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。     

淫羊藿苷对酒精致雄性小鼠生殖损伤的影响

目的探讨淫羊藿苷对酒精致雄性小鼠生殖损伤的保护作用,为临床治疗酒精致男性不育提供理论依据。方法成年雄性小鼠随机分为正常组(蒸馏水)、模型组(酒精组)和给药组(酒精+淫羊藿苷组),灌胃5周后处死并制备附睾精子悬液,分别测定精子密度、精子活动率、存活率、精子畸形率;JC-1染色法检测线粒体膜电位改变;制备睾丸组织切片,观察睾丸组织病理改变。结果模型组小鼠精子密度、精子活动率和存活率明显低于正常组,精子畸形率明显增加;给药组精子密度、精子活动率和存活率较模型组明显升高,精子畸形率明显降低。线粒体膜电位分析,模型组去极化细胞(凋亡细胞)比例明显高于正常组;给药组去极化细胞(凋亡细胞)比例低于模型组。睾丸组织切片观察发现,与正常组比较,模型组生精细胞层数减少,结构紊乱稀疏;给药组生精小管结构明显改善。结论淫羊藿苷对酒精致雄性小鼠生殖损伤有一定保护作用。     

育精阴对雄性豚鼠免疫性不育的实验研究

目的　研究中药育精阴治疗免疫性不育的作用及其机理。方法　运用主动免疫法造成豚鼠实验性变态反应性睾丸炎 (EAO)模型 ,观察睾丸生精细胞和附睾尾部精子质量的变化。运用不同剂量育精阴灌胃 ,观察育精阴对生精上皮和附睾尾部精子质量的作用。结果　发现实验性变态反应性睾丸炎造成睾丸生精细胞退行性病变 ,附睾精子质量下降。育精阴可减轻和修复实验性变态反应对睾丸附睾的损伤 ,提高附睾精子质量。结论　育精阴对实验性变态反应性睾丸炎造成的免疫损伤有修复作用     

水通道蛋白抑制剂对大鼠精子成熟和生育影响的研究

目的通过水通道蛋白抑制剂对大鼠附睾精子成熟的影响，探索睾丸后抗生育作用靶点。方法不同剂量乙酰唑胺大鼠灌胃给药，连续10d。结束时将受试雄鼠与雌鼠合笼，计算交配指数、受孕指数、生育指数。采用计算机辅助分析系统分析精子活力形态。使用酶联免疫吸附法测定大鼠血清睾酮（T）和双氢睾酮（DHT）的浓度。睾丸附睾称重后进行HE染色和电镜分析。结果乙酰唑胺各剂量组大鼠精子活力明显降低，同时畸形率显著增加，导致雌鼠受孕率下降，而T、DHT和交配指数均无明显变化，睾丸和附睾亦无明显病理学改变。结论水通道蛋白抑制剂乙酰唑胺可以通过影响大鼠附睾精子成熟来抑制雄鼠生育，同时不影响睾丸和生殖内分泌系统。     

不同剂量昆明山海棠对雄大鼠生殖系统的形态学研究

雄性SD大鼠60只,随机分成6组,二组对照,其余4组用昆明山海棠稀醇提取物(ATH)灌胃给药,每日分别为2.0g/kg;1.0g/kg;及0.3g/kg(二组),每周六次。用药6周至完全不育后剖杀用药的前三组及一组对照。取睾丸、附睾、心、肝、脾、肺、肾、胃,行常规HE及PAS+苏木素染色,部分鼠做DNA、RNA组化反应。取附睾尾评价精子数据。0.3g/kg组及另一组对照继续以每日0.2g/kg的维持量灌胃16周以保持不育,停药5周后证实其生育力恢复后剖杀,观察指标同前。结果表明:2.0g/kg组曲细精管平均损伤率为55.4%,1.0g/kg组曲细精管损伤明显少于前组(损伤率6.2％),小剂量鼠无论给药6周或22周睾丸和附睾形态与对照无异。结果提示:大鼠睾丸曲细精管的损伤范围和损伤的程度与ATH的剂量大小有关,而与作用时间的长短关系不大。最低抗生育剂量时,对ATH最敏感的部位是变态期精子细胞及附睾精子,以此剂量长期给药对睾丸附睾无明显影响,所致不育可以恢复。     

直链烷基苯磺酸钠对雄性小鼠生精功能亚慢性毒性作用研究

目的 探讨直链烷基苯磺酸钠(LAS)对雄性小鼠生精功能的亚慢性毒性作用及毒性遗留作用. 方法 NIH种雄性小鼠给予LAS(630 mg/kg,315 mg/kg)灌胃,每日1次,连续经口染毒2个月,同时设对照组.在第4、8周及停止染毒后4周进行精子形态学观察,光镜观察睾丸组织形态学变化,透射电镜观察睾丸超微结构变化. 结果 实验组与对照组相比,染毒4周后精子畸形率上升,畸形主要表现在颈部,精子中部胞浆小滴增多.染毒8周精子畸形率升高更显著.组织学检查实验组小鼠染毒4周时精细小管内各级精母细胞和精子细胞数减少,生精细胞排列紊乱,随实验时间延长及染毒浓度的增加此异常改变更显著;超微结构观察显示睾丸生精上皮内支持细胞、间质细胞、精原细胞线粒体广泛异常改变,呈时间及浓度依赖性.上述异常改变在停止染毒后4周无明显恢复. 结论 LAS对雄性小鼠生精功能亚慢性毒性作用及毒性遗留作用表现为:使生精上皮内支持细胞、间质细胞及精原细胞出现广泛线粒体空泡样变,精子胞浆小滴出现率升高.     

氯化镉对雄性小鼠的生殖毒性作用

目的研究氯化镉对雄性小鼠生殖器官和生殖细胞的毒性作用以及对雄性小鼠生殖功能的影响。方法分别以每公斤体重0..5mg/kg、2mg/kg、8mg/kg体重腹腔染毒4周龄雄性小鼠,共10次。第50天以雌雄2:1同笼交配,观察雌鼠受孕率、生育子胎数、胎鼠重量。同时观察染毒雄鼠第50天时睾丸发育和睾丸指数、附睾精子数量、精子活动率和精子畸形率以及生殖细胞减数分裂。结果2mg/kg和8mg/kg组染毒小鼠睾丸发育受到影响,睾丸指数显著低于对照组和0.5mg/kg组(p<0.05,p<0.001)。2mg/kg组中部分小鼠(6/11)和8mg/kg组存活小鼠睾丸发育不良,无精子,不育。2mg/kg组小鼠1周受孕率和3周受孕率显著低于对照组和0.5mg/kg组(p<0.05),但染毒组异常妊娠率与对照组比差异无显著性。0.5mg/kg组雄鼠生育力、附睾精子数量、精子活动率和精子畸形率与对照组比较差异无显著性。结论2mg/kg浓度和更高浓度氯化镉致4周龄雄鼠睾丸发育不良,导致无精子症和不育,是造成生殖力降低的主要原因。     

吸烟对雄性小鼠睾丸间质细胞、睾酮及精子数量的影响

目的探讨吸烟对雄性小鼠的睾丸间质细胞、血清睾酮含量及附睾精子数量的影响。方法雄性小鼠50只,随机分为5组,即经吸烟处理后6周、12周(吸烟组),未经特殊处理的6周、12周小鼠(对照组),及吸烟6周后戒烟6周(戒烟组)小鼠。对5组小鼠睾丸组织进行HE染色,光镜下观察支持细胞、问质细胞形态及结构;免疫组化方法分别计数5组小鼠睾丸支持细胞及问质细胞数量;利用酶联免疫法(ELIsA)测定小鼠体内睾酮含量:用石墨炉原子吸收光谱法测定血清镉含量;同时计数附睾精子数。结果 HE染色可见吸烟6周小鼠睾丸问质细胞数量减少;吸烟12周小鼠睾丸问质细胞数量稀少甚至消失:而睾丸支持细胞数量及形态与对照组相比未见明显改变。免疫组化发现吸烟组小鼠睾丸支持细胞数量与对照组相比无显著差异(P0.05)。问质细胞数量随吸烟时间延长而显著减少(P0.01);小鼠血清中睾酮水平吸烟组明显低于对照组(P0.01),且随吸烟时间的延长而下降(P0.01):吸烟组小鼠附睾精子汁数较对照组显著下降(P0.01),并且与吸烟时间呈正相关(P0.01)。而戒烟组中,睾丸问质细胞、血清睾酮含量、附睾精子数量3组数据都较吸烟组增高。结论吸烟可导致睾丸间质细胞的破坏,影响体内睾酮的生成,进而导致生精障碍,而戒烟可逆转此现象。     

自身免疫性睾丸炎对精子特异性酶和生育功能的影响

目的 :研究自身免疫性睾丸炎对精子特异性酶和生育功能的影响。　方法 :复制豚鼠实验性变态反应性睾丸炎 (EAO)模型 ,采用酶动力学分光光度法和明胶固定底物薄膜法 ,观察EAO状态下精子顶体蛋白酶、透明质酸酶、精子胞质乳酸脱氢酶、附睾尾部精子和睾丸组织形态的变化。　结果 :EAO造成附睾精子顶体酶系中顶体蛋白酶、透明质酸酶和乳酸脱氢酶活性下降、附睾尾部精子质量下降、睾丸生精细胞发生退行性病变。　结论 :EAO明显影响雄性豚鼠生育力 ,睾丸生精细胞和附睾精子可能是主要作用环节。     

抗精子抗体对小鼠睾丸超微结构的影响

目的研究免疫性不育雄性小鼠的睾丸超微结构的特点,探讨抗精子抗体（AsAb）对睾丸生精微环境的影响。方法用同种精子及福氏佐剂免疫昆明种雄性小鼠。ELISA法检测AsAb,以此验证获得免疫性不育动物模型,同时设立对照组;解剖小鼠取其精子观察精子质量差别,透射电镜观察各组睾丸的超微结构。结果人工免疫后的雄鼠血清AsAb均为阳性。正常对照组均为阴性;模型组小鼠精子质量与对照组比较显著下降,生精上皮超微结构受损严重。结论用同种精子及福氏佐剂免疫动物,可成功制作抗精子抗体介导的免疫性不育动物模型;AsAb可通过攻击睾丸生精微环境影响雄性小鼠的生育力。     

鹿藿根4种提取物抗雄性小鼠生育作用比较

目的:比较4种鹿藿根提取物(乙醇提取物、乙酸乙酯提取物、正丁醇提取物及水溶物)的抗生育作用。方法:60只雄性昆明种小鼠分为生理盐水对照组、雷公藤多苷对照组、乙醇提取物组、乙酸乙酯提取物组、正丁醇提取物组及水溶物组,每组10只。各组药物浓度均为1%,雷公藤多苷为0.1%水溶液。每次0.1ml/10g,灌胃,1次/d,连续给药,共11周。在用药10周后,一对一雌雄合笼交配1周。分笼10d后处死雌鼠,观察妊娠率、活胎数、死胎数。11周后处死雄性小鼠观察附睾精子、附睾及睾丸病理变化情况及血清睾酮含量测定。结果:连续用药11周,与生理盐水对照组相比,实验组小鼠妊娠率下降,其中水溶物组最为明显(P<0.05)。雷公藤多苷对照组和水溶组精子明显减少(P<0.05)。组织学观察鹿藿根提取物对精原细胞、初级精母细胞影响不显著。各提取物组血清睾酮差异无显著性(P>0.05)。生理盐水对照组、雷公藤多苷对照组、乙酸乙酯提取物组、正丁醇提取物组、乙醇提取物组及水溶物组的抗精子发生效应积分分别0、(5.3±1.5)、(2.6±1.7)、(2.9±1.2)、(2.2±0.9)、(2.1±1.0)分。结论:4种鹿藿根提取物均有抗生育作用,但水溶物作用更强且对睾丸组织及睾丸生精能力影响较小。     

Antifertility effect of ethanolic extract of Amalakyadi churna in male albino mice

Aim: To evaluate the antifertility activity of the ethanolic extract of Amalakyadi churna by oral administra-tion in male albino mice. Methods: The ethanol extract of Amalakyadi churna at the dose of 250 mg/kg and 400 mg/kg body weight was administered orally for 30 days to adult male mice. On day 31, the mice were sacrificed and the testis and accessory reproductive organs were removed and weighed. The organs were processed for biochemical estimation and histological work. Results: Treatment with Amalakyadi chuma resulted in decrease in the weights of testis and accessory reproductive organs. The diameters of testis, seminiferous tubules and Leydig cell nucleus were decreased. The spermatogenic elements, like spermatogonia, spermatocytes and spermatids in the testis were re-duced significantly as well as the sperm count in cauda epididymis. There was a significant reduction in the protein,glycogen, DNA and RNA contents and the activity of acid phosphatase in the testis of extract treated mice compared with the control. The cholesterol content and the alkaline phophatase activity were increased significantly in treated mice. Conclusion: Amalakyadi churna extract arrests spermatogenesis in male mice without noticeable side effects.(Asian J Andro12003 Sep; 5: 247-250)     

长期给予睾酮致大鼠生精小管伴有生精细胞排列疏松

目的:确定睾酮致大鼠睾丸内睾酮抑制因而精子发生障碍是否伴有生精细胞排列疏松。方法:成年雄性SD大鼠肌注十一酸睾酮[19 mg/(kg.15 d)]130 d后取睾丸组织块,作甲基丙烯酸树脂包埋切片,观察睾丸的组织学变化。结果:除精子形成、精子释放障碍等改变以外,11.5%的生精小管轮廓内生精细胞的排列明显较疏松,成串、成束或成团的生精细胞(主要是精母细胞和精子细胞)之间出现朝向小管腔走行的放射状裂隙。结论:生精细胞排列疏松是大鼠睾丸内睾酮抑制所致重要组织学改变之一。     

唯支持细胞综合征患者的睾丸病理和性激素分析

目的 分析唯支持细胞综合征不育患者曲细精管的不同病理改变及其血清性激素水平,探讨是否可以为睾丸活检取精子或生精细胞提供参考。方法 测定107例唯支持细胞综合征患者的睾丸容积和血清性激素水平,做睾丸活组织检查,按活检结果分为完全无生精细胞与有生精细胞两组,并比较两组间差异有无显著性意义。结果 96例唯支持细胞综合征患者完全无生精细胞,11例患者有生精细胞或精子,完全无生精细胞患者与有生精细胞患者睾丸容积及性激素水平无差异(P>0.05)。结论 唯支持细胞综合征不育患者少数有精子或生精细胞发生,但缺乏明确的性激素指标反映曲细精管是否有精子或生精细胞存在。     

A stereological study of the effects of experimental inguinal cryptorchidism and subsequent orchiopexy on spermatogenesis in adult monkeys

Our previous study demonstrated that experimental intra-abdominal cryptorchidism in adult rabbits for 13 weeks resulted in severe spermatogenic arrest: type A spermatogonia was the only germ cell type seen in the seminiferous epithelium and its number per testis was reduced by 84%. Seven weeks following orchiopexy, the type A spermatogonial number returned to the near-normal range in most animals and spermatogenesis partially recovered (Reproduction 2002, 124, 95-105). This study aimed to determine whether inguinal cryptorchidism would produce less-severe damage to spermatogenesis and whether subsequent orchiopexy would better restore spermatogenesis. Five normal adult male rhesus monkeys (Macaca mulatta) underwent bilateral artificial inguinal cryptorchidism. Half a year later, one testis together with the ipsilateral epididymis were removed from each animal and then unilateral orchiopexy was performed on the contralateral side, with the remaining testis and epididymis being removed another half a year later. A contemporary unbiased and efficient stereological tool, the optical disector, was used to estimate numbers of all types of spermatogenic cells in the testis and spermatozoa in the epididymis. Spermatogenic arrest was induced by cryptorchidism at the stage of spermatogonia (n = 1), spermatocytes (n = 2) or early spermatids (n = 1), with the type A spermatogonial numbers per testis being reduced to 14.8-57.2% of the control average; in one of the five cryptorchid animals, however, spermatogenesis remained normal. Subsequent orchiopexy, which was successfully performed on two animals with cryptorchidism-induced spermatogenic arrest, brought on a full or partial recovery of spermatogenesis. In conclusion, inguinal cryptorchidism induces less severe (in comparison with an intra-abdominal one) and variable damage to spermatogenesis, which is restored, at least in part, by subsequent orchiopexy.     

睾丸细胞生物学研究进展

睾丸是男性生殖腺,由生精小管和间质构成。生精小管主要由生精细胞和支持细胞组成,是精子发生场所;间质中主要是间质细胞,间质细胞合成与分泌雄激素。本文介绍睾丸3种细胞的发育分化,以及成年期睾丸细胞的结构和生物学研究进展。     

二甲磺酸乙烷致间质细胞破坏所引起的成年大鼠睾丸和附睾的组织学变化:形态定量研究

目的:定量研究睾酮分泌剧烈减少所致睾丸和附睾的组织学变化。方法:14只成年 SD 大鼠腹腔内注射二甲磺酸乙烷(EDS,75mg/kg),14只注射生理盐水作为对照。7天后处死各组中的一半动物,过5天后处死另一半。取睾丸和附睾组织块,甲基丙烯酸树脂包埋。用体视学的光学体视框技术估计睾丸内的细胞数,并用其它形态定量研究方法获取另外一些参数。结果:EDS 注射使睾丸内的间质细胞几乎完全消失,但对支持细胞总数没有影响。EDS 注射7天后,生精上皮内可见许多长形精子细胞滞留,附睾管内可见许多圆形精子细胞。EDS 注射12天后,精子细胞和精母细胞的排列明显变疏松,生精细胞之间出现明显的裂隙,裂隙近似放射状朝向生精小管腔;睾丸内的非 B 型精原细胞总数和精母细胞总数与对照组相似,但 B 型精原细胞总数增加59%,而早期(圆形)、中期和晚期(长形)精子细胞总数分别减少37%、72%和52%。结论:EDS 所致精子发生损害主要是(1)精子释放障碍,(2)精子细胞、精母细胞分离并伴有精子形成和成熟分裂障碍。     

Histological changes of the testis and epididymis in adult rats as a result of Leydig cell destruction after ethane dimethane sulfonate treatment： a morphometric study

Aim： To quantitatively study the histological changes of the testis and epididymis as a result of a drastic reduction of testosterone secretion. Methods： Fourteen adult Sprague-Dawley rats were injected intraperitoneally with ethane dimethane sulfonate （EDS, 75 mg/kg） and the same number of animals were injected with normal saline as a control. At days 7 and 12 （after treatment）, respectively, half of the animals from each group were killed. The testes and epididymides were removed and tissue blocks embedded in methacrylate resin. The cell number per testis was estimated using the stereological optical disector and some other parameters were obtained using other morphometric methods. Results： The EDS treatment resulted in an almost complete elimination of Leydig cells but had no effect on the numbers of Sertoli cells per testis. At day 7 after EDS treatment, many elongated spermatids were retained in the seminiferous epithelium and many round spermatids could be seen in the epididymal ducts. At day 12, a looser arrangement of spermatids and spermatocytes became evident, with apparent narrow empty spaces being formed between germ cells in an approximately radial direction towards the tubule lumen; the numbers （per testis） of non-type B spermatogonia and spermatocytes were similar to controls, whereas that of type B spermatogonia increased by 59%, and that of early round, elongating and late elongated spermatids decreased by 37%, 72% and 52%, respectively. Conclusion： The primary spermatogenic lesions following EDS administration were （i） spermiation failure and （ii） detachment of spermatids and spermatocytes associated with impairment in spermiogenesis and meiosis.     

Ultrastructure of the seminiferous epithelium of ethyl methanesulphonate-treated mouse

Ethyl methanesulphonate (EMS) is a mutagenic alkylating agent that induces marked elevations of sperm abnormalities in mice. In this paper, we report the ultrastructural findings on the morphology of the seminiferous epithelium of mice resulting from EMS administration. Eight- to twelve-weeks-old male mice were injected intraperitoneally with EMS at 200 mg kg(-1) body weight daily for five consecutive days. Analysis of smears of epididymis and semi-thin sections of testes revealed that the more suitable specimens for the ultrastructural analysis were tissues of mice killed at the third week, following EMS administration. At this time, the spermatid was the damaged cell type. Abnormalities were mainly observed in the morphology of the nucleus, the acrosome, chromatin distribution and in the arrangement of the cytoplasmic microtubules, and binucleated spermatids were also observed. EMS has the capacity to penetrate the blood-testis barrier, and thus it can damage post-meiotic spermatogenic cells. However, morphological abnormalities could be the consequence of damage exerted on the differentiated spermatogonia stage, the most sensitive spermatogenic cell to the action of chemical agents or drugs. Our findings contribute to elucidate the action mechanism of the damage exerted by EMS administration on the germinal male cells.