青春期前己烯雌酚暴露对大鼠性成熟后生精细胞凋亡的影响及其机制初探

目的:研究青春期前己烯雌酚(d iethylstilbestrol,DES)暴露对SD大鼠性成熟后睾丸生精细胞凋亡的影响并初步探讨其机制。方法:30只21日龄雄性SD大鼠,随机分为DES 0.01、0.1、1.0、10.0μg/(kg.d)4个实验组和1个对照组(编码为ADa、ADb、AD c、ADd和AC组,每组6只)。于青春期前[出生后第22 d(postnatal day 22,PND22)至35 d(PND35)],实验组每日皮下注射相应剂量的DES,对照组仅注射溶媒。于大鼠性成熟后(PND 64)处死各组大鼠切取双侧睾丸,采用TUNEL法检测大鼠睾丸生精细胞凋亡,用免疫组化方法检测凋亡相关蛋白Bc l-2和Bax在生精细胞中的表达。结果:与对照组相比,ADa组大鼠性成熟后生精细胞凋亡无明显变化,ADb、AD c和ADd 3组生精细胞凋亡增加,且随DES暴露剂量增加而有增加趋势。AC、ADa组生精细胞Bax相对弱表达而Bc l-2强表达,伴随DES暴露剂量增加,Bax表达逐渐增强而Bc l-2表达逐渐减弱,ADd组Bax强表达而Bc l-2弱表达。结论:青春期前较大剂量DES暴露可使大鼠性成熟后睾丸生精细胞凋亡增加,且随DES暴露剂量增加而有加强趋势。凋亡相关蛋白Bax和Bc l-2参与青春期前DES暴露所致的生精细胞凋亡过程。     

大鼠脊髓损伤急性期睾丸生精细胞凋亡及Fas和Bcl-2蛋白表达的改变

目的 研究脊髓损伤急性期对成年大鼠睾丸生精功能障碍的影响及其机制.方法 80只SD雄性大鼠随机分为脊髓损伤组(40只)和脊髓非损伤组(40只),采用原位细胞凋亡TUNEL法检测脊髓损伤后第1、2、4、8、1 6天睾丸生精细胞的形态及数量变化.采用免疫组化SABC法检测大鼠脊髓损伤后第1、2、4、8、16天睾丸生精细胞Fas、Bcl-2表达情况.结果 大鼠脊髓损伤后睾丸TUNEL标记阳性凋亡细胞第4天开始出现,多表达于初级、次级精母细胞及精原细胞,至第16d,随时间点推移阳性凋亡细胞数逐渐增多.脊髓损伤后第4～16d损伤组睾丸生精细胞Fas表达与非损伤组比较显著增强(P＜0.05),Bcl-2表达损伤组与非损伤组差别无显著性(P＞0.05).结论 大鼠脊髓损伤急性期存在睾丸生精细胞凋亡现象,睾丸生精功能障碍与Fas高表达密切相关,和Bcl-2表达无明显相关性.     

生精冲剂对实验性精索静脉曲张大鼠生精细胞凋亡状况的影响

目的:探讨实验性精索静脉曲张大鼠睾丸生精细胞凋亡状况以及生精冲剂对其凋亡的影响。方法:将60只成年雄性W istar大鼠随机抽出20只为对照组,其余按石津和彦改良法制成左精索静脉曲张大鼠模型,再随机分为模型组20只,治疗组20只。用末端脱氧核苷酸转移酶介导的原位缺口末端标记法(TUNEL)检测睾丸生精细胞凋亡。结果:模型组大鼠睾丸生精细胞凋亡指数明显高于对照组(P<0.01)和治疗组(P<0.01),治疗组与对照组比较有明显差异(P<0.01)。结论:实验性精索静脉曲张大鼠睾丸生精细胞凋亡增加,这可能是影响生育力的机制之一;生精冲剂能够减少精索静脉曲张大鼠睾丸生精细胞凋亡,对睾丸生精功能具有保护作用,进而可提高睾丸的生殖能力。     

腹股沟区皮下埋藏睾丸对生精细胞凋亡及Bcl-2/Bax蛋白表达的影响

目的:通过观察腹股沟区皮下埋藏睾丸生精细胞的凋亡及Bcl-2/Bax蛋白表达,探讨二者的关系。方法:以健康育龄期36只雄性新西兰大白兔为实验动物,随机分成实验组18只、对照组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。动物模型建立后第8周末,每组随机取6只大白兔测量睾丸表面温度后进行睾丸活检。睾丸组织行原位末端标记(TUNEL)法检测生精细胞凋亡、免疫组化法结合图像分析仪检测睾丸生精细胞Bcl-2、Bax蛋白的表达。结果:对照组的睾丸表面温度为(36.15±0.64)℃,实验组为(38.02±0.36)℃,两组比较差异有统计学意义(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(89.69士3.76)%,对照组为(7.73±4.95)%,两者比较有显著差异(P<0.05)。实验组Bax表达显著升高(P<0.05),Bcl-2表达显著降低(P<0.05),凋亡细胞主要为初级精母细胞和圆形精子细胞。结论:腹股沟区皮下埋藏睾丸局部温度升高诱导睾丸的生精细胞凋亡增加,生精细胞凋亡增加与Bcl-2蛋白表达降低、Bax蛋白表达升高密切相关。Bcl-2/Bax的改变是高温引起生精细胞凋亡的机制之一。     

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。     

抗氧化酶对隐睾生精细胞凋亡的影响

目的 :探讨超氧化物歧化酶 (SOD)、过氧化氢酶 (CAT)、谷胱甘肽过氧化物酶 (GSHPx)对隐睾生精细胞凋亡的影响。　方法 :4 8只未成熟SD雄性大鼠随机分为隐睾组和对照组 ,于术后第 1、3、7d采集睾丸。TUNEL法检测其生精细胞凋亡 ;化学比色法测定其SOD、CAT、GSHPx的活性和丙二醛 (MDA)含量。　结果 :术后第 7d ,与对照组相比 ,隐睾重量、SOD活性、CAT活性和SOD/ (CAT +GSHPx)比值均显著降低 (P均 <0 .0 1) ,GSHPx的活性无显著变化 (P >0 .0 5 ) ,生精细胞凋亡指数和MDA含量均显著增加 (P均 <0 .0 1)。　结论 :隐睾生精细胞的凋亡与抗氧化酶活性的降低密切相关     

精索静脉曲张大鼠生精细胞凋亡与caspase-3蛋白的表达

目的:探讨caspase-3在精索静脉曲张大鼠生精细胞中的表达及其与生精细胞凋亡的关系。方法:雄性SD大鼠24只,随机分为假手术组(SOG)、30 d术后组(PG1)、60 d术后组(PG2),每组8只。建立左精索静脉曲张模型,TUNEL法检测生精细胞凋亡,免疫组化SABC法检测caspase-3表达。结果:SOG、PG1、PG2大鼠左、右侧睾丸每生精小管截面caspase-3阳性细胞数分别为0.117 5±0.012 9、0.246 3±0.042 1、0.293 8±0.051 1及0.165 0±0.019 2、0.253 8±0.021 9、0.277 5±0.034 3。与SOG相比,PG1和PG2组大鼠双侧睾丸生精细胞caspase-3表达均增加,差异有显著性(P=0.011 5及P=0.014 4)。结论:精索静脉曲张可能是大鼠生精细胞caspase-3表达增加且生精细胞凋亡增多的机制之一。     

糖皮质激素对大鼠生精细胞作用的研究

本文在建立糖皮质激素作用的大鼠的体内模型基础上,采用TUNEL染色,经激光共聚焦显微镜分析、透射电镜观察等方法,研究糖皮质激素对大鼠生精细胞凋亡的作用。结果表明,高浓度的糖皮质激素能显著增加大鼠生精细胞凋亡。     

大鼠隐睾睾丸复位固定术前后生精细胞凋亡与Bcl-2和Bax基因表达的研究

<正>隐睾是男性常见生殖系统畸形,可影响男性生育能力,是男性不育的重要原因之一。隐睾复位固定术是使隐睾患者恢复生育能力的有效途径。睾丸固定术能否完全恢复生精功能,何时行睾丸固定术对睾丸发育影响最小,生精功能恢复的指标如何判定等问题,迄今尚未见相关报道。     

茶多酚对精索静脉曲张大鼠睾丸生精细胞凋亡的影响

目的:研究茶多酚对精索静脉曲张大鼠睾丸生精细胞凋亡的影响,分析茶多酚对精索静脉曲张大鼠生精功能是否具有保护作用。方法:清洁级青春期Wistar大鼠32只随机分为4组,每组8只:假手术组(仅模拟手术过程,不结扎血管)、模型组(建立左精索静脉曲张模型)、茶多酚低剂量组[建立精索静脉曲张模型并给予10 mg/(kg·d)茶多酚干预]、茶多酚高剂量组[建立精索静脉曲张模型并给予40 mg/(kg·d)茶多酚干预]。建立左侧精索静脉曲张模型4周后,假手术组及模型组均给予生理盐水1 ml/100 g,茶多酚低剂量组及茶多酚高剂量组分别给予茶多酚10 mg/kg(用生理盐水配置成浓度为1 mg/ml)和40 mg/kg(用生理盐水配置成浓度为4 mg/ml),灌胃,1次/日,持续4周。4周后4组大鼠均处死并分别取左侧睾丸组织,检测低氧诱导因子-1(HIF-1)、Bcl-2、Bax、细胞色素C(Cyt C)和caspase-3在睾丸组织中的表达及生精细胞的凋亡并计算凋亡指数(AI)。结果:茶多酚干预组大鼠Bcl-2表达高于模型组(110.03±3.07),低于假手术组(154.18±2.96);茶多酚低剂量组大鼠Bcl-2(127.67±1.28)表达低于茶多酚高剂量组(136.41±1.95),差异有统计学意义(P0.05)。茶多酚干预组大鼠HIF-1、Bax、Cyt C和caspase-3的表达低于模型组,高于假手术组;茶多酚低剂量组大鼠HIF-1、Bax、Cyt C和caspase-3的表达高于茶多酚高剂量组,差异有统计学意义(P0.05)。假手术组AI最低,茶多酚干预组AI低于模型组,茶多酚高剂量组AI低于茶多酚低剂量组,差异有统计学意义(P0.05)。结论:茶多酚能显著降低精索静脉曲张大鼠睾丸生精细胞的凋亡。     

Effect of local testicular heating on spermatogenic cell apoptosis in rats

Aim: To study the effect of testicular local heating on spermatoge-nic cell apoptosis in rats. Methods: Seventy male SD rats were divided into the heat-treated and the control groups. The former was exposed to heat (43 ℃) for 12 hours. Each group was further divided into seven subgroups with respect to the time of observation after heat exposure, i.e., 12 h and 1 days, 3 days, 6 days, 10 days, 50 days and 80 days, respectively. In each subgroup, sper-matogenic cell apoptosis was examined by means of electron microscopy, flow cytometry and terminal deoxynucleotidyl trans-ferase-mediated dUDP-nick end labeling (TUNEL) methods. Results: The percentage of cells with sub-haploid and the percentage of positive TUNEL cells were significantly higher in the heat-treated groups than in the controls (P<0.01). The reaction of cell apoptosis to local heat was highly selective: spermatocytes were the most sensitive, followed by spermatids, spermatozoa and sper-matogonia in a decreasing order. Conclusion: Local testic     

青春期营养性肥胖大鼠睾丸生精细胞周期的变化

目的探讨青春期营养性肥胖大鼠的睾丸生精细胞周期的变化。方法80只新生的雄性清洁级SD大鼠随机分为两组,对照组(n=32只)喂普通饲料,肥胖组(n=48只)喂高脂饲料,分别于喂养后的第3、4、5、6周末观察喂养后体重,计算Lee’s指数,流式细胞分析术检测睾丸生精细胞周期的改变。结果与对照组比较,肥胖组大鼠第3周开始体重有显著性增加(P<0.05),6周内体重持续上升,至第6周末肥胖组大鼠体重超过对照组达26.6%(P<0.01),Lee’s指数随着大鼠肥胖程度的增加而逐渐增高;大鼠G0/G1期细胞在高脂饲料喂养第3周末增多(P<0.05)、肥胖组S期细胞显著下降(P<0.01),此时处于G2/M期细胞的百分数明显增多(P<0.05)。结论青春期时的肥胖可以引起睾丸生精细胞S期细胞百分数逐渐减少,出现G2期细胞阻滞,细胞有丝分裂延迟。     

Ureaplasma urealyticum infection and apoptosis of spermatogenic cells

Aim: To study the relationship between Ureaplasma urealyticum (UU) infection and apoptosis of human spermato-genic cells. Methods: Spermatogenic cells were observed under light microscope with Wright-Giemsa staining andby means of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP)-biotin nick-end labeling(TUNEL) technique. Results: Apoptotic rate of UU-infected males (15.5 % ± 6.8 % ) was significantly higherthan that of controls (5.2 % ± 2.3 % ). Conclusion: Apoptosis of spermatogenic cells can be caused by UU in-fection, which provides further evidence for UU-induced male infertility. (Asian J Androl 1999 Sep ; 1: 127 - 129)     

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。     

Brief maternal exposure of rats to the xenobiotics dibutyl phthalate or diethylstilbestrol alters adult-type Leydig cell development in male offspring

Maternal exposure to estrogenic xenobiotics or phthalates has been implicated in the distortion of early male reproductive development, referred to in humans as the testicular dysgenesis syndrome. It is not known, however, whether such early gestational and/or lactational exposure can influence the later adult-type Leydig cell phenotype. In this study, Sprague–Dawley rats were exposed to dibutyl phthalate (DBP; from gestational day (GD) 14.5 to postnatal day (PND) 6) or diethylstilbestrol (DES; from GD14.5 to GD16.5) during a short gestational/lactational window, and male offspring subsequently analysed for various postnatal testicular parameters. All offspring remained in good health throughout the study. Maternal xenobiotic treatment appeared to modify specific Leydig cell gene expression in male offspring, particularly during the dynamic phase of mid-puberty, with serum INSL3 concentrations showing that these compounds led to a faster attainment of peak values, and a modest acceleration of the pubertal trajectory. Part of this effect appeared to be due to a treatment-specific impact on Leydig cell proliferation during puberty for both xenobiotics. Taken together, these results support the notion that maternal exposure to certain xenobiotics can also influence the development of the adult-type Leydig cell population, possibly through an effect on the Leydig stem cell population.     

Effect of spinal cord injury on iNOS and Bcl-2 gene expression in spermatogenic cell of male rats

Effect of spinal cord injury on iNOS and Bcl-2 gene expression in spermatogenic cell of male rats     

Effect of prenatal exposure to diethylstilbestrol on gubernacular development in fetal male mice

AIM: To study the effect of prenatal exposure to diethylstilbestrol (DES) and the role of actin and proliferating cell nuclear antigen (PCNA) on testicular gubernaculum development in fetal male Kunming mice. METHODS: Pregnant mice were randomly assigned to 6 groups and injected with DES subcutaneously from gestational day 9 (E9) to day 17 (E17) at doses of 0, 25, 50, 100, 200 microg.kg-1.d-1 in 0.2 mL dimethyl sulfoxide (DMSO). On E17 they were sacrificed and fetuses quickly removed for fixation. Male fetuses were sliced on serial coronal plane. Histological changes were observed under the light microscope (LM) and ultrastructural changes with the scanning and transmission electron microscopes (SEM and TEM). The expression intensity of actin and PCNA in the gubernacula was quantitated by immunohistochemistry. RESULTS: The mortality of the fetuses was higher in the DES-treated groups than that in the DMSO and saline controls (P<0.05). Under LM the gubernacula were seen to be poorly developed with smaller bulbs. On SEM the bulbs lose the clear demarcation between the mesenchymal inner core and the muscular outer layer and looked like a small cone instead of the normal cylindrical appearance. On TEM there were some smaller disordered myofibrils and sparse cytoplasmic organelles in the gubernacular muscular cells of the treated groups. The expression intensity of actin and PCNA in the gubernacula was significantly weaker in the treated groups than that in the DMSO and saline controls (P<0.05). CONCLUSION: DES induces underdevelopment of the gubernacula in a dose-dependent manner in fetal male mice and down regulates the actin and PCNA expression.