棉酚对大鼠血睾屏障影响的研究

本实验采用辣根过氧化物酶、硝酸镧和氯化锂示踪观察成年大鼠灌服棉酚后血睾屏障的变化。成年雄性大鼠连续灌服醋酸棉酚30毫克/公斤/日2周、4周和6周后,血管内分别灌流0.75M 氯化锂溶液、2%硝酸镧溶液和睾丸内注射辣根过氧化物酶。结果表明,大鼠灌服棉酚达抗生育效果时,并未显示血睾屏障通透性损伤。因此,可以排除棉酚作用于支持细胞或血睾屏障继而间接影响生精细胞的可能性。     

棉酚对大鼠精母细胞~3H-精氨酸掺入作用的影响—淘析细胞分离术研究结果

本实验用大鼠灌服醋酸棉酚(每日30mg/kg体重)7、8、9和10周后,应用淘析器细胞分离术,获得了高纯度的粗线期精母细胞作~3H-精氨酸掺入能力的液闪测定,计算单位组蛋白重量中放射活性的脉冲数。结果表明,与对照组相比,棉酚组的粗线期精母细胞摄取精氨酸能力降低了35～50%。证明棉酚不仅对晚期精子细胞有影响,也使初级精母细胞的组蛋白合成代谢受到明显干扰。本文还讨论了有关的作用机理。     

棉酚的遗传学研究——Ⅰ.醋酸棉酚对大鼠雄性生殖细胞的细胞遗传学效应和血液中淋巴细胞微核测定的比较研究

本实验观察用醋酸棉酚短期灌服大鼠后雄性生殖细胞的细胞遗传学效应和血液中淋巴细胞微核的染色体损伤效应。给药组的精原细胞和精母细胞所出现的各类染色体畸变平均值虽比对照组高,但两组之间的差异没有统计学意义(P>0.05-0.10)。同样,给药组大鼠血液中淋巴细胞微核出现率也高于对照组,但也无统计学意义(P>0.05)。结果表明,醋酸棉酚对大鼠精原细胞和精母细胞的细胞遗传学效应与血液中淋巴细胞微核的染色体损伤效应的结果是一致的。     

服醋酸棉酚后幼龄大鼠睾丸间质细胞对hCG的反应性

本实验给15日龄大鼠每日灌服醋酸棉酚30mg/kg,共11天;同时从21天龄起每日皮下注射hCG 10IU,共5天后,观察了睾丸间质细胞的组织学及组织化学变化,并进行了定量及半定量测定。结果表明,棉酚可部分地抑制幼龄大鼠睾丸间质细胞对外源性hCG刺激的反应。主要表现在两方面:间质细胞和成纤维细胞样前体细胞(简称前体细胞,下同)数目减少;以及同质细胞内与类固醇合成有关的酶类(如3β-羟固醇脱氢酶、非特异性酯酶)活性减弱。但光镜下生精上皮及间质细胞的形态在本实验条件下未见明显异常。结果提示,棉酚对睾丸间质细胞的功能有抑制性作用。     

醋酸棉酚在雌性大鼠的抗着床作用及其机理分析

本工作观察了醋酸棉酚对大鼠早期妊娠的影响。于大鼠妊娠的第1—5天,每鼠每天灌服一次醋酸棉酚混悬液(100毫克/公斤体重),对照组灌服等量蒸馏水。于妊娠第10天处死,处死前取血测定血清孕酮,处死后取卵巢测定⊿~5-3β-羟甾脱氢酶活性并记录妊娠动物数及正常着床胎数。结果表明,醋酸棉酚对大鼠具有明显的抗着床作用,抗着床率达85.7%,同时伴有血清孕酮水平下降。若同时皮下注射5毫克孕酮或是同时皮下注射50国际单位的hCG 均能有效地对抗醋酸棉酚的抗着床作用,而同时注射20微克的LH-RH 类似物并不能对抗醋酸棉酚的抗着床作用。在延缓着床的大鼠,醋酸棉酚不能对抗外源性雌二醇的致着床作用。以上结果提示,对抗LH 对黄体功能的支持有可能是醋酸棉酚抗着床作用的机理之一。     

棉酚对大鼠子宫内膜及主要脏器超微结构的影响

本文探讨棉酚作大鼠宫内一次性注射以达到抗早孕目的的同时,观察子宫内膜及内脏细胞的超微结构变化。21只大鼠分实验组和对照组,给药后于不同时间取样作电镜观察。结果显示棉酚对子宫内膜上皮细胞、腺细胞及蜕膜细胞均有不同程度的损伤。但这些损伤引起的超微结构变化,多数是可逆的,可逐步修复的,对内脏细胞基本无毒性反应。根据实验结果结合以往工作,证明棉酚通过一次性宫内给药是一种较安全、有效的抗早孕方法。     

醋酸棉酚对大鼠初级精母细胞DNA含量的影响

本文报导棉酚对大鼠睾丸初级精母细胞DNA 含量的影响和体外抑制DNA合成试验的结果。大鼠服棉酚(30毫克/公斤/日)4及6周后,初级精母细胞核中DNA 的含量较对照组明显减少。其含量分布直方图主峰左移,不再呈典型的正态分布。体外抑制试验表明,当反应液中棉酚浓度为6微克分子时,DNA 合成受抑制率为12%,20微克分子时为20%,60微克分子时抑制率达最高峰值(23%)。根据上述结果,对棉酚干扰生精过程中DNA 代谢及其与抗生育作用的可能关系进行了分析和讨论。     

醋酸棉酚服用者子代外周血淋巴细胞染色体畸变与SCE的观察

本文对服用醋酸棉酚停药恢复生精后所生子代30例及相应的对照组进行了外周血淋巴细胞染色体的观察。实验组的父代每日口服醋酸棉酚20mg,服至起效(一般服药后60～90天以每毫升精液中精子数在400万以下者定为起效指标)。起效后改为维持量,每周40mg。父代平均服药的时间为440天,平均服药总量为3382.34mg。停药后至受孕,时间平均为18个月。结果提示:30例服醋酸棉酚的子代与相应对照组儿童在染色体断裂、裂隙及SCE 均未见明显差异。     

棉酚对大鼠支持细胞蛋白质合成的影响

本实验采用同位素放射自显影方法测定了棉酚对大鼠支持细胞(sertoli tell)蛋白质合成过程中亮氨酸掺入活动的影响。经棉酚在体内外处理的支持细胞与正常支持细胞比较,~(14)C-亮氨酸掺入活动的变化无统计学意义(P>0.05)。提示抗生育剂量棉酚似不影响大鼠支持细胞内的蛋白质合成活动。     

氯化镉对大鼠睾丸波形蛋白表达的影响

目的:探讨氯化镉(CdCl2)对大鼠睾丸曲细精管和间质区波形蛋白表达的影响。方法:20只雄性SD大鼠随机分为对照组与3个实验组,每组5只动物。实验组分别每天腹腔注射CdCl20.2mg/kg(低剂量组),0.4mg/kg(中剂量组)和0.8mg/kg(高剂量组),共14d,对照组注射等量生理盐水。采用免疫组化法检测睾丸波形蛋白的表达和改变。结果:波形蛋白在睾丸曲细精管支持细胞胞浆呈阳性表达。随着Cd2+剂量增加,实验组阳性支持细胞率及波形蛋白表达量与对照组的比较均显著减少(P<0.01),各实验组间比较也有显著性差异(P<0.01)。在中、高剂量组,波形蛋白向管腔伸展的辐射状部分变短,甚至消失。3个实验组的间质区波形蛋白表达均显著低于对照组(P<0.01)。结论:CdCl2染毒可导致大鼠睾丸支持细胞和间质区的波形蛋白表达量显著下降,且损伤呈剂量效应。     

溶脲脲原体感染动物模型的建立─对睾丸的形态学影响

本文观察了溶脲脲原体（Ｕｒｅａｐｌａｓｍａｕｒｅａｌｙｔｉｃｕｍ，Ｕ．Ｕ．）对ＳＤ雄性大鼠睾丸的形态学影响。结果表明，在４０例膀胱内接种Ｕ．Ｕ．（１０５ＣＣＵ／ｍｌ）的大鼠中，有２５例睾丸中分离出Ｕ．Ｕ，动物睾丸重量减轻。光镜下可见１１例动物呈广泛的曲精小管萎缩，生精细胞严重脱落。部分曲精小管仅见支持细胞和少量精原细胞，有的仅剩下界膜。在曲精小管中发现许多多核巨细胞，有多核巨细胞处，生精上皮破坏更为明显，间质渗出、水肿。电镜观察发现，Ｕ．Ｕ．可吸附或侵入生精细胞内，造成生精细胞胞浆空泡化乃至崩解，部分线粒体基质空泡化或嵴消失。其余１４只大鼠睾丸组织有局灶性破坏。     

衰老对小鼠睾丸结构和精子发生的影响

目的:探讨衰老对小鼠睾丸结构和精子发生的影响。方法:昆明小鼠分为老龄组(18月龄,n=15)和青年对照组(6月龄,n=15),取睾丸称重,H.E.染色,观察睾丸组织结构,计数萎缩曲细精管的比例,定量分析睾丸精子发生的状态。结果:老龄小鼠睾丸有部分曲细精管萎缩,多为局灶性,管内生精细胞减少,生精阻滞常发生在初级精母细胞阶段,或有生精细胞脱落至管腔。老龄组萎缩管的比例高于对照组(P<0.01)。定量分析显示老龄组曲细精管横截面内球形精子细胞和延长精子细胞与支持细胞的比例均显著低于对照组(P<0.01),支持细胞数目与对照组相比无显著差异(P>0.05),精原细胞和初级精母细胞与支持细胞的比值也没有显著减少(P>0.05)。结论:老龄小鼠睾丸精子发生衰减,以球形精子细胞和延长精子细胞减少为主。昆明小鼠可作为雄性生殖衰老研究的动物模型。     

雷公藤多甙对大鼠生精细胞及其酶活性的影响

本文连续观察了雄性大鼠服用雷公藤多甙(GTW)30至80d 后睾丸、附睾细胞形态学改变及睾丸 ACP、ALP、3β-HSD、睾丸和附睾生精细胞 LDH-C_4活性的变化;同时注意了药物抗生育作用的可逆性。结果表明:附睾精子先于睾丸生精细胞发生质和量的变化;支持细胞 ACP 活性有增强趋势;睾丸 ALP,LDH-C_4酶活性相对减弱,为生精细胞损伤的结果;用药80d 时,3β-HSD 活性则明显减弱。结果还提示 GTW 引起的不育似有恢复的可能。     

醋酸棉酚对小鼠生殖细胞染色体的影响

本文报道了成熟的ICR 雄性小鼠,每日灌服醋酸棉酚0、1、2.5、5、7.5、10、20、30、40、50毫克/公斤,连续19天后,进行生殖细胞的染色体畸变和SCE 频率的观察。精原细胞的SCE 率检测结果表明,除了20—50毫克/公斤体重/日(临床剂量的50—125倍)高剂量组与对照组比较有显著差异(P<0.005)外,其他用药组与对照组之间不存在明显的差异(P>0.05)。精原细胞和精母细胞染色体异常率的检测表明,各用药组和对照组之间均无明显差异(P>0.05)。结果提示:常用的小剂量棉酚似不具有明显的生殖细胞遗传物质损伤效应。由此推断,停药恢复生殖能力后,对子代可能也不会有明显的影响,但是不可忽视高剂量棉酚引起精原细胞SCE 率的增加。     

Antifertility and ultrastructural effects of optical isomers of gossypol administered intratesticularly in rats

This study reports on the effects of optical isomers of gossypol administered intratesticularly on testicular ultrastructure and fertility in rats. Gossypool isomers were administered to adult male rats by daily intratesticular injections for up to two days at a dose of 1200 ugm/testis. The results of this study have demonstrated that the intratesticular injection of racemic and (-) gossypol at a dose of 1200 ugm for 1 or 2 days mimic the effect of gossypol on rat testis after oral administration of 20 to 30 mg dose for 5-7 weeks; (-)-gossypol was found to be 81% effective in inducing mitochondrial sheath damage in the late spermatids of stage VI, VII and VIII tubules compared to 67% for racemic and 7% for (+)-gossypol, the intratesticular injection of (+/-) and (-)-gossypol (1200 ugm/testis, daily for 1 or 2 days) induces complete infertility three weeks after the last injection.     

Proliferating cell nuclear antigen as a molecular biomarker for spermatogenesis in PTU-induced hypothyroidism of rats

The thyroid hormone has few serious effects on the testes except during the neonatal stage. There is little knowledge concerning the prolonged effect of thyroid hormone deficiency throughout the rat's life span and its effect on spermatogenesis. Proliferating cell nuclear antigen (PCNA) is a nuclear matrix protein, which is essential for multiple cell cycle pathways. Here we used PCNA immunohistochemistry as a marker to differentiate between the testes of control and hypothyroid rats. About 20 rats were equally divided into 2 groups; the first group was the control group, while the second group was the experimental group in which rats were fed 0.05% 6-n-propyl thiouracil (PTU) in drinking water for 6 weeks. Immunohistochemistry, using an antibody against PCNA, showed at least 3 differences in the pattern of PCNA immunoreactivity (PCNA-ir). First, PCNA-ir was not detected in Sertoli and Leydig cells in the testes of control rats and detected in some of the hypothyroid rats. Second, in the control group more than 96% of spermatogonia were PCNA-positive cells; however, hypothyroidism caused the reduction to approximately 25% PCNA staining in spermatogonia. The third difference was in the abnormal distribution of spermatogonia seen in the hypothyroid rat testis, not in the control one. These results suggest that prepubertal hypothyroidism affects the proliferation of spermatogenic cells leading to impaired spermatogenesis and that PCNA index is a useful marker for assessing germ cell kinetics and spermatogenesis in prepubertal hypothyroidism.     

Antigens of the basement membranes of the seminiferous tubules induce autoimmunity in Wistar rats

A preparation enriched in basement membranes from seminiferous tubules was isolated from rat testes (STBM) and injected with complete Freund's adjuvant into Wistar rats. In 60% of the animals a mild multifocal orchitis was observed. In damaged areas, perivascular and peritubular mononuclear cell infiltrates and different degrees of cell sloughing of some seminiferous tubules were observed. Electron microscopy revealed focal thickenings and delamination of the basement membrane of the seminiferous tubules as well as vacuolization of Sertoli cell cytoplasm. Using immunofluorescence discontinuous linear deposits of IgG were detected along the seminiferous tubular wall. Moreover, the same pattern of immunofluorescence was observed when the IgG eluted from the testes of the immunized rats was layered on sections of normal rat testis. Circulating antibodies to STBM were detected using passive haemagglutination in approximately 45% of the immunized rats, with titers ranging from 1:20 to 1:80. Leukocyte migration was inhibited when the spleen cells of the immunized rats were incubated with antigens from the basement membrane of seminiferous tubules, whilst a negative reaction was obtained when the soluble fraction of testis homogenate was used.     

Testicular involution in elderly men: comparison of histologic quantitative studies with hormone patterns

An endocrinologic and quantitative histologic study was carried out in 64 elderly men who underwent orchidectomy owing to prostatic carcinoma. The men were classified into age groups (decade of life), and each group was subdivided into group A (testes with complete spermatogenesis in most tubules) and group B (testes showing maturation arrest of spermatogenesis in most tubules). Up to 80 years of age, men of group A showed hormone levels and testicular parameters similar to those of young control men. From 50 to 60 years of age, men of group B showed a significant decrease in testicular volume, tubular volume, tubular length, number of germ cells, Sertoli cells and Leydig cells per testis, and plasma testosterone levels, whereas the tunica propria thickness and plasma levels of both follicle-stimulating hormone and luteinizing hormone were increased.