小鼠单侧睾丸损伤后环孢素A对Fas系统的影响

目的 :研究昆明小鼠 (KM小鼠 )单侧注射冰乙酸致睾丸损伤后环孢素A(CsA)对对侧睾丸生精功能和Fas系统表达的影响。　方法 :6 0只KM小鼠随机分为 4组 :A组为对照组 ,B组为单侧睾丸损伤组 ,C组为单侧睾丸损伤后 6h切除损伤睾丸组 ,D组为单侧睾丸损伤后 6h内开始腹腔注射CsA组。 4周后取对侧附睾尾 ,计数精子及其活率 ,对侧睾丸作石蜡切片苏木精 伊红染色和免疫组化链霉素抗生物素蛋白过氧化物酶连结 (SP)法检测Fas和FasL的表达。　结果 :D组附睾尾精子和活率计数显著高于B组 (P <0 .0 5 ) ,D组的FasL和Fas较B组显著降低 (2 4 .3± 7.0vs37.8± 5 .8和 17.8± 4 .3vs32 .4± 3.6 ,P <0 .0 5 )。　结论 :KM小鼠单侧睾丸损伤后CsA可以通过抑制Fas和FasL的表达 ,降低生精细胞凋亡 ,维持生精功能的稳定     

单侧隐睾大鼠生殖股神经在对侧睾丸损害中的作用机制研究

目的 :探讨单侧隐睾大鼠模型生殖股神经在对侧睾丸损害中的作用机制。　方法 :建立单侧隐睾大鼠模型 (2 1d龄 ) ,切断该侧生殖股神经 ,12 0d后观察对侧睾丸的生精细胞凋亡变化及组织乳酸含量变化。　结果 :切断生殖股神经后 ,对侧睾丸生精细胞凋亡为 (5 .76± 0 .76 ) % ,与对照组 (17.2 8± 1.36 ) %相比明显减少 (P <0 .0 5 ) ;乳酸含量也由 (2 .19± 0 .2 4 )mmol/L下降为 (1.70± 0 .31)mmol/L(P <0 .0 5 ) ,且乳酸含量与细胞凋亡呈正相关(P <0 .0 5 )。　结论 :单侧隐睾症对侧睾丸损伤可能与其神经传导反射性血流减少引起生精细胞凋亡有关     

小睾丸组织超微结构变化与性激素相关性研究

目的 :探讨小睾丸组织超微结构变化和性激素改变及其相关性。　方法 :对 8例小睾丸病人和 12例健康成人血清进行性激素测定 ,光镜及电镜观察小睾丸组织的超微结构变化。　结果 :小睾丸病人和正常对照组血清性激素FSH、LH、T分别为 (2 1.0 5± 9.15 )IU/Lvs (6 74± 3 5 2 )IU/L、(2 2 .88± 6 .2 5 )IU/Lvs (6 6 0± 1 4 8)IU/L、(0 .30± 0 .0 4 )nmol/Lvs (17 5 5± 9 2 5 )nmol/L ,两者相比差异有显著性 (P <0 .0 1) ;精曲小管直径和管壁厚度分别为 (37.33± 6 .80 )、(10 .30± 1.82 ) μm ,与正常组的 (198 4 6± 2 9 84 )、(2 95± 0 2 0 ) μm相比 ,差异有极显著性 (P <0 .0 1) ;组织超微结构变化显著。　结论 :小睾丸组织精曲小管、生精上皮、支持细胞、界膜、间质细胞及血管均发生严重的病理改变 ,其形成可能与遗传和免疫反应有关     

大鼠附睾精子膜甘油-3-磷酸胆碱含量变化的研究

本文报道运用Percoll梯度离心技术分离纯化大鼠附睾头、体、尾各段的精子(20例),再用酶法对精子膜甘油-3-磷酸胆碱(GPC)含量依次进行测定,结果显示:大鼠附睾头、体、尾各段精子膜GPC含量分别为112.31±28.14、108.33±37.16、74.50±25.13noml/10~8精子((?)±SD).统计学分析表明:大鼠附睾头-尾、体-尾之间的精子膜GPC含量存在显著性差异(P<0.01),而头-体间差异不显著(P>0.05).提示附睾精子膜GPC含量的变化与精子成熟即精子膜的形成及受精能力的获得均有十分密切的关系.     

黄芪注射液对镉诱导大鼠精子畸形的拮抗作用

目的 :观察黄芪注射液拮抗氯化镉诱导的大鼠精子畸形作用。　方法 :将 30只SD雄性大鼠随机分成 5组 :低浓度黄芪组 (A1)、高浓度黄芪组 (A2 )、环磷酰胺组 (CP)、氯化镉组 (Cd)和对照组 (C)。预先连续 7d分别给A1组和A2 组大鼠腹腔黄芪注射液 5g/ (kg·d)和 10g/ (kg·d) ,Cd组和CP组大鼠分别腹腔注射等量蒸馏水作对照 ,之后连续 2 1dA1组、A2 组和Cd组大鼠分别同时腹腔注射氯化镉溶液 [0 .2mg/ (kg·d) ]。CP组大鼠给予 5 0mg/ (kg·d)腹腔注射染镉后第 2 2d处死大鼠 ,观察睾丸脏器系数、精子头计数、每日精子生成量、附睾尾精子计数和畸形率以及睾丸和附睾病理学。　结果 :A2 组睾丸脏器系数、睾丸精子头计数、每日精子生成量和附睾精子计数 [(5 .6 8±1.19)、(4 9.0 1± 8.78)× 10 6/g、(10 .2 5± 2 .30 )× 10 6/ (g·d)、(4 7.5 1± 2 2 .5 1)× 10 6/ml]明显高于Cd组 [(3.11± 0 .16 )、(37.5 9± 10 .6 3)× 10 6/g、(5 .31± 0 .32 )× 10 6/ (g·d)、(10 .89± 2 .4 5 )× 10 6/ml](P <0 .0 5或P <0 .0 1) ;A2 组大鼠精子畸形率 [(7.0 4± 0 .12 ) % ]明显下降 ,与Cd组 [(17.81± 1.5 5 ) % ]相比 ,差异有极显著性 (P <0 .0 1)。　结论 :黄芪注射液可拮抗氯化镉对生殖系统的损害作用 ,对防护     

无精子症患者睾丸组织病理分型与血清抑制素B关系的研究

目的:探讨无精子症患者睾丸组织病理分型与血清抑制素B(INH B)水平间的关系,了解血清INH B在评估无精子症患者睾丸生精功能状态的敏感性和特异性。方法:对83例无精子症患者进行睾丸活组织病理检查诊断,根据病理形态的不同分为:唯支持细胞综合征组(n=21)、生精功能低下组(n=20)、生精阻滞组(n=24)和生精功能基本正常组(n=18)。患者睾丸活检前分别测定其血清INH B、卵泡刺激素(FSH)、黄体生成素(LH)及睾酮(T)水平。结果:上述4组血清INH B水平分别为(20.85±18.78)、(67.25±40.98)、(73.63±25.54)和(149.48±27.92)ng/m l。INH B水平在生精阻滞组与生精功能低下组之间差异无显著性(P>0.05),其他各组间以及与上述两组血清INH B水平间差异均有极显著性(P<0.001);FSH水平在生精阻滞组与基本正常组间差异无显著性(P>0.05),其他各组间以及与上述两组血清FSH水平间差异均有显著性(P<0.05);4组血清LH及T水平之间无相关性。结论:血清INH B水平在生精小管生精功能受损时明显降低,唯支持细胞综合征者下降最为显著。血清INH B水平可直接反映睾丸生精功能的总体状态,是判断无精子症患者睾丸生精功能更有效的诊断指标。     

脊髓损伤对大鼠睾丸生精细胞iNOS、Bcl-2基因表达的影响

目的 :探讨大鼠脊髓损伤后生精细胞减少的影响因素。　方法 :6 0只SD大鼠随机分为脊髓损伤手术组(n =4 0 )和假手术组 (n =2 0 ) ,应用免疫组化S P法检测脊髓损伤大鼠术后第 2周、第 4周睾丸生精细胞iNOS、Bcl 2表达情况。　结果 :术后第 2周 ,手术组大鼠睾丸生精细胞中iNOS表达与假手术组比较显著增强 (P <0 .0 5 ) ,Bcl 2表达与假手术组比较差异无显著性。术后第 4周 ,手术组大鼠与假手术组比较 ,睾丸生精细胞中iN OS表达显著增强 (P <0 .0 5 ) ,Bcl 2表达显著性减少 (P <0 .0 5 )。　结论 :脊髓损伤后iNOS表达的增强与Bcl 2表达的减少是造成生精细胞显著减少的原因之一     

L-肉碱对糖尿病大鼠生精细胞凋亡及附睾精子数量和活动率的影响

目的:探讨L-肉碱(LC)对糖尿病(DM)大鼠生精细胞凋亡及附睾精子数量和活动率的影响。方法:24只雄性SD大鼠随机均分为3组,一组作为对照组,剩余两组分别注射链脲佐菌素(STZ,65 mg/kg)建立DM模型。建模成功后,各组大鼠分别给予如下灌胃剂量:对照组:生理盐水;DM模型组:生理盐水;LC组:300 mg/kgLC溶液,连续灌胃6周。末次给药24 h后,麻醉处死所有大鼠,分别进行附睾精子计数并检测精子活动率,流式细胞术检测各组大鼠睾丸生精细胞凋亡情况。结果:用LC治疗后的大鼠附睾头、尾精子活动率(%)分别为53.7±1.8和60.3±1.6,显著高于DM模型大鼠(分别为32.2±2.0和40.5±1.4,P<0.05),但低于对照组大鼠精子活动率63.1±2.4和68.9±1.3。与对照组附睾尾精子相对计数[(37.8±1.1)×106/100 mg]相比,DM组显著减少[(25.5±1.1)×106/100 mg],且具有统计学差异(P<0.05);LC治疗后大鼠附睾尾精子相对计数[(32.0±1.5)×106/100 mg]比DM组显著增加(P<0.05),但仍低于对照组。与对照组生精细胞凋亡率[(3.7±1.3)%]相比,DM组生精细胞凋亡率[(52.5±4.4)%]显著上升(P<0.05);经LC治疗后,LC组大鼠生精细胞凋亡率为(35.3±3.5)%,比DM组显著降低(P<0.05),但仍显著高于对照组。结论:LC(300 mg/kg)灌胃DM大鼠6周,可以减少DM大鼠生精细胞凋亡,增加附睾精子数量,提高精子活动率。     

大鼠睾丸扭转复位后附睾唾液酸含量变化及意义

目的 :探讨大鼠睾丸扭转 2h和 4h复位后 2 4h附睾唾液酸含量的变化和意义。　方法 :用 2 4只雄性SD大鼠建立左侧睾丸扭转复位模型 ,分为对照组、扭转 2h组和 4h组 ,每组 8只。 5 甲基苯二酚法检测扭转侧附睾唾液酸的含量。　结果 :睾丸扭转 2h复位后 2 4h扭转侧附睾唾液酸含量 [(2 3.385± 9.2 2 0 )mg/mgprot]改变不明显 ;睾丸扭转 4h复位后 2 4h附睾唾液酸含量 [(13.72 5± 7.80 1)mg/mgprot]下降明显 (P <0 .0 5 )。　结论 :睾丸扭转 2h复位后 2 4h附睾分泌唾液酸功能不受影响 ,扭转 4h复位后 2 4h附睾分泌唾液酸功能下降 ;附睾耐受缺血再灌注损伤的时间可能较长。     

流式细胞仪富集大鼠A1～A4型精原干细胞

目的:探讨A型精原干细胞的A1~A4亚型细胞的分离纯化方法。方法:先使用不连续密度梯度法分离A型精原细胞,再经流式细胞分选技术分选有干细胞因子受体(c-kit)表达的亚型细胞。并采用免疫组化检测c-kit在睾丸中的表达,电镜下观察分选后细胞超微结构。结果:经密度梯度分离后的精原细胞内含c-kit阳性细胞比例为(18.65±1.69)%,未经密度梯度分离的睾丸细胞内含c-kit阳性细胞比例为(3.16±0.84)%,两者比较差异有显著性(P<0.01);c-kit阳性细胞经流式细胞术分选,阳性细胞回收率(65.90±1.24)%,活性(85.60±1.14)%。结论:以c-kit为标记物,经不连续密度梯度分离初步纯化后,使用流式细胞分选技术能有效分离A型精原干细胞亚型。     

Effect of gossypol on the concentration of sodium and potassium in the rat epididymis

The effects of administration of gossypol acetic acid (7.5 mg/kg daily for 4 weeks) on the concentration of Na+ and K+ in the rat epididymis was assessed. Epididymal fluid samples, collected by micropuncture, from the caput, corpus, proximal cauda and distal cauda epididymis from gossypol-treated and control animals were analysed for Na+ and K+ concentrations. Gossypol-treated males failed to impregnate healthy females, presumably because their sperm were immotile. In gossypol-treated rats, Na+ levels decreased significantly (P less than 0.01) in the caput, corpus, proximal and distal cauda epididymis. In contrast, the K+ concentration was increased significantly (P less than 0.05) only in the caput and corpus epididymis. This altered electrolyte milieu may be responsible, to some extent, for immotility and hence infertility.     

Immunolocalization of NBC3 and NHE3 in the rat epididymis: colocalization of NBC3 and the vacuolar H+-ATPase

In the male reproductive tract, the epididymis plays an important role in mediating transepithelial bicarbonate transport and luminal acidification. In the proximal vas deferens, a significant component of luminal acidification is Na+-independent, and mediated by specific cells that possess apical vacuolar proton pumps. In contrast, luminal acidification in the cauda epididymidis is an Na+-dependent process. The specific apical Na+-dependent H+/base transport process(es) responsible for luminal acidification have not been identified. A potential clue as to the identity of these apical Na+-dependent H+/base transporter(s) is provided by similarities between the transport properties of the epididymis and the mammalian nephron. Specifically, the H+/base transport properties of caput epididymidis resemble the mammalian renal proximal tubule, whereas the distal epididymis and vas deferens have characteristics in common with renal collecting duct intercalated cells. Given the known expression of the Na+/H+ antiporter, NHE3, in the proximal tubule, and of the electroneutral sodium bicarbonate cotransporter, NBC3, in renal intercalated cells, we determined the localization of NHE3 and NBC3 in various regions of rat epididymis. NBC3 was highly expressed on the apical membrane of apical (narrow) cells in caput epididymidis, and light (clear) cells in corpus and cauda epididymidis. The number of cells expressing apical NBC3 was highest in cauda epididymidis. The localization of NBC3 in the epididymis was identical to the vacuolar H+-ATPase. The results indicate that colocalization of NBC3 and the vacuolar H+-ATPase is not restricted to kidney intercalated cells. Moreover, the close association of the two transporters appears to be a more generalized phenomenon in cells that express high levels of vacuolar H+-ATPase. Unlike NBC3, NHE3 was most highly expressed on the apical membrane of all epithelial cells in caput epididymidis, with less expression in the corpus, and no expression in the cauda. These results suggest that apical NBC3 and NHE3 potentially play an important role in mediating luminal H+/base transport in epididymis.     

Investigation of epididymal sperm maturation in the golden hamster

During passage of hamster spermatozoa through the epididymis their maturation is shown to involve changes in the sperm head, midpiece (mitochondria) and tail. The sum of these changes results in a dramatic increase in the fertilizing potential of the spermatozoa. When comparable numbers of spermatozoa from the caput or corpus epididymis were injected into one uterine horn of mature females, following ovulation induction, and spermatozoa from the cauda epididymis were injected into the contralateral horn, no fertilization was observed with caput epididymal spermatozoa, 1.7% of oocytes were fertilized by corpus epiddymal spermatozoa, whereas 79.5% fertilization was obtained with cauda epididymal spermatozoa. Total sperm numbers increased from caput to corpus to cauda [28.3 ± 12.2, 40.6 ±20.8, 1434 ±62 mihon, respectively]. The percentage of progressively motile spermatozoa increased from 27.9 ±6.4 to 33.8 ± 4.8 to 70 ± 10.7 during this passage. Viability, measured by exclusion of the dye, propidium iodide, was significantly less in spermatozoa from the cauda than from the proximal or mid-caput epididymis. The percentage of the live cells that were stained intensely by rhodamine-123 (a measure of mitochondrial membrane potential) increased during epididymal passage from 22.8 ±7.8% in the proximal caput epididymis to 57.2 ± 16.5% in the cauda epididymis. Staining with acridine orange (a measure of DNA packaging in the sperm head) indicated an increase in chromatin condensation in cauda epididymal spermatozoa, when compared to those obtained from the caput or corpus.     

Erratum

During passage of hamster spermatozoa through the epididymis their maturation is shown to involve changes in the sperm head, midpiece (mitochondria) and tail. The sum of these changes results in a dramatic increase in the fertilizing potential of the spermatozoa. When comparable numbers of spermatozoa from the caput or corpus epididymis were injected into one uterine horn of mature females, following ovulation induction, and spermatozoa from the cauda epididymis were injected into the contralateral horn, no fertilization was observed with caput epididymal spermatozoa, 1.7% of oocytes were fertilized by corpus epididymal spermatozoa, whereas 79.5% fertilization was obtained with cauda epididymal spermatozoa. Total sperm numbers increased from caput to corpus to cauda [28.3 ± 12.2, 40.6 ± 20.8, 144 ± 62 million, respectively]. The percentage of progressively motile spermatozoa increased from 27.9 ± 6.4 to 33.8 ± 4.8 to 70 ± 10.7 during this passage. Viability, measured by exclusion of the dye, propidium iodide, was significantly less in spermatozoa from the cauda than from the proximal or mid-caput epididymis. The percentage of the live cells that were stained intensely by rhodamine-123 (a measure of mitochondria1 membrane potential) increased during epididymal passage from 22.8 ± 7.8% in the proximal caput epididymis to 57.2 ± 16.5% in the cauda epididymis. Staining with acridine orange (a measure of DNA packaging in the sperm head) indicated an increase in chromatin condensation in cauda epididymal spermatozoa, when compared to those obtained from the caput or corpus.     

Sperm membrane modulation by Sapindus mukorossi during sperm maturation

Aim: To observe the alterations in the biochemical and biophysical changes in the sperm membrane during sperm maturation in male rats treated with the water extract of the fruit pericarp of S. mukorossi. Methods: Adult male Sprague-Dawley rats were gavaged the aqueous extract of the fruit pericarp of S. mukorossi at a dose of 50 mg/kg/d for 45 days. On day 46, the sperm parameters were observed in different sections of the epididymis and the sperm superoxide dismutase and the lipid peroxidation was determined and compared with the controls. The testis and epididymis were routinely prepared for histological examination under the light microscope. Results: No significant differences in the sperm number and morphology were observed between the control and treated groups. However, a significant inhibition (P<0.05-0.01) of sperm motility in the caput, corpus and cauda regions of the epididymis was seen in the treated group. No significant histopathological changes were found in the testis and epididymis. The     

Age-dependent expression of the cystatin-related epididymal spermatogenic （Cres） gene in mouse testis and epididymis

Aim： To investigate the spatial and temporal expression of the cystatin-related epididymal spermatogenic （Cres） gene in mouse testis and epididymis during postnatal development. Methods： The QuantiGene assay and indirect immunofluorescence technique were used to examine the Cres mRNA and Cres protein level in mouse testis and epididymis on postnatal days 14, 20, 22, 28, 35, 49, 70 and 420. Results： （1） In both the testis and epididymis, Cres mRNA was fast detected on day 20, then it increased gradually from day 20 to day 70, and the high expression level maintained till day 420. （2） In the testis, the Cres protein was exclusively localized to the elongating spermatids and was first detected on day 22. The number of Cres-positive spermatids increased progressively till day 49. From day 49 to day 420, the number of Cres-positive cells was almost stable. （3） The Cres protein was first detected on day 20 in the proximal caput epididymal epithelium. By day 35, the expression level of the Cres protein increased dramatically and the high level was maintained till day 420. Moreover, the luminal fluid of the midcaput epididymis was also stained Cres-positive from day 35 on. No Cres-positive staining was observed in distal caput, corpus and cauda epididymis throughout. Conclusion： The Cres gene displays a specific age-dependent expression pattern in mouse testis and epididymis on both the mRNA and protein level.     

Free and Bound Sialic Acid in Rat and Hamster Epididymal Fluid/Freie und gebundene Sialinsäure in der Nebenhodenflüssigkeit der Ratte und des Hamsters

Total, free and bound sialic acid concentrations were determined in sperm-free luminal fluid removed by micropuncture from different regions of the rat and hamster epididymides. In the rat, total sialic acid concentrations (mean +/- s.e.m.) in the proximal caput, the mid corpus and the proximal cauda were, respectively, 25.7 +/- 1.3, 23.9 +/- 1.7 and 28.8 +/- 1.9 mM compared to 4.4 +/- 0.1 mM in blood plasma. In the hamster, total sialic acid concentrations in the distal corpus, the proximal cauda and the distal cauda were, respectively, 32.9 +/- 3.8, 26.4 +/- 1.4 and 26.6 +/- 3.2 mM compared to 4.7 +/- 0.5 mM in blood plasma. Free sialic acid accounted for approximately 70-80% of total sialic acid present in the epididymal plasma of the rat. Similarly, 82% of sialic acid in the rat blood plasma was in free form. The levels of free and bound sialic acid were not changed in different regions of the rat epididymis. Unilateral ligation of the rat efferent ducts had no effect on total, free or bound sialic acid concentrations in all regions except in the proximal cauda in which a transient increase (P less than 0.01) in free sialic acid was observed on Day-3 after EDL.     

腹股沟区皮下埋藏睾丸对兔精子发生的影响

目的探讨采用腹股沟区皮下埋藏睾丸的方式修复阴囊皮肤缺损对睾丸精子发生的影响。方法普通级健康6～8月龄新西兰大白兔60只，雄性36只，雌性24只，体重2．5～2．7kg。将雄兔随机分为实验组及对照组，每组各18只。实验组动物将双侧睾丸分别移位埋藏至双侧腹股沟区皮下，以制备睾丸埋藏修复阴囊皮肤缺损模型；对照组未作处理。两组于造模后8周末时采集精液进行常规分析，每组随机取6只兔测量睾丸表面温度，另随机每组取6只兔的睾丸进行活检，应用TUNEL法检测生精细胞凋亡情况。同时将两组未采集活检的雄兔分别与雌兔一一配对喂养，观察生育情况。结果对照组精液密度为（237．3&#177;39．7）&#215;10^9／L，精子活动率为76．9％&#177;3．8％；实验组精液密度为（4．7&#177;2．7）&#215;10^9／L，精子活动率为0；两组比较差异有统计学意义（P〈0．05）。对照组的睾丸表面温度为（36．15&#177;0．64）℃，实验组为（38．02&#177;0．36）℃，两组比较差异有统计学意义（P〈0．05）。TUNEL检测示：实验组生精上皮变薄，仅存的精原细胞和初级精母见明显凋亡，凋亡指数（apoptotic index，AI）为89．69％&#177;3．76％；对照组可见散在精母细胞和精子细胞凋亡，AI为7．73％&#177;4．95％；两组比较差异有统计学意义（P〈0．05）。实验组配对后对应的母兔均未受孕生崽；对照组配对后对应母兔均受孕生崽，平均生崽数为6只；两组比较差异有统计学意义（P〈0．05）。结论与皮瓣重建阴囊一样，腹股沟区埋藏睾丸修复阴囊皮肤缺损将导致兔生殖功能障碍，主要因睾丸局部环境温度升高诱导生精细胞过度凋亡所致。     

大鼠精子在附睾成熟过程中肉毒碱量变化的研究

肉毒碱被认为是与精子在附睾中成熟有关的成熟因子。本文报告测定大鼠附睾头部,体部及尾部精子中的肉毒碱。结果表明精子在循附睾头、体、尾运行过程中,精子中肉毒碱含量逐步增加,在附睾体一头部,附睾尾一头部的精子内的肉毒碱量呈现十分显著性差异。结果提示附睾体部或体一头部交界部位可能是精子在附睾中成熟的关键部位。