刺五加提取物体外对精子运动参数的影响

目的 研究3种不同的刺五加提取物体外对精子运动参数的影响。方法 由14例弱精子症患者通过手淫获得并经上游优化处理的精子，与3种不同的中药刺五加提取物一起孵化0h、0．5h、2h后，采用计算机辅助的精子分析系统(CASA)检测精子的运动参数。结果 刺五加水层及饱和正丁醇层提取物在体外能显著提高人精子的活率、前向性运动百分率、直线运动速度(VsL)和曲线运动速度(VCL)。结论 中药刺五加水层和饱和正丁醇层提取物在体外能显著改善人精子的运动功能。     

刺五加水提物体外对弱精子症患者精子运动参数的影响

目的:研究不同浓度的刺五加水提物体外对弱精子症患者精子运动的影响,探讨其可能的作用机制。方法:将35例弱精子症患者经手淫法获得并通过上游优化处理的精子,与不同浓度刺五加水提物共同孵育30、60、120、180 m in后,应用计算机辅助精子分析系统(CASA)观察不同浓度(2.5、5、10、20 g/L)刺五加水提物对人精子各运动参数的作用。结果:不同浓度刺五加水提物能明显提高弱精子症患者精子的运动能力,在浓度为5 g/L和10 g/L时,精子活率(MOT)、前向运动精子百分率、曲线运动速度(VCL)、直线运动速度(VSL)和平均路径速度(VAP)与空白对照组相比,差异均有显著性(P<0.05)。结论:刺五加水提物在体外能明显改善弱精子症患者精子运动能力,其最佳浓度为10 g/L。     

磷脂酰肌醇3激酶抑制剂体外对弱精子症患者精子运动参数的影响

目的:研究磷脂酰肌醇3激酶(PI3K)的特异性抑制剂LY294002在体外对弱精子症患者精子运动参数的影响,并探讨其可能的作用机制。方法:弱精子症患者和精液参数正常男性各10例,手淫法留取精液,经上游优化处理后的精子与不同浓度的LY294002孵育10、30、60 m in后,采用计算机辅助精液分析系统检测精子的运动参数。结果:LY294002在体外能显著提高弱精子症患者精子的活动率、前向性运动精子百分率和直线运动速度、平均路径运动速度。结论:LY294002在体外显著增强弱精子症患者精子的运动功能。     

rhGM-CSF体外对人精子运动参数的影响

目的观察体外添加不同浓度的重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)对人精子运动参数的影响,探讨其在精子运动中的作用机制。方法健康生育男性和弱精子症患者各10例手淫取精,经简易上游优化处理后的精子与不同浓度rhGM-CSF溶液孵育10min、30min、60min后,采用计算机辅助的精液分析系统检测精子各项运动参数的变化。结果对精液运动参数正常的标本,1ng/ml和10ng/ml的rhGM-CSF溶液能显著提高精子的活率、前向性运动百分率,在10ng/ml时平均直线运动速度也有显著提高。对弱精子症患者精液标本,1ng/ml和10ng/ml的rhGM-CSF溶液能显著提高精子的前向运动百分率和直线运动速度,10ng/ml的rhGM-CSF对精子活率和平均路径速度也有显著性提高。两类精子的曲线运动速度均无显著性变化。结论1ng/ml～10ng/ml浓度范围的rhGM-CSF体外能显著改善精子的运动功能。     

金黄色葡萄球菌和表皮葡萄球菌体外对人精子运动功能的影响

目的 :研究金黄色葡萄球菌 (SA)和表皮葡萄球菌 (SE)体外对人精子的运动功能有无直接影响。　方法 :将泌尿生殖系感染病人中分离培养的SA和SE制成活菌悬液 ,体外与 10例健康成年男性手淫获得并经上游优化处理的精子孵育。按细菌 /精子比 5 0∶1分别孵育 0、2、4h后 ,采用计算机辅助的精子分析系统检测人精子活率、运动参数 (前向性运动百分率、直线速度、曲线速度、平均路径速度 )和精子形态及凝集情况。　结果 :SA与精子体外孵育 2、4h后 ,精子活率和运动参数明显降低 (P <0 .0 5 )。SE体外对精子的活率和运动功能则无明显影响。　结论 :SA与精子的比例为 5 0∶1,体外孵育 2h后 ,能显著降低精子活率和抑制精子运动功能。SE对精子运动功能无直接影响。     

氰戊菊酯体外对大鼠精子运动能力的影响

目的:观察氰戊菊酯(Fen)对大鼠精子运动能力的直接作用。方法:收集7只健康成年雄性SD大鼠附睾尾精子,用Fen进行体外染毒,剂量分别为1、4、16、64μmol/L,以Fen溶剂(二甲亚砜)作为对照组。在染毒1、2、4 h后用计算机辅助精子分析(CASA)系统对精子运动速度[曲线运动速度(VCL)、直线运动速度(VSL)、平均路径速度(VAP)、鞭打频率(BCF)]和运动方式[前向性(STR)、直线性(LIN)]进行检测,观察Fen对离体大鼠精子运动能力的影响。结果:Fen染毒1、2 h后,可见64μmol/L组的VSL、BCF、LIN和STR与对照组相比差异有显著性(P<0.01或P<0.05)。染毒4 h后,除上述指标外,VCL在16、64μmol/L组与对照组相比也呈现出明显的下降(P<0.01)。时间-效应关系分析显示,Fen(16、64μmol/L)染毒4 h组与染毒1 h组相比,VCL和STR显著下降(P<0.01或P<0.05)。结论:Fen可作用于大鼠附睾尾精子,并对大鼠精子运动有直接毒性效应。     

白念珠菌体外感染对人精子运动和超微结构的影响

目的: 体外研究白念珠菌(Ca)对人精子的运动功能有无直接影响并观察精子超微结构变化,对其影响机制作初步探讨。　方法: 将从念珠菌性阴道炎患者的分泌物中分离纯化的Ca制成活菌悬液,对 10例健康青年男性手淫获得并经上游优化处理的精子进行体外感染。按细菌与精子比例不同分成A组 ( 1∶1 )、B组 ( 1∶10 )、C组(1∶100)、D组(1∶1 000)、E组(1∶10 000)、F组(空白对照)分别孵育,于 0、1、2、4h取样,计算机辅助精子分析系统(CASA)检测人精子运动参数(前向性运动百分率、直线速度、曲线速度、平均路径速度、头部侧摆幅度 )、精子存活率、精子形态及凝集情况。透射电镜观察孵育 4h后的各组精子的超微结构变化。　结果: Ca与精子体外孵育后,前向运动百分率所受影响最大,且与菌体密度及时间密切相关;其他参数与对照组相比也相继出现显著性差异。观察到精子黏附于菌体和凝集现象。精子超微结构产生变化:精子核空泡增多,顶体破裂,质膜破损,线粒体排列紊乱。　结论: Ca体外可显著降低精子存活率和抑制精子运动功能,其机制可能与Ca对人精子的黏附作用和超微结构的损伤有关。     

淋病奈瑟菌体外对人精子运动参数的影响

目的 :阐明淋病奈瑟菌体外对人精子的运动参数有无直接影响。　方法 :将淋病奈瑟菌制成活菌悬液 ,在体外与生育男性经上游优化处理的精子孵育。细菌 /精子比为 5 0∶1,分别孵育 0、2、4h ,采用计算机辅助的精子分析系统检测人精子运动参数 (前向性运动百分率、直线速度、曲线速度、平均路径速度、直线性、前向性 )。　结果 :细菌与精子的比例为 5 0∶1,孵育 0、2和 4h后 ,人精子运动参数无明显变化。　结论 :淋病奈瑟菌与精子的比例为5 0∶1,孵育 4h内 ,淋病奈瑟菌体外对人精子运动参数无直接影响     

两种精子分析系统的比较研究

目的:评估自动化精子质量分析仪SQA-V与计算机辅助精液分析(CASA)系统之间各主要参数的差异性及其在不育和生育男性精液质量分析中的应用。方法:用SQA-V和CASA系统分别检测12例正常生育者和73例不育患者新鲜精液标本,分别分析精子密度、精子活动率、活动精子浓度、精子活动力指数、精子头侧向位移、鞭打频率、精子曲线运动速度、精子前向运动速度、平均路径速度、直线性及前向性运动速度等参数,针对两者进行相关性分析。结果:不育组和生育组男性各项指标间存在显著差异,两种分析系统检测的精子密度之间(r=0.58,P<0.01)、活动精子浓度之间(r=0.75,P<0.01)、平均精子速度(SQA-V)和精子路径速度(CASA)之间(r=0.59,P<0.01)具有较好的一致性,SQA-V的活动精子指数(SMI)和CASA的前向性、曲线运动速度、精子运动的直线性、前向运动速度、平均路径速度、鞭打频率等参数具有显著相关性(P<0.05)。结论:SQA-V和CASA系统比较,在各主要参数上具有较好的一致性,能够反映临床不同组群患者精液的差异性。     

人参皂甙Rb1体外对人精子运动参数的影响

目的研究人参皂甙Rb1体外对正常男性精子运动的影响,探讨其可能的作用机制。方法将18例正常男性经手淫获得的精子,通过上游优化法处理后,与10μmol/L人参皂甙Rb1共同孵育0．5h、1h、2h、3h、4h、5h,应用计算机辅助的精子分析系统(CASA),观察随时间延长人参皂甙Rb1对人精子各运动参数的作用。结果随着时间延长,精子活动能力均有不同程度下降；10μmol/L人参皂甙Rb1能明显提高正常男性精子的运动能力(A B)%、精子存活率(Mot)、前向运动精子百分率、曲线运动速度(VCL)、直线运动速度(VSL)和平均路径速度(VAP),与空白对照组相比,差异均有显著性(P＜0．05)。结论随着时间延长,低浓度人参皂甙Rb1在体外能明显提高止常男性精子的存活率和运动能力,本研究为人参皂甙Rb1的临床应用提供了斯的实验基础。     

Effects of several Chinese herbal aqueous extracts on human sperm motility in vitro

The effects of six kinds of aqueous extracts of Chinese herbal medicine (Astragalus membranaceus, Acanthopanacis senticosi, Panax genseng and Ophiopogon japonicus, P. genseng and Aconitum carmichaeli, Salviae miltiorrhiae, Polyporus umbellatus polysaccharide) on sperm motility characteristics of 30 infertile male volunteers were studied in vitro with a computer-assisted sperm analysis at 15, 60 and 180 min after incubated with the drugs. The results showed that per cent viability, number of progressive motile spermatozoa, curvilinear velocity, average path velocity and amplitude of lateral head displacement were significantly enhanced by A. membranaceus (P < 0.05 or < 0.01), per cent viability, average path velocity and amplitude of lateral head displacement were significantly enhanced by A. senticosi (P < 0.05), but all the above were not affected by P. genseng and O. japonicus, P. genseng and A. carmichaeli, S. miltiorrhiae and P. umbellatus polysaccharide. It is suggested that A. membranaceus and A. senticosi can enhance the motility of human spermatozoa in vitro.     

The effects of caffeine and theophylline on the triple adenosine triphosphatase enzyme activities of spermatozoa from males with oligoasthenospermia

The triple ATPase activities of washed spermatozoa of oligoasthenospermic men (but not of normals) were enhanced by the addition of caffeine and theophylline, which are known to have a stimulating effect on sperm motility. The biochemical mechanism of action of caffeine and theophylline on sperm homeostasis is discussed.     

梗阻性无精子症患者睾丸精子左旋肉碱培养后关键基因表达量的变化研究

目的:探讨肉碱对人精子活力的影响及其对男性不育的治疗作用。方法:以梗阻性无精子症患者睾丸穿刺取得的精子为研究对象,通过普通培养液、培养液中添加100mmol/L及250mmol/L肉碱进行培养,比较培养前后精子活动率的变化,以及用RT-PCR检测男性生殖细胞特异性基因Vasa、Dazl、Acr、Prm1及ATPase6.0表达量的变化,探讨左旋肉碱与有关精子发生和成熟过程的重要功能基因表达的关系。结果:培养24～72h,添加100mmol/L的左旋肉碱培养液运动精子数明显较未添加和添加250mmol/L组高(P<0.01);100mmol/L左旋肉碱培养液组中典型的快速前向运动精子数明显较多,巴氏染色法观察其具有正常的精子结构,RT-PCR检测表明100mmol/L的肉碱可提高精子中Acr、Prm1、Dazl及ATPase6.0基因的表达量,而250mmol/L的肉碱组Dazl、Acr、Prm1基因的表达减少。结论:合适浓度的左旋肉碱可通过上调一些生殖相关基因的表达使得睾丸精子培养后活力增强,有利于对睾丸穿刺患者行单精子注射时精子的选择。如培养液中肉碱浓度过高,可能由于肉碱的毒性作用,反而使得这些基因的表达量减少。     

The application of pentoxifylline in the stimulation of sperm motion in men undergoing electroejaculation

The ability of pentoxifylline to stimulate the motion characteristics of antegrade and retrograde sperm collected at the time of electroejaculation with a rectal probe was assessed in six neurologically impaired men. Before electroejaculation, the bladder was rinsed and instilled with 20 to 30 ml Ham's F-10 medium. Washed sperm were incubated with various doses (0, 0.1, 1, and 3 mmol/L) of pentoxifylline. Video sequences were recorded at intervals from 0 to 4.5 hours and analyzed for sperm motion parameters using manual and computer-assisted semen analysis. The results were compared with equimolar concentrations of caffeine. Both pentoxifylline and caffeine demonstrated a dose-dependent stimulation of percent motility and other motion parameters. A maximal stimulation of two-fold to three-fold for both percent motility and curvilinear velocity, and 30% to 60% for straight line velocity was observed after incubation under these conditions. A significant increase in mean linearity was observed in samples incubated with 0.1 mmol/L pentoxifylline at 1.5 hours. Significant lateral head displacement was not observed at any time point. Two couples underwent gamete intrafallopian transfer (GIFT) in conjunction with this electroejaculation sperm stimulation procedure, and one has since delivered a normal child. These studies show that pentoxifylline stimulation can improve the movement characteristics of asthenospermic sperm from neurologically impaired men. Such sperm stimulation techniques do not affect the fertilization process and may improve the chances for conception in some cases of male-factor infertility.     

Effect of pentoxifylline on motility and membrane integrity of cryopreserved human spermatozoa

The purpose of this study was to examine the effects of pentoxifylline used before and after semen cryopreservation-thawing on sperm motility and membrane integrity. Twenty-four semen samples were split into four equal aliquots. Aliquots were incubated at 37 degrees C for 30 min, followed by cryopreservation with TEST-yolk freezing medium using slow programmable freezing protocol. After 2 weeks the sperm samples were thawed, washed twice in Quinn's Sperm Washing Medium (modified HTF with 5.0 mg/mL Human Albumin) and incubated at 37 degrees C for 30 min. Aliquots were treated by adding 3 mmol/L pentoxifylline to: (1) fresh sperm samples during incubation period prior to cryopreservation, (2) sperm samples as a supplement to the cryoprotectant prior to cryopreservation, and (3) thawed sperm samples during incubation period. One aliquot received no treatment (control group). The addition of 3 mmol/L pentoxifylline to fresh semen during incubation period prior to cryopreservation significantly decreased progressive and total motility compared with controls. However, the addition of 3 mmol/L pentoxifylline to cryopreserved semen after thawing significantly increased progressive and total motility compared with controls. After post-thaw, no differences in motion characteristics between sperm samples treated by adding 3 mmol/L pentoxifylline as a supplement to the cryoprotectant and control groups were observed. Post-thaw hypoosmotic swelling (HOS) test scores did not improve with the addition of pentoxifylline compared with the control group. It is concluded that pentoxifylline enhanced post-thaw motility of cryopreserved human spermatozoa when added after thawing. No improvement was found by freezing sperm with pentoxifylline.     

Inhibition of alkaline phosphatase activity of boar semen by pentoxifylline,caffeine, and theophylline

Methyl xanthines have been used frequently as additives to sperm suspensions in order to improve sperm characteristics. The mechanism of action on spermatozoa is generally assumed to be inhibition of sperm phosphodiesterase activity, resulting in elevation of complementary adenosine monophosphate levels in spermatozoa. The present study was designed to examine the effect of methyl xanthines (pentoxifylline, caffeine, and theophylline) on another important enzyme system, alkaline phosphatase, in boar seminal plasma and spermatozoa. Inhibition of sperm alkaline phosphatase could be distinguished from that of seminal plasma by a paradoxical stimulation by pentoxifylline at lower pH values in spermatozoa. Among the three methyl xanthines, theophylline exhibited the most dramatic inhibition of alkaline phosphatase activity and substrate inhibition was observed with increasing concentrations. Each methyl xanthine had a different action on alkaline phosphatase activity at lower pH; theophylline showed the highest inhibition, caffeine inhibition was not related to pH, and pentoxifylline did not inhibit alkaline phosphatase of seminal plasma and, in fact, it stimulated its activity (or that of a phosphatase with lower pH optimum) in spermatozoa. These results indicate another possible mechanism of action of methyl xanthines on sperm and are in agreement with data indicating that methyl xanthines are not specific inhibitors of sperm phosphodiesterase, because clearly, they inhibit alkaline phosphatase activity as well.     

Evolution of total antioxidant status in a model of acute renal insufficiency in rats

BACKGROUND: Accurate estimation of the Total Antioxidant Status (TAS) in the myoglobinuric acute renal failure (ARF) is necessary because its pathogenesis is believed to be mediated, at least in part, by the development of oxidative stress resulting from the generation of oxygen free radicals and reduced antioxidant defense system. The purpose of this study is to examine the TAS 24 and 72 h after glycerol injection in a model of myoglobinuric-ARF. EXPERIMENTAL DESIGN: The study was conduced in 28 Sprague-Dawley rats. In group 1 (n = 7) rats were placed into individual metabolic cages and deprived of water during 24 h. afterwards an intramuscular injection of glycerol was administrated (50% vol/vol in sterile saline) 10 mg/100 g of body weight and 24 h later blood samples were collected for biochemical measurements (urea, creatinine, creatine-kinase, and TAS levels). In group 2 (n = 7), rats followed the same conditions than group 1 ones but blood samples were collected 72 h after glycerol injection. In groups 3 (n = 7) and 4 (n = 7) rats didn't receive glycerol injection, and blood samples were collected within 24 and 72 h respectively after they were placed into metabolic cages. RESULTS: In groups 1 and 2 we observed a renal function decrease, with higher serum levels of urea and creatinine in group 2 (urea levels: 269 +/- 16 mg/dL vs. 586 +/- 147 mg/dL; p < 0.001. Creatinine levels: 2.8 +/- 0.2 mg/dL vs. 5.8 +/- 0.7 mg/dL; p < 0.001). TAS levels in groups 2, 3, and 4 were similar, but in group 1 was significantly lower (group 1: 0.81 +/- 0.2 mmol/L; group 2: 1.3 +/- 0.1 mmol/L; group 3: 1.2 +/- 0.3 mmol/L, and group 4: 1.2 +/- 0.2 mmol/L; p < 0.005). CONCLUSION: In the model of glycerol induced myoglobinuric-ARF we observed a decrease of serum TAS level within 24 h with spontaneous recuperation 72 h after.     

The Effect of Caffeine on Guinea Pig Epididymal Spermatozoa: Motility and Fertilizing Capacity

Guinea pig epididymal spermatozoa, when taken from different segments of the epididymis, are known to differ markedly in their in vitro motility, metabolism and morphology. In this report, the fertilizing capacity of epididymal spermatozoa isolated from proximal and distal segments and treated with caffeine was tested. Twenty six guinea pig sows were inseminated 16–26 h after parturition with isolated epididymal spermatozoa. In 15 cases the spermatozoa were treated with caffeine (10 mM), and 11 cases served as controls. Sperm characteristics (mean ± SE) were: proximal sperm; concentration 48 ± 3.9 millions/ml, vitality 94 ± 0.9%, motility 42 ± 5.3%. Distal sperm: concentration 161 ± 21 million/ml, vitality 94 ± 0.9% and motility 86 ± 3.0%. Caffeine increased significantly the motility of proximal spermatozoa (by 59.5%). These suspensions of spermatozoa were used for intraperitoneal artificial insemination.
The impregnation rate of untreated proximal sperm was zero (0 out of 4), compared with 83% (5 out of 6) for the caffeine treated sperm. The impregnation rate of untreated distal segment sperm was 42.8% (3 out of 7) compared with 100% (9 out of 9) for caffeine treated sperm.     

氯化锶激活对小鼠体细胞核移植胚胎卵裂率和囊胚率的影响

目的:建立氯化锶(SrCl2)诱导小鼠体细胞核移植胚胎激活的最佳方法。方法:首先构建及鉴定小鼠核移植胚胎,在注核完成后,使用以下实验激活核移植胚胎:实验1,采用5 mmol/L和10 mmol/L SrCl2处理核移植胚胎1～8 h,观察时间对卵裂率和囊胚率的影响;实验2,在4、6 h采用1～20 mmol/L多种浓度SrCl2处理核移植胚胎,观察不同浓度对卵裂率和囊胚率的影响;实验3,研究10 mmol/L SrCl2配合不同培养液激活核移植胚胎的效果;实验4,观察10 mmol/L SrCl2联合蛋白激酶抑制剂6-DMAP、细胞松驰素(CB)对体外培育核移植胚胎的影响。结果:研究发现当浓度一致时(5 mmol/L),6 h组卵裂率(38.9%)与1 h组(6.7%)、2 h组(22.8%)、3 h组(22.8%)、4 h组(25.6%)比较差异均有显著性(P<0.05),但与5 h组(28.9%)、7 h组(34.4%)、8 h组(28.9%)均无显著差异(P>0.05);当时间固定时(6 h),SrCl2浓度为10 mmol/L获得的卵裂率和囊胚率(68.9%和7.2%)较高,较1 mmol/L组(28.3%和0%)、2.5 mmol/L组(35.6%和0%)、5 mmol/L组(37.8%和1.1%)、7.5 mmol/L组(60.6%和2.2%)、15 mmol/L组(51.7%和1.1%)和20 mmol/L组(41.7%和1.1%)均高(P<0.05);不同成分培养液实验显示,含Ca2+和Mg2+KSOM组的胚胎卵裂率较低,仅为27.8%,与不含Ca2+组(69.4%)、不含Ca2+/Mg2+组(66.1%)和添加EDTA组(68.3%)比较差异均显著(P<0.05);虽然SrCl2联合各培养液对核移植胚胎体外发育影响不大,但SrCl2联合CB组囊胚细胞计数(45.40±2.23)较未联合组(30.15±1.12)、联合6-DMAP组(34.95±1.38)、联合6-DMAP+CB组(37.45±1.43)均显著增多(P<0.05)。结论:10 mmol/L SrCl2单独处理6 h或联合CB均能较好地激活小鼠体细胞核移植胚胎。