锰对雄性大鼠睾丸毒性的病理学研究

每天给大鼠腹腔注射氯化锰１５ｍｇ／ｋｇ，连续６０天及８４天。结果染锰大鼠睾丸重量下降（Ｐ＜０．０１），睾丸脏器系数也明显降低（Ｐ＜０．０１）；血清锰虽无明显增高，但睾丸组织中的锰含量却增高；光镜检查；睾丸曲细精管呈不同程度的变性，管内生精细胞数目减少，精子形成少或无。     

新生期甲醛染毒对成年后大鼠雄性性行为、睾丸重量以及血清睾酮水平的影响

目的观察新生期甲醛染毒对成年后大鼠雄性性行为、睾丸重量和血清睾酮水平的影响。方法选用健康清洁级新生7日龄雄性SD大鼠24只,随机分为高剂量甲醛（10mg/m3）、低剂量甲醛（0.1mg/m3）染毒组和对照组3组,甲醛染毒14d后常规饲养4周至成年。然后分别观察成年后雄性大鼠的扑捉潜伏期（CLP）和60min内扑捉雌鼠的次数（CT）,称量睾丸重量以及利用放免法测定大鼠血清睾酮（T）水平。结果低剂量甲醛染毒组大鼠CLP,CT,睾丸重量以及血清睾酮含量与对照组相比没有明显差异。但高剂量甲醛染毒组大鼠的扑捉潜伏期（CLP）与对照组相比明显升高（P〈0.05）;60min内扑捉雌鼠的次数（CT）、睾丸重量与对照组相比均明显下降（P〈0.05）;大鼠血清睾酮水平与对照组和低剂量甲醛染毒组相比轻度下降。结论新生期甲醛染毒对成年后雄性大鼠性行为以及睾丸有一定的损伤作用,并且损伤具有剂量依赖性。     

亚慢性染毒丙烯酰胺对雄性大鼠生殖及睾丸内分泌功能的影响

目的:探讨亚慢性丙烯酰胺染毒对雄性大鼠生殖及睾丸内分泌功能的影响。方法:选择SD雄性成年大鼠40只,随机分成4组,每组10只,灌胃给予丙烯酰胺,剂量分别为0、4、10、18mg/(kg.d),染毒9周。染毒结束后,测量大鼠后肢支撑力,精子存活率、精子畸形率、睾丸匀浆中碱性磷酸酶(ALP)和酸性磷酸酶(ACP)活性、血清和睾丸匀浆中T及E2浓度。建立睾丸Leydig细胞体外原代培养模型,丙烯酰胺体外染毒剂量分为0、0.1、0.75、4、8mmol/L,通过CCK-8法观察Leydig细胞活性。结果:随着染毒剂量的增加,后肢展开距离显著加宽(P<0.01)。精子存活率分别为(76.86±5.46)%、(65.43±5.16)%、(60.86±4.26)%和(46.86±2.73)%,各剂量组与对照组比较显著下降(P<0.01);畸形率分别为(39.00±10.95)%、(35.43±7.54)%、(45.71±13.28)%和(56.71±17.01)%,10、18mg/(kg.d)剂量组明显上升(P<0.05)。ACP活性为(82.93±11.05)、(73.52±8.77)、(77.67±3.04)、(68.56±3.09)U/gprot,呈下降趋势;ALP活性为(0.96±0.15)、(1.07±0.22)、(1.12±0.22)、(0.74±0.10)U/gprot,呈现先上升后下降的趋势。两者的活性在18mg/(kg.d)剂量组与对照组比较差异显著(P<0.05)。血清T浓度分别为(13.44±4.76)、(7.69±3.84)、(5.23±1.42)、(1.36±0.86)ng/ml,睾丸匀浆中T浓度分别为(4.95±1.64)、(3.01±0.76)、(2.44±0.91)、(0.85±0.49)ng/mgprot,两者各剂量组与对照组比较都显著下降(P<0.01)。各剂量组E2水平无明显差异。丙烯酰胺染毒24h后,培养细胞A值分别为0.82±0.06、0.56±0.07、0.44±0.06、0.26±0.03和0.45±0.21,0.1、0.75、4、8mmol/L剂量组Leydig细胞活性受到显著抑制(P<0.01)。结论:亚慢性丙烯酰胺染毒,影响精子正常发育,引起睾丸一些生化酶活性改变;大鼠后肢运动协调性明显受到影响;丙烯酰胺对Leydig细胞有直接损伤作用,影响其内分泌功能。     

枸杞、黄芪对大鼠睾丸支持细胞功能的影响

目的:探讨中药枸杞、黄芪对大鼠睾丸支持细胞(Sertoli细胞)功能的作用及机制。方法:对18~22 d的SD大鼠睾丸Sertoli细胞进行分离培养,用MTT法检测枸杞(50、100、200、400μg/m l枸杞多糖)、黄芪(12.5、25、50、100 g/L黄芪提取液)对Sertoli细胞增殖的影响,用一步法RT-PCR方法检测枸杞、黄芪对正常培养状态和过氧化物(H2O2)损伤状态下Sertoli细胞抑制素(INH)βB亚基转录的影响。结果:高浓度枸杞(400μg/m l)、黄芪(100 g/L)对Sertoli细胞的增殖有促进作用(P<0.05),高浓度杞芪合剂(400μg/m l枸杞+100 g/L黄芪)对Sertoli细胞增殖有明显促进作用(P<0.01);在正常培养状态下,枸杞、黄芪及杞芪合剂对Sertoli细胞INHβB亚基的转录具有明显促进作用(P<0.01);在过氧化物(H2O2)损伤状态下,Sertoli细胞INHβB亚基的转录明显降低(P<0.01),黄芪对Sertoli细胞INHβB亚基的转录水平具有上调作用(P<0.05),枸杞、杞芪合剂对其转录水平具有明显上调作用(P<0.01)。结论:枸杞、黄芪及杞芪合剂对体外培养的Sertoli细胞INHβB亚基的转录具有促进和保护作用。     

尾悬吊状态对性成熟期雄性大鼠生殖功能的影响

目的:探讨尾悬吊造成的模拟失重状态对性成熟期雄性大鼠生殖功能的影响及其机制,为研究太空环境对人类生殖功能的影响打基础。方法:性成熟期健康SD雄性大鼠40只,随机分为4组,每组10只,分为实验1组(尾悬吊14 d)、实验2组(尾悬吊28 d)和对照1组(自由活动14 d)、对照2组(自由活动28 d),观察睾丸的重量和形态学改变、精子数量和质量改变、血液中激素含量的改变,并利用原位缺口末端标记法(TUNEL)检测睾丸细胞的凋亡。结果:各悬吊组大鼠睾丸重量与相应对照组相比明显下降(P<0.05),附睾精子数量和活动率明显减少(P<0.05),精子畸形率和凋亡率明显增加(P<0.05),卵泡刺激素和黄体生成素轻度增高,而睾酮含量明显下降(P<0.05)。但上述变化在悬吊14 d组和28 d组之间无明显差异(P>0.05)。另外,悬吊组睾丸组织生精小管萎缩,生精上皮细胞层数逐渐减少,管腔内的精子数明显减少,悬吊28 d组比悬吊14 d组改变更明显。结论:模拟失重对性成熟期雄性大鼠生殖功能有较明显的损害,这种损伤可能与引起睾丸生精细胞凋亡有关。抑制睾丸生精细胞凋亡也许可以防护失重状态下的生殖损害。     

大鼠睾丸支持细胞溶酶体的周期性变化

作者在透射电镜下观察了大鼠精子发生过程中支持细胞（Ｓｅｒｔｏｌｉ细胞）中溶酶体的结构及分布的改变。结果Ｓｅｒｔｏｌｉ细胞中溶酶体数目多，在细胞中所处的位置有周期性变化，细胞中全部胞质和在基底部胞质中溶酶体的截面积占胞质截面积的比例在生精第ＶＩＩ阶段最大，近腔部溶酶体截面积占胞质截面积在第ＩＩ、ＩＸ阶段最小；Ｓｅｒｔｏｌｉ细胞总胞质和在基底部单位胞质截面积的溶酶体数在第ＶＩＩ阶段最大，近腔部单位胞质截面积溶酶体数在顶体期后几阶段最大，第ＩＸ阶段最小。Ｓｅｒｔｏｌｉ细胞溶酶体的周期性变化是与生精细胞相互影响、相互作用，也是精子连续正常发生的前提条件之一。     

双酚A对大鼠睾丸支持细胞糖代谢的影响

目的:观察不同浓度双酚A(BPA)对原代培养大鼠睾丸支持细胞(SC)糖代谢及乳酸脱氢酶(LDH)表达的影响,探讨BPA致男性不育的机制。方法:采用两步酶消化法分离雄性Wistar大鼠睾丸SC,建立SC原代培养模型,免疫组化鉴定Fas L。传代后SC随机分为对照组和实验组(100 nmol/L、10μmol/L和1 mmol/L BPA)。培养48 h后,CCK-8法测定细胞增殖活性,核磁共振波谱法测定细胞内代谢物浓度,RT-PCR及Western印迹检测LDH的表达。结果:体外分离培养SC的纯度为(96.05±1.28)%(n=10),CCK-8实验结果显示:与对照组相比,暴露于100 nmol/L BPA组[(98±8)%]、10μmol/L BPA组[(96±3)%]和1 mmol/L BPA组[(95±3)%]的细胞增殖率无明显变化(P0.05);核磁共振波谱显示:与对照组相比,10μmol/L和1 mmol/L BPA作用下SC内葡萄糖及乳酸浓度明显降低(P0.05);RT-PCR及Western印迹结果显示:LDH mRNA的表达随BPA浓度升高呈现降低趋势(100 nmol/L、10μmol/L及1 mmol/L组P均0.05),而LDH蛋白的表达只在1 mmol/L组明显降低(P0.05)。结论:较高浓度BPA降低LDH,影响SC糖代谢过程。推测BPA通过影响SC糖代谢,减少向生殖细胞乳酸的供给,影响精子发生过程。     

五氯酚对大鼠睾丸支持细胞影响的研究

目的:检测五氯酚(PCP)对体外培养的大鼠睾丸支持细胞的毒性作用。方法:35日龄雄性SD大鼠4只,采用MTT法检测浓度为10-2、10-1、1、10μmol/LPCP对大鼠睾丸支持细胞活力的影响以及AnnexinV/PI流式细胞术检测PCP对大鼠支持细胞的毒性作用。结果:支持细胞的活力随PCP浓度的提高而明显降低并呈时间依赖效应;随浓度增高,细胞由胞质丰富、胞核饱满变为胞质稀疏、胞核皱缩、细胞数目减少,进而细胞界限不明显乃至消失;流式细胞仪检测结果表明,PCP导致坏死的支持细胞比例增加。结论:PCP可引起大鼠睾丸支持细胞坏死,细胞毒性作用明显。     

脂多糖对雄性大鼠睾丸组织病理学和生殖内分泌功能的影响

目的:研究脂多糖诱导的炎症对雄性大鼠睾丸组织病理学和生殖内分泌功能的影响,初步探讨炎症影响雄性生育的可能机制。方法:36只雄性SD大鼠(400~450g)随机分为4组,每组9只。对照组(A组)大鼠腹膜内注射无菌生理盐水,余下3组大鼠腹膜内注射脂多糖(LPS,5 mg/kg,溶于无菌生理盐水中),分别于注药12 h(B组)、24 h(C组)和72 h(D组)后麻醉处死。取各组大鼠左侧睾丸,制成组织切片,HE染色,镜下观察睾丸组织病理学改变;取各组大鼠血清进行血清睾酮(T)、卵泡刺激素(FSH)和黄体生成素(LH)含量检测。结果:①大鼠睾丸组织HE染色显示:与A组相比,B组大鼠睾丸生精小管结构清晰,各级生精细胞排列整齐,部分生精小管管腔内精子数目有所减少,并可见脱落的生精细胞;C组大鼠睾丸生精上皮变薄,生精小管结构较紊乱、不规则,各级生精细胞减少且排列不整齐,生精小管管腔内成熟精子数目减少,并可见脱落的生精细胞;D组大鼠睾丸生精上皮变薄,生精小管结构紊乱、不规则,各级生精细胞明显减少且层次紊乱,生精小管管腔内成熟精子数目减少,可见较多生精细胞脱落并阻塞管腔。②各组大鼠血清T、LH、FSH含量,A组T为(0.490±0.028)ng/ml,LH为(6.290±0.515)ng/L,FSH为(1.837±0.127)IU/L;B组T为(0.460±0.024)ng/ml,LH为(5.881±0.124)ng/L,FSH为(1.707±0.098)IU/L;C组T为(0.417±0.021)ng/ml,LH为(5.123±0.271)ng/L,FSH为(1.620±0.115)IU/L;D组T为(0.378±0.021)ng/ml,LH为(4.504±0.279)ng/L,FSH为(1.562±0.216)IU/L;与A组相比,B组大鼠血清T、LH和FSH含量均降低,但无统计学差异(P0.05);C、D组大鼠血清T和LH明显降低(P0.01),FSH降低,有统计学差异(P0.05)。结论:脂多糖(5 mg/kg)腹膜内注射诱导炎症可损害雄性大鼠睾丸组织,并影响大鼠生殖内分泌功能,导致血清T、LH和FSH含量降低,可能影响雄性生育。     

An assessment of inhibin-like activity secreted by Sertoli cells in culture using castrated adult male rats

Sertoli cells isolated from testes of 16–18 days old male rats were maintained in culture. Incubation media from these culture were pooled on day 7 and tested for its inhibin-like activity either with (SCCM) or after charcoal treatment (CSCCM) in castrated adult male rats. The assay was based on the tacit assumption that SCCM or CSCCM would specifically lower circulating blood serum levels of FSH. Subcutaneous (sc)injections of CSCCM at a dose level of 1 mg protein per rat, per day, x 3 days caused a specific suppression of FSH levels, while lower dosages of CSCCM (Protein content of 300 μg or 600 μg/rat/day, x 3 days) were without any affect on basal levels of FSH and LH. SCCM was ineffective at all dose levels tested. Intracardiac injections of varying doses of LHRH (25 to 400 ng/rat) to CSCCM pre-treated rats (200 μg/rat/day, x 3 days) failed to increase the levels of LH and FSH. These results support the presence of inhibin like activity in SCCM by a bioassay procedure alternate to in vitro pituitary cell culture system used by other investigators.     

Ethanol‐induced mitophagy in rat Sertoli cells: Implications for male fertility

Autophagy is a pro‐survival mechanism involving lysosomal degradation of damaged cellular components following multiple forms of cellular stress. There is currently a lack of literature on the mechanism, and specifically on mitophagy (selective autophagy of damaged pro‐apoptotic mitochondria) in Sertoli cells (SCs). Against such a background, the authors induced mitophagy in SCs of adult male rats using a single injection of ethanol (5 g/kg) and observed mitophagy in the SCs via transmission electron microscopy 24 hr later. In addition, we briefly discussed the possible clinical implications of enhanced autophagy and mitophagy in stressed SCs in our model and in other models of acute stress (e.g., heat and transplantation stress). Further studies on SC autophagy are required, as a full understanding of the molecular mechanisms controlling autophagy in stressed SCs may have therapeutic implications for infertility treatment.     

不同途径行Sertoli细胞-肝脏联合移植对肝移植术后急性排斥的影响

目的通过不同途径行Sertoli细胞-肝脏联合移植，探讨Sertoli细胞是否可为移植肝提供免疫保护。方法“2步法”分离培养Sertoli细胞，“二袖套管法”行大鼠原位肝移植并以Wistar→SD组合建立排斥反应模型。通过三种途径进行Sertoli细胞-肝脏联合移植。分别观察术后各组症状、体征、肝功能变化、移植肝病理特征等。采用免疫组化、凋亡等技术检测Sertoli细胞功能及作用，探讨其对肝移植急性排斥的影响。结果肝移植急排模型不干预组14只，1只存活超过14d。Sertoli细胞腹腔注射组、阴茎背静脉注射组和供体移植前门静脉注射组分别有5、8、7只存活超过14d。各干预组存活率与对照组比较：后两组差异有显著性（P〈0．05），腹腔注射组差异不显著（P〉0．05）；各干预组之间存活率差异无显著性（P〉0．05）。供肝病理检查显示各干预组排斥反应较对照组轻。免疫组化及凋亡检测发现：肝移植后14d，Sertoli细胞仍存活并表达FasL，Sertoli细胞周围有淋巴细胞集聚及凋亡的淋巴细胞。结论Sertoli细胞对肝移植急性排斥有抑制作用，对供肝有诱导免疫耐受作用，Sertoli细胞通过Fas／FasL途径诱导淋巴细胞凋亡。     

青春期己烯雌酚暴露对大鼠睾丸发育及功能的影响

目的 探讨青春期己烯雌酚(diethylstilbestrol,DES)暴露对SD(Sprague-Dawley)大鼠睾丸发育及功能的影响.方法 35日龄雄性SD大鼠90只,随机分为DES 0.01、0.1、1.0、10.0 μg·kg-1·d-14个实验组和1个对照组(编码为Bda、BDb、BDc、BDd和BC组,每组18只).于青春期,即出生后第36天(postnatal day 36,PND 36)至PND 49,实验组每日皮下注射相应剂量的DES,共14 d,对照组仅注射溶媒.于青春期晚期(PND 50)、性成熟后(PND 64)和成年期(PND 130)分3批(每批6只)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PND 130大鼠附睾尾精子质量.结果 PND 50时,BC、Bda、BDb、BDc和BDd组单侧睾丸重量分别为(1.26±0.13)、(1.23±0.20)、(0.99±0.15)、(0.85±0.23)和(0.60±0.04)g,其中BDb、BDc和BDd组均较BC组减轻(P＜0.05);与BC组比较,BDb组仅有少数生精小管生精上皮中的细胞数目稍减少,BDc和BDd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.PND 64时,BC、Bda、BDb、BDc和BDd组单侧睾丸重量分别为(1.54±0.14)、(1.55±0.17)、(1.52±0.11)、(1.37±0.14)和(0.88±0.15)g,其中BDc和BDd组均较BC组减轻(P＜0.05);BDc和BDd组睾丸组织形态学改变与PND 50时类似,但较PND 50时有所改善.PND 130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未见明显差异;BC、Bda、BDb、BDc和BDd组大鼠附睾尾精子密度分别为(71.00±14.85)、(69.00±23.98)、(67.00±13.52)、(31.67±12.94)和(18.83±6.68)×106/ml,其中BDc和BDd组精子密度均较BC组明显降低(P＜0.01);与BC组比较,BDd组精子活动率下降(P＜0.01),BDb、BDc和BDd组A级精子比例降低(P＜0.05),BDd组B级精子比例降低(P＜0.01).结论 青春期小剂量DES(0.01μg·kg-1·d-1×14 d)暴露对SD大鼠睾丸发育及功能无明显影响,大剂量DES(1.0～10.0 μg·kg-1·d-1×14 d)暴露对大鼠睾丸发育及功能具有明显的近期(PND 50和PND 64)和较远期(PND 130)毒性作用,该毒性作用随DES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密切相关.     

Enzymatic properties of adult human Sertoli cells in vitro

Summary. The adult human Sertoli cells produced lactate, estradiol-17β, transferrin and inhibin; germ cells modulate synthesis of these compounds. In order to study the functional features of human Sertoli cells in vitro , the aim of this study was to measure the lactic dehydrogenase (LDH), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP) and creatine kinase activities (CK) in primary cultures of Sertoli cells prepared from young men (mean age: 29 years, n = 11). Five LDH isozymes have been found in Sertoli cells, the main fractions being the LDH3 and LDH4; each of them represented 30% of the total LDH activity. Furthermore, CK and ALP activities were measured in Sertoli cells. It is of note that the Sertoli cell ALP activity was 50% lower than that of germ cells. Whatever the Sertoli cell parameter measured herein, there is a great variability between patients and FSH (dbc AMP, as well as retinol, insulin, testosterone) is poorly effective in improving these enzymatic activities in vitro. We have confirmed that GGT was exclusively present in Sertoli cells and thus may be considered as a specific marker. LDH is involved in Sertoli cell glucose transformation and thus provided energetic substrates for germ cells. In contrast, the roles of CK and ALP remains to be clarified. In conclusion, we have demonstrated the existence of several enzymes namely LDH, GGT, ALP and CK in Sertoli cells prepared from adult human testis.     

胰岛素替代治疗对青春期糖尿病鼠睾丸间质和支持细胞功能的影响

以４０日龄雄性Ｗｉｓｔａｒ大鼠４０只，分为对照（Ｃ），糖尿病（Ｄ），糖尿病胰岛素及时治疗（ＤＣＩＲ）和糖尿病胰岛素延迟治疗（ＤＤＩＲ）组，进行睾丸间质细胞和支持细胞功能的研究。结果表明：血清睾酮Ｄ组最低，培养间质细胞睾酮分泌量ＤＣＩＲ、ＤＤＩＲ和Ｄ组均显著低于Ｃ组（Ｐ＜０．０５及０．００１），ｃＡＭＰ分泌量ＤＤＩＲ和Ｄ组显著低于Ｃ和Ｄ分别的ＣＩＲ组（Ｐ＜０．０１）；睾丸组织雄激素结合蛋白（ＡＢＰ）水平，ＤＤＩＲ组显著高于Ｃ和Ｄ组（分别为Ｐ＜０．０５及Ｐ＜０．０１），其它组间比较差异不显著，培养支持细胞ＡＢＰ分泌量Ｄ组显著低于Ｃ和ＤＣＩＲ组（Ｐ＜０．０５），ｃＡＭＰ分泌量ＤＤＩＲ和Ｄ组显著低于Ｃ和ＤＣＩＲ组（Ｐ＜０．０１）。     

Effects of exposure to a mobile phone on testicular function and structure in adult rabbit

The accumulating effects of exposure to electromagnetic radiation emitted by a conventional mobile phone (standby position) on the testicular function and structure are not yet fully investigated. To study these effects longitudinally, a total of 24 adult male rabbits were randomly and equally divided into three groups. Rabbits in the first (phone) group were exposed, in specially designed cages, to radio frequency emitted from the mobile phone (800 MHz) in a standby position opposite to that of testes for 8 h daily for 12 weeks. The second group consisted of the stress controls which were kept in the same kind of cages to appreciate any cage-induced anxiety. The third group included the ordinary controls which were kept in the conventional roomy cages. Semen analysis and sperm function tests (viability, hypo-osmotic swelling and acridine orange) were conducted weekly. Histological testicular sections and serum total testosterone were also evaluated. A drop in the sperm concentration appeared in the phone group at week 6. This became statistically significant at week 8, compared with the two control (stress and ordinary) groups (133, 339 and 356 × 106/mL, respectively) and to the initial sperm count (341 × 106/mL) of this group. Motile sperm population showed similarity amongst the three study groups until week 10 when it declined significantly, and thereafter in the phone and stress control groups, with more significant decline in the phone animals (50, 61 and 72.4%, respectively). Histological examination showed also a significant decrease in the diameter of seminiferous tubules in the phone group vs. the stress and ordinary controls (191 μm vs. 206 and 226 μm, respectively). The other study points did not show any difference. In conclusion, low intensity pulsed radio frequency emitted by a conventional mobile phone kept in the standby position could affect the testicular function and structure in the adult rabbit.     

Reproductive function in male rats after brief in utero exposure to diethylstilboestrol

Long-term effects of brief in utero exposure to diethylstilboestrol (DES) during a foetal period known to be critical for gonadal development were evaluated. Rats were exposed to DES (100 microg/kg body-weight) from day 17 to 19 of pregnancy. All of the DES-treated pregnant rats (11/11) ate parts or whole of their offspring during the first day after birth (p=0.03). Surviving male offspring were examined on day 63 post-partum. DES induced a reduction in weight of the testis (p=0.06) and ventral prostate (p=0.07), even after this short exposure. DES tended to reduce the number of Sertoli cells (p=0.13). Our findings indicate that even a short in utero exposure of rats to DES during a critical period for gonadal development results in cannibalism and reduced testis and ventral prostate weight.     

己烯雌酚摄入对大鼠生精细胞凋亡及Bax和Bcl-2蛋白表达的影响

目的研究青春期己烯雌酚(diethylstilbestrol,DES)摄入对SD(Sprague-Dawley)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35d龄雄性SD大鼠,随机分为DES 0.01、0.1、1.0、10.0μg/kg·d~(-1)4个实验组和1个对照组(编码为BDa、BDb、BDc、BDd和BC组,每组n=6)。于青春期[出生后第36天(postnatal day 36,PND 36)至49d(PND 49)],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Bcl-2和Bax在生精细胞中的表达。结果与对照组相比,BDa组大鼠性成熟后生精细胞凋亡无明显变化,BDb、BDc和BDd 3组生精细胞凋亡增加,且随DES摄入剂量增加而有增加趋势。BC、BDa组生精细胞Bax相对弱表达而Bcl-2强表达,伴随DES摄入剂量增加,Bax表达逐渐增强而Bcl-2表达逐渐减弱,BDd组Bax强表达而Bcl-2弱表达。结论青春期较大剂量DES摄入可使大鼠性成熟后睾丸生精细胞凋亡增加,且随DES摄入剂量增加而有加强趋势。凋亡相关蛋白Bax和Bcl-2参与青春期DES摄入所致的生精细胞凋亡过程。