不同阶段生精小管组织学结构研究

目的探讨不同阶段人体睾丸生精小管面积、生精小管管腔面积变化和生精上皮在不同阶段的组织学特点,及其与生殖的关系。方法:应用常规组织制片技术和图像分析技术。结果①生精小管平均面积变化,从胚胎睾丸形成到青春期前,随睾丸逐渐发育增大,睾丸间质增多,但生精小管面积无明显增大;自青春期生精小管面积迅速增大,25岁左右达到峰值,45岁左右生精小管平均面积缓慢减少,55岁以后显著减少。②生精小管管腔面积变化,从胚胎睾丸形成到青春期前,生精小管几乎无管腔;青春期管腔开始出现并迅速增大,20岁左右达到峰值;于45岁左右管腔面积缓慢减少,55岁以后显著减少。③生精小管的组织学结构变化,从胚胎睾丸形成到青春期前,生精小管上皮由精原细胞和支持细胞组成,但随睾丸发育增大,睾丸间质增多,生精小管上皮和基膜间渐出现明显的间隙;青春期开始,生精小管上皮发育,生精细胞层数增加,管壁各级生精细胞典型,腔面可见精子;55岁后睾丸纤维化明显,生精小管皱缩,随年龄增长,生精上皮细胞数量渐减少,排列紊乱,基膜增厚。结论①生精细胞增殖旺盛是生精小管平均面积迅速增大的原因之一;生精细胞增殖旺盛的阶段是20～30岁,最佳时期是25岁左右。②生精小管管腔的出现与生精细胞的凋亡、基膜扩大的速率远远大于生精细胞的增殖水平有关,而生精小管管腔的出现有利于精子的生成与运输。③衰老睾丸生殖功能的下降与其组织结构的纤维化及生精小管基膜厚度增加等因素有关。     

环磷酰胺对大鼠睾丸和附睾的影响

目的探讨抗肿瘤药物所致大鼠睾丸和附睾损害,为寻找男性不育症中药治疗的研究提供理论依据。方法选用16只15周龄SD大鼠随机分为对照组和实验组,每组8只;实验组腹腔注射环磷酰胺20mg·kg-1·d-1,连续5d,用药2个月后,应用HE染色法研究大鼠睾丸、附睾远期组织学变化,用原位缺口末端标记法(TUNEL方法)检测生精细胞凋亡。结果实验组大鼠体重、睾丸和附睾重量均显著减轻(P<0.01),睾丸生精小管直径缩小、间距增宽、生精上皮变薄、生精细胞层次和数量减少、生精小管腔多未见精子形成,实验组睾丸生精小管直径、面积、生精上皮细胞数、均显著低于对照组(P<0.01);实验组生精细胞凋亡增多,与对照组比较,生精细胞显著凋亡(P<0.01);附睾管管腔内腔内精子稀少,含有大量脱落细胞,管壁变薄。结论环磷酰胺对大鼠睾丸、附睾远期损害明显,促进生精细胞凋亡。     

模拟高原缺氧环境对大鼠生殖细胞凋亡的影响

目的观察模拟高原缺氧环境条件对雄性SD大鼠生殖细胞形态及凋亡的影响。方法随机将20只SD雄性大鼠分为两组,对照组置650m海拔高度饲养,低氧组置模拟6000m海拔高度低压氧舱饲养。饲养14天后脱颈处死大鼠,采集睾丸组织标本,HE染色观察睾丸组织形态学变化。采用TUNEL法检测高原缺氧大鼠生殖细胞凋亡的影响。结果与对照组比较,可见高原缺氧组大鼠睾丸组织生精小管间隙增宽,部分生精小管中初级精母细胞、次级精母细胞、精细胞减少,间质血管充血、扩张,部分生精小管内精细胞数量明显减少,初级、次级精母细胞变性、脱落、坏死。高原缺氧组睾丸内发生生殖细胞凋亡的生精小管数量[每100个生精小管中含有1、2、3及3个以上凋亡细胞的小管数分别为(6.80±1.40)、(3,20±0.38)、(2.80±0.60)个]均显著的高于常氧对照组[每100个生精小管中含有1、2、3及3个以上凋亡细胞的小管数分别为(3.70±0.69)、(1.40±0.56)、(0.60±0.48)个,P0.05]。结论高原低氧环境可能是通过诱发生殖细胞凋亡途径,导致大鼠睾丸生殖机能损伤。     

饮酒对小鼠睾丸生精小管一氧化氮合酶、增殖细胞核抗原表达及细胞凋亡影响的研究

目的　探讨酒精对小鼠睾丸的组织结构、内皮型一氧化氮合酶 (eNOS)、增殖细胞核抗原 (PCNA)及细胞凋亡的影响。　方法　用 5 %、10 %及 15 % 3种不同浓度的酒精作用于 2 2d龄小鼠 ,取睾丸做石蜡切片、HE染色 ;用免疫组织化学方法检测睾丸eNOS、细胞增殖的变化 ;TUNEL法检测细胞凋亡的变化 ,并进行统计学分析。　结果　随着酒精浓度的增大 ,睾丸组织结构发生明显改变 ,生精小管的直径逐渐减小 ,eNOS阳性细胞面积密度逐渐增大 ,单位面积内PCNA阳性细胞和凋亡细胞数目增加 ,高浓度酒精组与其他组差异极显著 (P <0 0 1)。　结论　酒精可使生精小管的直径变小 ,eNOS及PCNA表达增强 ,凋亡细胞增加并随酒精浓度的增大而变化加重。这可能是过量饮酒导致生精细胞减少 ,生殖能力降低的重要因素。     

葛根素减轻乙醇导致大鼠生精细胞的凋亡

目的 观察乙醇导致大鼠生精细胞的凋亡及葛根素的干预。方法 将大鼠30 只,随机均分为对照组、乙醇组及葛根素干预组。于实验第40天免疫组织化学法（SABC）检测左侧睾丸各组Bcl-2、Bax 蛋白在生精细胞的表达; RT-PCR检测右侧睾丸各组Bcl-2及Bax mRNA的表达;TUNEL 法检测生精细胞的凋亡。结果 醇组平均每个生精小管断面中Bcl-2 蛋白阳性细胞数和A值低于对照组(P<0.01),而平均每个生精小管断面中Bax 蛋白的阳性细胞数和A值高于对照组(P<0.01);乙醇组Bax mRNA表达较葛根素干预组及对照组强（P<0.05）,而Bcl-2 mRNA表达较葛根素干预组及对照组弱（P<0.05）;乙醇组每个生精小管横切面中的凋亡细胞数目高于对照组(P< 0.01),葛根素干预组显著缓解上述变化。结论 根素对乙醇导致的大鼠生精细胞凋亡有干预作用。     

睾丸动脉血流障碍对小鼠睾丸生精上皮和血-睾屏障的影响

目的:探讨睾丸动脉血流障碍对小鼠睾丸生精上皮和血-睾屏障的影响.方法:成熟雄性BALB/c小鼠,随机分为对照组和6个实验组,对照组仅暴露睾丸动脉而不进行结扎,实验组显微镜下行单侧睾丸动脉结扎, H-E染色观察睾丸组织的病理改变,电镜观察血-睾屏障的变化.结果:对照组睾丸生精小管无明显病理改变;1h组睾丸生精小管上皮细胞排列不均匀,细胞界限清楚,线粒体轻微肿胀,核周隙轻微增宽,睾丸生精小管的基膜完整;2h和3h组生精小管生精细胞层数减少,线粒体轻微空泡改变,线粒体嵴扩张,支持细胞内质网扩张明显,有局部液化灶,生精小管基膜轻微波纹状;睾丸间质可见炎性细胞浸润,多为分叶的中性粒细胞;4h和5h组睾丸间质血管中充血明显,血管内壁有大量的炎性细胞贴壁现象,血管壁增厚呈玻璃样变,精原细胞有核碎裂现象,细胞界限不清,基膜间隙增宽,基膜皱折,精子细胞顶体位置未见顶体,精子细胞局部有空泡,界限不清,细胞膜不完整;6h组睾丸生精小管内主要为精原细胞和初级精母细胞,精原细胞与基膜脱离,基膜有"分层"现象,支持细胞、精母细胞肿胀明显,细胞膜不完整,界限不清,生精小管内可见多核巨细胞.结论:单侧睾丸动脉结扎/再通可引起睾丸生精细胞的缺血性损伤和坏死,生精上皮细胞和血-睾屏障的病理变化随缺血时间的延长而加重.     

慢性镉中毒小鼠生精上皮的超微结构观察和立体定量分析

本用透射电镜观察并结合立体定量法研究了慢性镉中毒时小鼠睾丸生精上皮的超微结构。结果为：（1）支持细胞明显受损，细胞核增大，细胞器出现变性变化，有的支持细胞呈现坏死现象，（2）各级生精细胞有不同程度损伤，生精小管管腔内常见未成熟精子和畸形精子。（3）支持细胞的细胞连接被破坏，血－生精小管屏障部分瓦解，本还对慢性镉中毒时影响精子发生的机理进行了初步探讨，认为支持及其细胞连接的结构和功能异常可能是生精障碍的主要原因。     

DahlS高血压大鼠生精细胞凋亡及Bcl-2蛋白的表达

目的　为阐明高血压导致生精障碍的确切机制。方法　应用原位末端标记法 (TUNEL)、免疫组织化学、流式细胞检测及电镜等方法 ,对DahlS高血压大鼠生精细胞凋亡的定性、定位、定量及其相关蛋白Bcl 2的表达进行了研究。结果　实验组随高血压病程延长 ,血压升高 ,生精细胞凋亡率增高 ,15、17周龄明显高于对照组 (P <0 0 5 )。Bcl 2蛋白表达降低 ,15、17周龄与对照组相比差异显著 (P <0 0 5 )。随糖尿病病变加重 ,Bcl 2表达减少 ,而且 ,糖尿病组比对照组Bcl 2表达率低。随糖尿病病程进展和年龄增加 ,表达均呈增加趋势 ,而且 ,糖尿病组比对照组Bax表达率高。结论　高血压可导致凋亡抑制基因Bcl 2表达降低 ,从而使生精细胞凋亡增加 ,这可能是高血压导致生精障碍的机制之一     

老化大鼠睾丸的形态学研究

应用光镜、电镜及放免技术,对青年、初老及老年SD大鼠的睾丸进行了观察。随增龄,睾丸重量减轻,生精小管直径缩小。初老时精细胞疏松、脱落呈团块状阻塞于管腔内,多核巨细胞出现,老年时生精细胞缺如仅剩支持细胞或见散在分布,生精小管萎缩并呈空泡变、玻璃样变及钙化。衰老时支持细胞内出现很多脂褐素及髓样小体,线粒体及内质网扩张。间质细胞内线粒体及滑面内质网也呈老化改变。表明:大鼠随着年龄的增长睾丸出现明显的衰老变化。     

细辛对D-半乳糖所致衰老小鼠睾丸的形态学研究

本文研究了细辛对D－半乳糖所致衰老小鼠的抗衰老作用，应用光镜电镜技术测定小鼠睾丸的重量、生精小管直径、生精上皮细胞数、随龄变化的间质细胞数，同时观察了细辛对上述指标的影响。结果 1．随增龄、睾丸重量减轻，生精小管直径缩小。衰老时租精细胞缺如，仅有支持细胞或散在分布，间质细胞随龄递减。2．细辛可以使小鼠的曲细精管增粗，生精过程活跃，生精细胞增多，间质细胞增多。结论 细辛有一定抗衰老作用。     

Trypanosoma cruziand myoid cells from seminiferous tubules: interaction and relation with fibrous components of extracellular matrix in experimental Chagas' disease

The main transmission route of Trypanosoma cruzi is by triatomine bugs. However, T. cruzi is also transmitted through blood transfusion, organ transplantation, ingestion of contaminated food or fluids, or is congenital. Sexual transmission, although suggested since the discovery of Chagas' disease, has remained unproven. Sexual transmission would require T. cruzi to be located at the testes and ovaries. Here we investigated whether T. cruzi is present in the gonads of mice infected with 10⁴ T. cruzi trypomastigotes from the CL strain. Fourteen days after experimental infection, histopathological examination showed alterations in the extracellular matrix of the lamina propria of the seminiferous tubules. Furthermore, amastigotes were present in seminiferous tubules, within myoid cells, and in the adjacencies of the basal compartment. These results indicate that T. cruzi is able to reach seminiferous tubule lumen, thus suggesting that Chagas' disease could potentially be transmitted through sexual intercourse. Complementary studies are required to demonstrate that Chagas' disease can be transmitted by coitus.     

Ultrastructure of germ cells and sertoli cells in the postnatal rabbit testis

Spermatogenesis begins in the rabbit at seven to eight weeks of age. During the preceding postnatal period, seminiferous tubules contain round to oval prespermatogonia arranged in pairs at the tubular periphery and elongated Sertoli cells lying perpendicular to the basal lamina. Prespermatogonia are distinguished from fetal gonocytes by their abundant cytoplasm and the presence of intercellular bridges. Preceding the onset of spermatogenesis, prespermatogonia undergo diminution in cell size and cytoplasmic condensation. At the same time there is marked nucleolar enlargement, with elaborate proliferation of the nucleolonema. By the end of the seventh week, characteristic type A and B spermatogonia are present. Spermatocytes are seen at the end of the eighth week. Spermatids appear in the twelfth week, coincident with tubular lumen formation. The duration of the initial spermatogenic cycle is similar to that in the adult. Sertoli cells are closely associated with adjacent germinal elements throughout the postnatal period. The Sertoli cell cytoplasm includes elongated mitochondria, abundant smooth endoplasmic reticulum and prominent Golgi complexes. Many of the cells contain phagocytized material, presumably derived from degenerating germ cells. The observations suggest possible mechanisms by which Sertoli cells support early germ cell maturation.     

Morphological and histochemical characteristics of the lamina propria in scrotal and abdominal testes from postpubertal boars: correlation with the appearance of the seminiferous epithelium

This study was undertaken to investigate the morphological characteristics and lectin affinity of the testicular lamina propria in healthy boars and in unilateral and bilateral abdominal cryptorchid boars. The lamina propria of scrotal testes from healthy boars and unilateral cryptorchid boars was constituted by an innermost noncellular layer, the basal lamina, and by 2 layers of peritubular cells, each separated by a fibrous layer. The noncellular layers contained collagen fibres and glycoconjugates with abundant N‐acetylgalactosamine, galactose, fucose, N‐acetylglucosamine and neuraminic acid residues. The inner peritubular cell layer was composed of myoid cells, the outer layer of fibroblasts. In the abdominal testes of unilateral and bilateral cryptorchid boars, the lamina propria of nondegenerating and degenerating seminiferous tubules appeared thickened due to an increased content of collagen fibres and glycoconjugates. Glycoconjugates showed decreased amounts of fucose, neuraminic acid and galactose, and increased amounts of N‐acetylglucosamine residues. The basal lamina formed infoldings toward the seminiferous epithelium and contained small cells. Both inner and outer peritubular cells were fibroblasts of immature appearance. In degenerated seminiferous tubules of bilateral cryptorchid boars, the lamina propria was composed of a thickened and collagenised basal lamina, without peritubular cells and with a low content of glycoconjugates. In scrotal testes, therefore, the lamina propria was implicated in tubular contractility and in mediating the communication and the substrate diffusion between seminiferous tubules and interstitial tissue. Cryptorchidism induced morphological and histochemical alterations in the lamina propria of abdominal testes, which may be linked to evidence from other studies of lack of tubular contractility and defective cell–cell communication and substrate diffusion. The severity of these anomalies correlated with the severity of Sertoli cell alterations.     

生后发育过程中睾丸神经生长因子基因的表达

为了通过观察小鼠生后发育过程中睾丸组织神经生长因子 ( NGF)基因表达的变化藉以探讨 NGF在睾丸发育、精子发生过程中的生物学作用 ,本研究采用 RT-PCR方法 ,半定量分析生后 1d、1周、2周、4周及 8周小鼠睾丸组织中 NGF m RNA的表达变化 ;以地高辛标记的βNGF c DNA为探针 ,采用原位杂交法观察 NGF m RNA在幼年及成年小鼠睾丸组织中的分布。 RT-PCR结果显示 ,小鼠生后 1d、1周、2周、4周、8周睾丸组织中均有 NGF m RNA的表达 ,以生后 1周时的表达量为最高。原位杂交结果表明 ,幼年鼠 NGF m RNA杂交信号位于睾丸的间质之中 ,而成年鼠 NGF m RNA杂交信号则主要位于睾丸生精小管管壁和管腔中。结果提示 :NGF m RNA在小鼠出生时即有表达 ,其表达量及在睾丸组织中的分布 ,随小鼠的发育而有变化     

Macro-orchidism: light and electron microscopic study of four cases.

A hormonal and quantitative light microscopy study of one man with macro-orchidism associated with mental retardation and fragile X chromosome (case no. 1) and three men with idiopathic macro-orchidism (cases no. 2 to 4) is reported. Hormonal study revealed slightly increased follicle-stimulating hormone serum levels in cases no. 1 to 3. The testes from cases no. 1 (orchidoepididymoectomy specimen) and 2 (testicular biopsy) presented interstitial edema and three different tubular patterns that were arranged in a mosaic-like manner. Type I tubules had an increased diameter (less than 220 microns), dilated lumen, and thin seminiferous epithelium usually consisting of Sertoli cells, spermatogonia, primary spermatocytes, and sometimes a few spermatids. Type II tubules had a normal diameter (180 to 220 microns) and germ cell development varied between complete spermatogenesis and Sertoli-cell-only tubules. Type III tubules had decreased diameter (less than 180 microns), atrophic seminiferous epithelium, and thickened tunica propria. The appearance of the nuclei of the Sertoli cells in the three types of tubules could be either mature or immature. Some of the mature Sertoli cells presented a granular cytoplasm. A few of these granular cells grouped together, forming nests that protruded into the tubular lumen. The testicular biopsies from cases no. 3 and 4 only presented type II tubules that contained both mature and immature Sertoli cells. Quantitative study revealed that the large testicular size was principally due to an increased tubular length in all four cases. Although the seminiferous tubule lesions and interstitial edema suggest an obstructive process, the testicular excretory ducts (studied in case no. 1) appeared normal or only slightly dilated. It is possible that the seminiferous tubule lesions (dilated lumen and germ cell depletion) might be secondary to the Sertoli cell lesions (granular cytoplasm and nuclear immature-like pattern.     

实验性隐睾术后大鼠睾丸生精上皮酶的动态变化

目的：研究热作用对睾丸生精过程影响中AKP、ACP和SDH活性的变化,探讨其生物学意义,为男性抗生育提供形态学资料。方法：人工隐睾术后1－70天取出大鼠睾丸,显示AKP、ACP和SDH活性,光镜观察。结果：术后曲细精管界膜AKP活性减弱,直到晚期仍较弱;支持细胞术后1－10天酶增强,随后减弱,50天始为微弱反应;各级生精细胞反应不一。术后界膜ACP阴性;支持细胞酶增强,晚期仍较强;生精细胞酶反应不一。术后界膜SDH为阳性。支持细胞早期反应较强,3天始减弱,到晚期仍较弱;生精细胞酶减弱,15天始为阴性。结论：隐睾术后热作用影响生精过程,使AKP、ACP和SDH有明显变化,因各级生精细胞对热作用的反应性与耐受性不同,使以上三种酶反应强弱不一。     

小鼠肾脏发育中的细胞凋亡

目的：研究小鼠肾脏发育过程中的细胞凋亡规律及形态学特点,方法：应用光镜、电镜技术和TUNEL法分别对不同胚龄、生后日龄小鼠肾脏细胞凋亡进行了观察。结果：皮质凋亡细胞多出现在生肾区S小体之间和是肾小体内,凋亡高峰期在胚龄14－18d之间,髓质凋亡细胞出现在肾小鼠管上皮内,凋亡高峰期在生后7d 左右。超微结构观察可见皮质和髓质凋亡细胞主要表现为核固缩,染色质凝集,细胞皱缩。皮质和髓质凋亡细胞结局为,被邻近细胞吞噬,或脱落到肾小管腔内,结论：小鼠肾脏发育过程中确有细胞凋亡;皮质中细胞凋亡与生肾区的出现和肾小体发育完善有关,髓质中细胞凋亡与髓质中肾小管和集合小0管的有发育完善有关。     

Immunohistochemical study of flotillin-1 in the rat testis during postnatal development

The level and cellular localization of fotillin-1, a lipid raft protein, was examined in the testis of rats during postnatal development and spermatogenesis in order to determine if flotillin-1 is involved in testicular development. The testes of rats were sampled on postnatal days 7, 14, 21, 40, and 60, and analyzed by Western blot and immunohistochemistry. Western blot analysis detected flotillin-1 in the testes at days 7 and 14 after birth but the level decreased significantly at postnatal days 21, 40 and 60. At postnatal days 7, 14, 21, and 40, flotillin-1 immunolocalization was observed mainly in the Sertoli cells. However, there was little flotillin-1 immunolabeling in the spermatogenic cells from the seminiferous tubule of the testes. In the seminiferous tubule of the testes at postnatal day 60, flotillin-1 immunoreactivity in the Sertoli cells varied according to the stages of the spermatogenic cycle; intense immunoreactivity being observed in stages IX–III and less in stages IV–VIII. These results suggest that flotillin-1 participates in the developmental process of Sertoli cells and is involved in the regulation of spermatogenesis.