吸烟对大鼠生精细胞发育影响的研究

目的:探讨吸烟对大鼠生精细胞发育的影响。方法:自制吸烟机将大鼠制成被动吸烟模型,大鼠随机分成被动吸烟组(A、B组各10只)及相应对照组(C、D组各10只),A、B组被动吸烟8周,随后处死A组及相应对照组C组大鼠;B组停止被动吸烟后与其相应对照组D组继续观察48 d后处死。流式细胞术(FCM)检测各组大鼠生精细胞周期,放免法测定血睾酮(T)、黄体生成素(LH)水平,HE染色观察睾丸组织结构变化,透射电镜观察睾丸超微结构改变。结果:与C组比较,A组大鼠精子、精子细胞[(18.76±3.58)%]和初级精母细胞[(5.71±1.18)%]明显减少(P均<0.01),而精原细胞[(55.98±5.35)%]增加(P<0.01),增殖指数降低(P<0.01)。A组大鼠生精小管壁变薄,层次减少,生精小管内精原细胞减少,精母细胞固缩。间质细胞内质网扩张脱颗粒,高尔基复合体减少,支持细胞脂滴和溶酶体增多。A组大鼠T、LH水平低于C组(P均<0.01)。B组大鼠停止被动吸烟后,精子、精子细胞、初级精母细胞比例和增殖指数上升,T、LH水平升高,但仍低于D组。结论:吸烟导致大鼠睾丸生精上皮损伤及间质细胞和支持细胞受损,同时伴有T和LH水平的下降,延缓生精细胞增殖,停止吸烟后生精功能有逐渐恢复的趋势。     

环磷酰胺干扰精原干细胞功能的初步研究

目的:初步探讨环磷酰胺对精原干细胞分化功能的干扰作用。方法:根据W istar大鼠生精细胞发育不同阶段分为1、3、9周龄组,每组24只,随机均分为实验组和对照组,实验组腹腔内注射环磷酰胺100 mg/(kg.体重),对照组注射等量生理盐水,24 h后用原位缺口末端标记法检测生精细胞凋亡。另取仅有精原干细胞的1周龄W istar雄性大鼠60只,随机均分为实验组和对照组(给药方法同上),在给药后24 h、3周、9周分别用免疫组化法检测c-K it蛋白,流式细胞仪分析细胞周期。结果:环磷酰胺干预后,1周龄实验组与对照组大鼠生精细胞凋亡差异无显著性(P>0.05),其余各周龄组实验组生精细胞凋亡均显著增高(P<0.01)。1周龄实验组大鼠睾丸在环磷酰胺处理后不同时间点的细胞周期S期细胞比例、精原细胞c-K it蛋白表达水平均明显低于对照组(P<0.01)。结论:环磷酰胺诱导精原干细胞凋亡不明显,可能更重要的是干扰精原干细胞的增殖分化功能。     

全身性非特异性免疫反应对大鼠睾丸功能的影响——睾丸形态学的变化

本实验以ＳＤ雄性大鼠为实验对象，采用血管内注射碳粒的方法，给动物造成一种全身性的非特异性免疫反应状态，以观察在此免疫反应情况下动物睾丸的生精功能有何变化。动物经每天１次共１０次的注射，实验组动物血白细胞计数（４．０９×１０４±１．２２×１０４／ｍｍ３）明显高于对照组（１．５８×１０４±０．４１×１０４／ｍｍ３，Ｐ＜０．００１）。此时实验组动物的睾丸呈不同程度的萎缩或充血，睾丸重量（０．６５１±０．１０２ｇ／１００ｇ体重）明显低于对照组（０．７９３±０．１６２ｇ，Ｐ＜０．０５）。镜下观察可见实验组动物睾丸内有的曲细精管生精细胞发生选择性脱落，生精上皮内生精细胞常呈现错位排列；另有的小管生精上皮细胞呈空竭状态或管内所有的细胞均发生固缩。此时睾丸间质内的细胞数量却明显增多。作者认为碳粒注射诱发的全身性非特异性免疫反应可以给睾丸的生精功能造成损害，这种影响可能是通过多种途径实现的。     

泛素碳端水解酶L1在邻苯二甲酸二丁酯诱导大鼠睾丸发育异常中的表达变化

目的:利用邻苯二甲酸二丁酯(DBP)诱导大鼠睾丸发育异常,验证泛素碳端水解酶L1(UCHL1)在异常睾丸与正常大鼠睾丸中的差异表达,以进一步探讨UCHL1在DBP导致的大鼠睾丸发育异常中的作用机制。方法:孕SD大鼠40只,妊娠14~18d,随机分为2组,实验组和对照组分别予DBP 800mg/(kg·d)、大豆油5ml/d灌胃,孕期第19d(GD19)和出生后22d(PND22),分别取胎鼠及仔鼠睾丸,定量定位分析UCHL1在胎鼠及仔鼠睾丸中的表达变化。结果:GD19,UCHL1在实验组的相对表达量为0.075±0.02(n=10),对照组为0.150±0.02(n=10),2组差异有显著性(P<0.05),UCHL1在实验组中比正常对照组下降50%;而PND22,UCHL1于实验组隐睾睾丸中的相对表达量为0.344±0.03(n=10),实验组非隐睾睾丸中为0.326±0.02(n=10),对照组为0.322±0.02(n=10),3组间无明显统计学差异(P>0.05)。UCHL1主要定位于睾丸发育时期的精原细胞,初级精母细胞和次级精母细胞的胞质与胞核中。结论:DBP在染毒期间影响了睾丸生精细胞UCHL1的表达,影响了泛素-蛋白酶体系的平衡,从而导致生精小管的变薄,各层生精细胞数目减少的睾丸发育异常现象。     

苯甲酸雌二醇对大鼠睾丸组织发育的影响

目的:研究苯甲酸雌二醇(E2B)对雄性大鼠睾丸组织发育的影响。方法:选取新生雄性SD大鼠40只,随机均分为实验组和对照组,在生后1d分别皮下注射E2B(0.2mg/5g体重)、等体积的溶剂玉米油,分别于生后14、21、28、42、56d取睾丸并称重。测量头段及尾段生精小管管径(TD)及上皮高度(SEH),并计算头段及尾段TD/SEH比值、尾段SEH/头段SEH比值,分析生精小管内生精细胞发育状况。结果:各年龄段实验组大鼠睾丸重量显著下降(P<0.01),睾丸始终停留在腹腔。睾丸网内始终有液体潴留,从出生后21d开始,实验组大鼠头段睾丸的生精小管的TD/SEH比值明显大于对照组(P<0.01),尾段SEH/头段SEH比值也显著增加(P<0.01);实验组生精细胞发育明显迟滞于对照组(P<0.01)。结论:E2B可以造成雄性大鼠睾丸内液体潴留、隐睾,从而阻滞睾丸组织的正常发育。     

营养性肥胖对青春期雄性大鼠睾丸生精细胞凋亡的影响

目的:探讨营养性肥胖对青春期雄性大鼠睾丸生精细胞凋亡的影响。方法:健康雄性W istar大鼠40只,随机分为两组,每组20只,对照组给予普通饲料喂养,高脂组用高脂、高热量饲料喂养建立营养性肥胖大鼠模型,10周末处死大鼠摘取睾丸。全自动生化分析仪(ACA)检测外周血总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)和低密度脂蛋白胆固醇(LDL-C);光镜观察睾丸组织病理学改变;TUNEL检测睾丸组织凋亡细胞;免疫组化法检测Bc l-2和Bax蛋白的分布和表达;RT-PCR法检测睾丸组织Bc l-2 mRNA和Bax mRNA的表达。结果:高脂组TC、TG、HDL-C和LDL-C(5.17±0.17、1.18±0.09、1.76±0.11、5.08±0.18)较对照组(1.38±0.12、0.39±0.05、0.97±0.07、0.75±0.06)显著升高(P<0.05);高脂组生精细胞凋亡指数(37.17±2.74)较对照组(5.16±0.81)显著升高(P<0.01),凋亡细胞以精原细胞和精母细胞为主;高脂组Bax蛋白和Bax mRNA(153.26±8.74、1.08±0.12)表达较对照组(101.81±6.14、0.37±0.04)明显升高(P<0.01);高脂组Bc l-2蛋白和Bc l-2 mRNA(139.26±7.21、0.46±0.05)表达较对照组(159.37±8.96、1.05±0.11)明显降低(P<0.01)。结论:营养性肥胖诱导大鼠睾丸生精细胞凋亡增多,其机制可能与Bc l-2表达降低和Bax表达升高相关。     

皮瓣重建兔阴囊诱导精原细胞凋亡与热休克蛋白70表达

目的:探讨皮瓣重建阴囊后睾丸表面的温度变化、生精细胞凋亡及热休克蛋白70(HSP70)的表达。方法:以健康育龄期新西兰大白兔为实验动物,雄性24只、雌性12只。将雄兔随机分成实验组12只,对照组12只。采用温度计埋藏法测量实验组阴囊内睾丸表面的温度,手术切除实验组动物阴囊,建立皮瓣重建阴囊动物模型,对照组未作处理。动物模型建立后第8周末对照组随机取6只动物进行睾丸活检,实验组随机取6只动物测量睾丸表面温度后行睾丸活检。睾丸组织行HE染色、原位末端标记(TUNEL)法检测生精细胞凋亡、免疫组化法结合图象分析仪检测睾丸生精细胞HSP70的表达。模型建立后第8周末,将两组未行睾丸活检的各6只动物分别与雌兔配对喂养观察生育情况。结果:实验组模型建立前睾丸表面温度为(36.0±0.3)℃,模型建立后第8周末睾丸表面温度为(38.1±0.6)℃(P<0.05)。模型建立后第8周末实验组睾丸生精小管中生精细胞的凋亡指数(AI)为(71.85±2.69)%,对照组为(7.73±4.95)%(P<0.05)。对照组HSP70主要在圆形精子细胞中表达,实验组主要表达在精原细胞。配对喂养结果显示,实验组没有生育,对照组生育幼兔(6.0±1.3)只(P<0.05)。结论:皮瓣重建家兔阴囊后睾丸局部温度升高诱导生精细胞凋亡增加,动物丧失生育能力。HSP70参与保护皮瓣重建阴囊诱导的精原细胞凋亡。     

抗FSH自身抗体对大鼠睾丸生精功能影响的初步研究

目的:建立抗FSH自身抗体大鼠模型,同时研究抗FSH自身抗体对雄性大鼠生精功能的影响。方法:30只SD大鼠(21d龄),随机分为实验组、对照组,各15只。合成大鼠FSHβ亚基上一段特异性氨基酸序列(18肽),将合成多肽与钥孔戚血蓝素(keyholelimpethemocyanin,KLH)偶联,免疫SD大鼠,作为实验组。对照组大鼠用KLH免疫。初次免疫之后,每隔2周加强免疫1次,共7次,于第3次加强免疫后1周(即,免疫第49d)眼眶取血测定血清抗体效价,之后每隔两周测一次抗体效价,至实验结束。并于免疫第77d、91d和105d时,分别处死实验组和对照组大鼠各5只,用光镜和电镜观察大鼠睾丸生精小管结构和附睾精子结构的变化,计数附睾尾精子数量及肿胀精子百分率,并用ELISA法检测大鼠血清T水平。结果:免疫第49d后,实验组大鼠血清抗18肽-KLH抗体效价为1∶200,免疫第63d后,抗体效价达到1∶400,之后抗体效价一直维持在1∶400。免疫第91d后,实验组大鼠附睾尾肿胀精子百分率显著低于对照组(60.4±6.23vs50.60±3.05,P<0.05);免疫第105d后,实验组大鼠生精小管内生精细胞数量及管腔内精子数量均减少,且附睾尾精子数量(46.08±6.56vs32.53±3.41)和肿胀精子百分率(60.60±5.86vs48.60±3.85)均显著低于对照组(P<0.05),而血清T水平显著高于对照组(P<0.05)。结论:抗FSH自身抗体可能会引起睾丸生精障碍。     

青春期前己烯雌酚暴露对SD大鼠睾丸发育及功能的影响

目的:研究青春期前己烯雌酚(DES)暴露对SD大鼠睾丸发育及功能的影响。方法:21日龄雄性SD大鼠90只,随机分为DES0.01、0.1、1.0、10.0μg/(kg.d)4个实验组和1个对照组(分别为Da、Db、Dc、Dd和C组,每组n=18)。于青春期前,即出生后第22d(postnatalday22,PND22)～35d(PND35),实验组每日皮下注射相应剂量的DES,共14d,对照组仅注射溶媒(玉米油)。观察各组大鼠睾丸下降时间。于青春期晚期(PND50)、性成熟后(PND64)和成年期(PND130)分3批(每批n=6)处死各组大鼠取材,测定睾丸重量,观察比较睾丸组织形态学变化,分析PND130大鼠附睾尾精子质量。结果:C、Da、Db、Dc和Dd组睾丸下降时间分别为PND26.17±1.94、26.83±1.47、28.68±1.03、33.50±1.87和41.50±2.74,其中Db、Dc和Dd组较C组明显延迟(P<0.05或P<0.01)。PND50时,C、Da、Db、Dc和Dd组单侧睾丸重量分别为(1.38±0.10)、(1.38±0.12)、(1.30±0.14)、(0.86±0.18)g和(0.73±0.27)g,其中Dc和Dd组较C组显著减轻(P<0.01);与C组比较,Db组仅有少数生精小管生精上皮中的细胞数目稍减少,Dc和Dd组生精小管发育较差、生精上皮中细胞数目减少、精子发生阻滞、间质细胞发育幼稚,其程度随DES暴露剂量增加而加重。PND64时,C、Da、Db、Dc和Dd组单侧睾丸重量分别为(1.60±0.06)、(1.62±0.11)、(1.58±0.08)、(1.47±0.10)g和(0.99±0.37)g,其中Dc和Dd组较C组显著减轻(P<0.05或P<0.01);Dc和Dd组睾丸组织形态学改变与PND50时类似,但总体较PND50有所改善。PND130时,各实验组与对照组比较,单侧睾丸重量差异无统计学意义(P>0.05),睾丸组织形态学改变难以鉴别出明显差异;C、Da、Db、Dc和Dd组大鼠附睾尾精子密度分别为(73.00±16.90)、(68.00±19.67)、(68.67±12.15)、(35.17±15.64)×106/ml和(19.13±5.17)×106/ml,其中Dc和Dd组精子密度较C组明显降低(P<0.01);与C组比较,Dd组精子活动率下降(P<0.01),Db、Dc和Dd组a级精子比例降低(P<0.05或P<0.01),Dd组b级精子比例降低(P<0.01)。结论:青春期前小剂量DES[0.01μg/(kg.d)×14d]暴露对SD大鼠睾丸发育及功能无明显影响,较大剂量DES[1.0～10.0μg/(kg.d)×14d]暴露对大鼠睾丸发育及功能有明显的近期(PND50和PND64)和较远期(PND130)毒性作用,该毒性作用随DES的暴露剂量增加而加重,随鼠龄增长而逐渐减退,其机制可能与间质细胞和支持细胞的发育及功能受损相关。     

干细胞因子在2,5己二酮所致大鼠睾丸生精障碍模型中的表达

目的　探讨膜型及可溶型干细胞因子 (SCF)的表达与精子生成的关系。方法　 5只成年雄性SD大鼠水饲 2 ,5己二酮 5周 ,建立睾丸生精障碍模型 ;逆转录 多聚酶链反应 (RT PCR)和MIAGS 10 0 0凝胶电泳成像系统分析睾丸组织中两种SCF的表达及其表达水平 ,并与对照组比较。结果　实验组大鼠睾丸重 ( 0 .5 3± 0 .0 5 ) g ,与对照组 ( 1.6 7± 0 .0 7) g比较明显萎缩 ( P <0 .0 1) ;组织切片显示实验组大鼠睾丸生精上皮萎缩 ,仅有支持细胞和精原细胞 ,无成熟的精细胞 ;实验组中膜型和可溶型SCF的相对蛋白含量分别为 2 8.7和 73 .0 ,对照组为 83 .4和 75 .1,显示膜型SCF含量在实验组明显降低 (P <0 .0 1) ,而可溶型SCF无明显变化 (P >0 .1)。结论　膜型SCF表达水平降低可能在 2 ,5己二酮所致大鼠睾丸生精障碍中起重要作用。     

低温联合地塞米松对大鼠睾丸扭转复位后eNOS表达及生精细胞凋亡的影响

目的:探讨低温联合地塞米松对睾丸扭转复位后的保护作用,以及对eNOS表达及生精细胞凋亡的影响。方法:将80只青春期SD大鼠随机分为4组,每组20只。4组大鼠分别扭转左侧睾丸720°2 h,建立单侧睾丸扭转模型,随后各组做如下处理,A组:常温+生理盐水、B组:低温+生理盐水、C组:低温+地塞米松、D组:常温+地塞米松;术后48 h采集睾丸,通过HE染色光镜观察睾丸组织病理学改变、免疫组化法检测eNOS表达、TUNEL法检测睾丸生精细胞凋亡。结果:HE染色光镜下见4组大鼠扭转侧睾丸组织均有不同程度损伤,其中A组睾丸损伤最明显,其余3组扭转侧睾丸得到不同程度保护;睾丸组织eNOS免疫组化检测结果:A组扭转侧(左侧)睾丸组织阳性细胞数及阳性细胞着色强度明显强于B、C、D 3组,差异具有显著性(P<0.05、P<0.01、P<0.01);凋亡细胞染色:细胞核呈深棕黄色或棕褐色,A组扭转侧(左侧)睾丸可见大量生精细胞凋亡,凋亡指数AI(31.12±4.68)明显高于B组(16.58±6.22)(P<0.05)及C(8.60±1.15)、D组(13.52±3.06)(P<0.01)。结论:睾丸扭转复位后的缺血再灌注损伤可导致生精细胞凋亡增加、睾丸生殖能力下降;应用低温联合地塞米松能显著增强睾丸组织的抗损伤能力,较好地保护了扭转复位后睾丸的生精功能。     

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。     

L-肉碱在奥硝唑所致大鼠睾丸和附睾损伤中的保护作用

目的:探讨L-肉碱(LC)在奥硝唑(ORN)所致大鼠附睾和睾丸损伤中的的保护作用。方法:40只雄性SD大鼠(200～230g)随机均分为5组:①A组:给予0.5%的羧甲基纤维素钠(溶剂)灌胃;②B组:每天给予400mg/kgORN灌胃;③C组:每天给予800mg/kgORN灌胃;④D组:每天给予[ORN(400mg/kg)+LC(100mg/kg)]灌胃;⑤E组:每天给予[ORN(800mg/kg)+LC(100mg/kg)]灌胃。上述各组均连续灌胃20d,末次给药24h后,所有大鼠麻醉后处死,分别取睾丸、附睾,进行称重和HE染色,计算睾丸、附睾系数并观察睾丸和附睾病理组织学改变。结果:①与A组相比,B组睾丸、附睾系数明显降低(P<0.05);而C组睾丸、附睾系数为极显著性降低(P<0.01);D组与A组相比无差异,E组与A组相比有极显著性差异(P<0.01);②HE染色显示,与A组相比,B组睾丸生精小管内各级生精细胞排列基本整齐,部分生精小管管腔内有脱落的生精细胞,附睾管腔中精子数目下降,有时可见散在的生精细胞;C组大鼠睾丸生精小管管腔内均可见坏死脱落的生精细胞,附睾管腔中精子数目明显减少,且有较多的非精子细胞成分。D组睾丸生精小管无明显改变,附睾管腔中精子数目也未见明显下降;E组睾丸生精小管管腔内精子数目减少,可见坏死脱落的生精细胞,附睾腔中精子数目明显减少,并伴有较多的非精子细胞成分。结论:奥硝唑(ORN)可导致雄性大鼠附睾和睾丸病理组织学改变,LC对ORN引起大鼠附睾和睾丸损伤具有一定的保护作用。     

154例无精子症睾丸活检组织病理和定量组织学观察

本研究对154例无精子症睾丸活检进行了组织病理观察,并在此基础上应用QTM-720图象分析仪对这些活检组织进行了定量组织学研究,测出(1)曲细精管平均直径,(2)曲细精管上皮平均厚度,(3)曲细精管总截面积与测试面积之比,(4)每一测试面积中曲细精管截面数。各病理组间上述四个参数有显著差异,与光镜观察结果相符。     

Morphological responses of the rabbit testis to ischemic/reperfusion injury due to torsion

OBJECTIVE: To investigate the morphological effects of free radical injury on spermatogenic cells in both testes of the rabbit experimental model of testicular torsion. MATERIALS AND METHODS: The left testes of 8 peripubertal NZW rabbits (3-6 months) were subjected to 0, 15, 30, 60, 90, 120, and 180 min of ischemia by applying a clamp to the spermatic cord, followed by reperfusion. Another set of 8 rabbits was subjected to 60 min of ischemia and administered antioxidants (acetylsalicylic acid, ascorbic acid, allopurinol, quercetin, superoxide dismutase) before reperfusion. Both testes of 4 animals per group were harvested at 24 h and the remaining 4 at 3 months. Johnsen scores for spermatogenic activity and other changes were assessed histologically and these were compared with testicular malondialdehyde (MDA), a measure of free radical damage, assayed on testicular homogenates using the thiobarbiturate method. RESULTS: In the 24-hour reperfusion group, apoptotic bodies and giant cells were more prominent in the seminiferous tubules of the left testes compared to the right, and were maximal after 90 min. In the 3-month reperfusion group, giant cells were absent, and apoptotic bodies were reduced in both testes. Testicular MDA showed an increase only in the left testes in the 24-hour reperfusion group, while the 3-month group showed increased MDA levels in both testes, but more on the left. The Johnsen score fell only to 8.0 in the left testes in the 24-hour reperfusion group, but dropped to 2.3 in the 3-month reperfusion group. Only in the 3-month reperfusion group, did antioxidant-treated animals show a fall in Johnsen scores in the left testes, regardless of the type of antioxidant. CONCLUSION: These findings confirm a role for reactive oxygen species (ROS) in damage to spermatogenic cells in both the ipsilateral and contralateral testes following torsion, with longer term effects in the torted testis. Currently available antioxidants do not provide any significant long-term protection against morphological damage to the testis by ROS generated in testicular torsion.     

未成熟大鼠睾丸单侧扭转后对健侧血流和组织学的影响

目的:观察未成熟大鼠睾丸单侧扭转后对健侧睾丸血流供应和组织学的影响,并比较不同处理方法的疗效。方法:建立Wistar3周龄大鼠左侧睾丸扭转模型,分别建立对照组、扭转组、扭转复位组和扭转切除组,每组10只。彩色多普勒测量各组术前、术后8h(即扭转复位或切除术后2h)、12h、24h、72h右侧睾丸动脉收缩期最大血流速度,并于对照组和扭转组术后2h,扭转复位组和扭转切除组第1次术后12h各取2只大鼠右侧睾丸进行组织病理学观察。各组喂养至9周龄时分别取右侧睾丸进行组织学观察及检测各组大鼠右侧睾丸的生精小管直径(STD)、生精上皮细胞计数(CMSE)和睾丸活检评分(TBS)。结果:①未成熟睾丸左侧扭转后,右侧睾丸的血供呈持续性增加。②扭转组、扭转复位组和扭转切除组右侧睾丸均有不同程度的间质水肿和超微结构改变。③9周龄时扭转组、扭转切除组右侧睾丸重量均较对照组显著增加(P<0.01);各组大鼠STD、CMSE、TBS均无显著性差异(P>0.05)。结论:未成熟大鼠睾丸单侧扭转后可引起对侧睾丸的血供增加和组织学改变,轻微损伤后扭转复位和睾丸切除预后效果相似。     

Decreased testicular expression of cAMP response element modulator (CREM) in rat with varicocele

This study examines the hypothesis that varicocele would impair the testicular expression of cAMP response element modulator (CREM) in experimental rats. Thirty (30) rats were selected at random, of 20 were operated as varicocele's group; of 10 were for sham-operation as controlled group, testes were removed, fixed and stained in three months. Makler Score was adopted to analyze the bore, limitan's thickness, layer number of cell, the degree of cell's maturity and average score in 200 and 100 seminiferous tubulae, respectively. CREM, HSP60 was determinated by hybridization in situ, the difference between two groups were compared. In varicocele group, seminiferous tubule's bore was decreased (101 +/- 2.2) vs 146 +/- 4.1), limitan became thicker (3.5 +/- 0.1 vs 1.9 +/- 0.2), cell's layer number was reduced (3.0 +/- 0.2 vs 5.5 +/- 0.1), cell's maturity turned to disturbance (3.6 +/- 0.3 vs 4.9 +/- 0.1), the average score was lower than controlled group (8.5 +/- 0.6 vs 16.0 +/- 1.2), they had significant differences (P < 0.001). The testicular CREM expression was significantly lower in varicocele group than in controlled one (VG2.0 +/- 0.32, SoG3.90 +/- 0.32) (p < 0.001), which was located in spermatogenous cell to spermatocyte stage. However, HSP60 expression in VG was higher than in SoG (3.85 +/- 0.3 versus 2.1 +/- 0.32) with significant differences (P < 0.001), the expression located in spermatid. Varicocele could lead to lower testicular CREM expression and breeding sperm functional lesion.     

The correlation between serum epidermal growth factor/testicular epidermal growth factor receptor and spermatogenesis in rat

<正>Objective: To investigate the correlation between epidermal growth factor (EGF)/testicular epidermal growth factor receptor (EGF-R) and spermatogenesis in rat. Methods: Forty mature male Spraque-Dauley (SD) rats were randomly assigned to four groups, ten rats in each: sham operation group (SOG), sialoadenectomy group(SG), sialoade-nectomy group with injection of EGF (0. 25 μg·kg-1·d-1, SG-EGF Ⅰ) and sialoadenectomy group with injection of EGF (0. 50 μg·kg-1·d-1 , SG-EGF Ⅱ). The rats were routinely feed, and blood and testes were obtained on the 48th day after the operation. Serum EGF concentrations were determined by radioimmunoassay (RIA) , expression of EGF-R in testes was examined by the immunohistochemical method, and the spermatogenesis was pathologically checked. Results: Serum EGF levels in SG-EGFIand SG decreased significantly when compared with those of SOG (P<0. 05 and P< 0. 01, respectively). The testicular function of spermatogenesis showed a moderate to severe impairment in SG. The expression of EGF-R in Leydig cells decreased in SG(P<0. 05). The two dosage groups of EGF replacement had different effects. There were no significant differences of EGF-R expression in testicular germ cells, Sertoli cells and Leydig cells in SOG, SG-EGFⅠand SG-EGFⅡ(P>0. 05). Conclusion: EGF may play an important role in the regulation of spermatogenesis. Serum EGF concentration and high expression of EGF-R in Leydig cells have a positive correlation with spermatogenic function of the testes.     

急性弓形虫感染对雄性小鼠睾丸生精功能影响的初步研究

目的:探讨弓形虫急性感染对雄性小鼠睾丸生精功能的影响。方法:将26只成年雄性小鼠随机均分为感染组和正常对照组;感染组腹腔注射0.3ml1×103/ml弓形虫速殖子,感染小鼠睾丸,正常组腹腔注射等量生理盐水,两组睾丸制成直接印片及病理切片,观察小鼠生精细胞的病理变化及弓形虫侵入生精细胞情况。同时比较两组睾丸乳酸脱氢酶同工酶、精子密度、精子活动率、精子畸形率。结果:感染组睾丸乳酸脱氢酶同工酶、精子密度、精子活动率、精子畸形率分别为53.19±18.04、(15.01±2.42)×106/ml、(8.26±2.57)%、(17.69±11.91)%,正常组为68.71±17.79、(23.87±6.66)×106/ml、(13.21±2.82)%、(11.30±6.60)%,两组比较差异均有显著性(P<0.05)。结论:弓形虫感染对雄性小鼠睾丸生精功能有一定的影响。     

Investigation on the histopathological effects of thyroidectomy on the seminiferous tubules of immature and adult rats

INTRODUCTION: This study aimed to investigate the histopathological effects of thyroidectomy on both immature and adult rat testes. MATERIALS AND METHODS: Male albino Wistar rats, 4 weeks old and weighing between 45 and 55 g, were used for this study. The experimental groups were as follows: 2-week control group (group I); 2-week thyroidectomy group (group II); 4-week control group (group III); 4-week thyroidectomy group (group IV); 6-week control group (group V), and 6-week thyroidectomy group (group VI). The control groups included both sham-operated and untreated rats. In groups II, IV and VI, total thyroidectomy was performed under ether anesthesia in all rats at 4 weeks of age. The rats were killed in the 2nd, 4th and 6th weeks, respectively, following the thyroidectomy. The testes of each animal were evaluated histologically. RESULTS: In group II, spermatogenesis progressed to meiosis but round spermatids were found to be decreased and pachytene spermatocytes were observed to be increased when compared to group I. Giant pachytene spermatocytes were seen. There were also many degenerated cells of intermediate origin in the seminiferous epithelium. In groups IV and VI, spermatogonia and primary spermatocytes were normal in appearance, but there was widespread degeneration of the other spermatogenic cells. In addition, some closed lumina covered by degenerated and dead cells were observed. In group II, the mean outer diameter, luminal diameter and area occupied by seminiferous epithelium decreased by 19.74, 32.18, and 28.12%, respectively. In group IV, these data decreased by 23.9, 16.52, and 48.5%, respectively, and in group VI, by 21.10, 19.76 and 40.29%, respectively, when compared with the control groups. These data were statistically significant (p < 0.001). CONCLUSIONS: Thyroid hormones could have a marked influence on the seminiferous tubules of both immature and adult rats, and their permanent lack results in a depression in seminiferous tubule growth as shown by the reduced outer and luminal diameters and area occupied by the seminiferous epithelium, which could give rise to degenerative changes in the spermatogenic cells of thyroidectomized rats. In addition, all these changes could also result from both the inability of Sertoli cells to support spermatogenic cells and the diminished levels of GH and FSH.