. The penetration was graded with a maximum score of 14. Spermatozoa with an initial motility of 60% reached a score of 10 ± 0.7. After addition of 20 mmole of ATP the score was significantly improved to 13 ± 0.3. Compared with these results spermatozoa with an average motility of 26% reached a score of 4 ± 1. Exogenous ATP (20 mmole) increased the score to 8 ± 1.0. In both groups reduced glutathione had a negative effect.

Human spermatozoa with high and low motility are capable to synthesize ATP. A dysfunction of the phosphorylating particles in the mitochondria appears not to be associated with low sperm motility.     

Effect of Ace-Inhibitors,Calmodulin Antagonists,Acetylcholine Receptor Blocking,and Alpha Receptor Blocking Agents on Motility of Human Sperm

The purpose of this study was to investigate the influence of several pharmacological compounds on the motility and velocity of washed human spermatozoa. Results were evaluated by multiple exposure photography and computer-aided picture analysis. The motility-inhibiting effect of the antifertility drug gossypol was confirmed. Gossypol proved to be a potent inhibitor of the angiotensin converting enzyme (ACE) detectable in high concentrations in seminal plasma. However, human sperm motility was not inhibited during incubation with two other specific ACE-inhibitors (captopril, enalapril). On the contrary, high concentrations of captopril even showed a slight motility-stimulating effect. These results indicate no direct involvement of ACE in the regulation of sperm motility but suggest a direct interaction of gossypol with the plasma membrane of spermatozoa. To clarify whether or not gossypol blocks membranous ion transport, the effect of well-defined ion transport blocking agents on sperm motility was investigated. It was determined that the acetylcholine receptor blocker alpha-bungarotoxin and trifluoperazin, a specific calmodulin antagonist, inhibit sperm motility completely. Since stimulation of sperm motility by captopril may be due to an alpha-mimetic action of this compound, the influence of two alpha receptor blockers (bromocriptine, lisuride) on sperm motility was studied. Although lisuride inhibited sperm motility completely, bromocriptine revealed no influence. A temporary and reversible intervention with membrane transport processes could be a suitable way to regulate human sperm motility and male fertility.     

体外操作对人精子运动参数及细胞内活性氧簇的影响

目的：研究体外物理操作（离心以及微量加样器抽吸）对人精子运动参数以及细胞内活性氧簇（ROS）的影响,旨在优化人精子体外处理方法。方法：7例精液常规参数正常的标本,采用不同离心力（200 g,600 g）及离心时间（5 min,15 min）以及不同微量加样器抽吸次数（2,6,10次）进行处理,测试精子运动参数和细胞内ROS的变化。结果：①活动精子百分比以及活精子细胞内ROS水平主要受到离心时间的影响（P＜0.05）;前向运动精子百分比（PR）以及平均运动速率（VAP）均不受离心力及离心时间的影响（P＞0.05）。单因素方差分析多重比较结果提示,当离心时间为5 min以上时精子活动力显著下降,而ROS水平显著增加（P＜0.05）。②加样器抽吸2次以上可致人精子活动力显著下降（P＜0.05）,抽吸6次以上可导致人精子PR及VAP显著降低（P＜0.05）;但抽吸操作并未显著影响精子细胞内ROS的水平。结论：在人精子体外操作时应尽量缩短离心时间并控制加样器抽吸次数从而保证正常的精子运动参数,也应尽量缩短离心时间从而减少体外操作所致的细胞内ROS升高。     

用计算机辅助精子分析系统测定TCDD对大鼠精子运动能力的影响

利用计算机辅助的精子分析系统（ＣＡＳＡ）研究了２,３,７,８—四氯二苯－Ｐ－二口恶口英（ＴＣＤＤ）对大鼠精子运动能力的影响。给２１天龄幼鼠腹腔注射ＴＣＤＤ０．１、１．０和５．０μｇ／ｋｇ,对照组注射等量体积的溶媒。在性成熟后（９０天）,用扩散法收集附睾尾精子,测定其运行速度（ＶＣＬ、ＶＳＬ、ＶＡＰ）、运动方式（ＳＴＲ、ＬＩＮ、ＢＣＦ、ＡＬＨ、ＭＡＤ等）及活动精子的比率（Ｍｏｔ％）。结果表明,ＴＣＤＤ１．０μｇ／ｋｇ组,精子的运动速度明显低于对照组（ＶＣＬＰ＜０．０５;ＶＡＰ、ＶＳＬ、ＢＣＦＰ＜０．０１）,０．５μｇ／ｋｇ时精子的运动速度、及前向性和直线性运动性能均明显降低（Ｐ＜０．０１）,活动精子的比率也明显低于对照组（Ｐ＜０．０１）。本研究结果为ＴＣＤＤ的男性生殖毒理学研究,以及ＣＡＳＡ在男性生殖毒理学研究中的应用提供了进一步参考依据     

血清和卵泡液中睾酮水平与体外受精-胚胎移植结局的关系

目的：探讨体外受精-胚胎移植（IVF-ET）治疗时血清和卵泡液中睾酮浓度与IVF-ET结局的关系。方法：选择2011年5—8月在成都中医药大学第二附属医院生殖中心接受IVF-ET或胞浆内单精子注射（ICSI）-ET治疗的患者43例,检测患者基础血清及控制性超促排卵（COH）时血清和卵泡液中睾酮水平。①按获卵数分3组,低反应（L）组（1～5个）、正常反应（M）组（6～15个）、高反应（H）组（＞15个）,分析各组血清和卵泡液中睾酮浓度及各组受精率和妊娠率。②按临床妊娠情况分为妊娠组与未妊娠组,比较2组间睾酮的差异。结果：①不同获卵数3组间卵泡液中睾酮水平和基础睾酮水平差异无统计学意义（P＞0.05）;人绒毛膜促性腺激素（hCG）日3组血清睾酮水平比较差异有统计学意义（P＜0.05）,其中H组高于M组及L组（P分别为0.010和0.001）;COH中其余时间点睾酮水平差异无统计学意义（P＞0.05）。②3组受精率、优质胚胎率差异无统计学意义（P＞0.05）;临床妊娠率比较差异有统计学意义（P＜0.05）。③妊娠组各阶段血清睾酮水平仅在取卵日时高于未妊娠组,差异有统计学意义（P＜0.05）;2组卵泡液中睾酮差异无统计学意义（P＞0.05）。结论：IVF-ET治疗时基础血清睾酮水平不能预测卵巢反应和妊娠结局,但hCG日睾酮水平可能预测卵巢反应;取卵日血清睾酮与妊娠有关;卵泡液中睾酮对妊娠结局无显著影响。     

镉对大鼠精子运动能力影响的体外实验研究

赖皮地精子运动能力的直接作用。方法应用计算机辅助精子分析（ＣＡＳＡ）系统了氯化镉对离体大鼠精子运动能力的影响。利用扩散法收集健康成年雄性ＳＤ大鼠附睾尾精子,用氯化镉进行体外染毒,染毒浓度分别为０．５、２．０、８．０ｍｇ／Ｌ（以镉计）。分别在染毒１ｈ和２ｈ后应用ＣＡＤＡ系统对精子运动速度（运动速度、直线运动速度、平均路径速度、鞭打频率）和运动方式（前向性,直线性等）进行检测。「结果」在染毒１ｈ后染毒     

Effect of Caffeine on Human Sperm Penetration into Zona-Free Hamster ova

The effect of caffeine on spermatozoal ability to penetrate zona-free hamster ova was examined on fresh and frozen-thawed semen samples. The mean motility of 10 fresh semen samples incubated with caffeine significantly increased from 29% to 35%. Sperm penetration into zona-free hamster ova did not differ between the control group and the specimens to which caffeine was added. The same effect of caffeine on sperm motility and hamster ova penetration was noted in the frozen-thawed sperm samples. Motility was enhanced by 21%, but hamster ova penetration did not significantly change. The increase in sperm motility caused by caffeine does not change the fertilizing ability of fresh and frozen-thawed human sperm.     

丝裂原活化蛋白激酶信号转导通路与精子活动力

丝裂原活化蛋白激酶(MAPK)属于丝/苏氨酸蛋白激酶,激活后可参与调节细胞生长、发育、分化、凋亡等一系列生命活动。MAPK转导通路的一些组成部分,如细胞因子:白细胞介素6(IL-6)、成纤维细胞生长因子(FGF);蛋白激酶C(PKC);细胞外信号调节激酶(ERK)和p38及底物精子微管蛋白等直接或间接调节精子的活动力。MAPK信号通路的研究对探讨弱精子症发生的分子机制,寻求弱精子症新治疗途径有意义。     

顶体蛋白酶抑制剂KF-950对人精子功能的影响

本文研究了顶体蛋白酶抑制剂KF-950对人精子功能的影响。结果表明:KF-950对人精子的作用与其浓度和作用时间有关。经≥0.21ml KF-950处理,随着时间的延长,精子活率和活力明显下降;精子膜功能有不同程度的损害,且对精子头部膜的损害较其对尾部膜的损害严重,顶体脱落率及已发生顶体反应的精子死亡率明显上升。KF-950可通过破坏精子膜而影响精子的功能。     

Effect of L-Carnitine on Motility and Acrosome Reaction of Human Spermatozoa

L-carnitine added to the suspension medium decreases the glucose-sustained progressive motility of human spermatozoa. Addition of 20 mM L-carnitine to the capacitation medium causes an inhibition of the occurrence of the acrosome reaction parallel to a viability enhancement and negligible changes of the cellular content of ATP. The cellular efflux of glutamate-oxaloacetate transaminase was also inhibited by L-carnitine. A possible role of L-carnitine on membrane stability and metabolism of spermatozoa is briefly discussed.