细辛、杜仲及其合剂对亚急性衰老小鼠睾丸及血清睾酮影响的实验研究

目的研究细辛、杜仲及其合剂对亚急性衰老小鼠睾丸、精子及血清睾酮的影响。方法选昆明系雄性小鼠,用D-半乳糖制备衰老模型。应用光镜、电镜技术检测细辛、杜仲及其合剂治疗前后衰老小鼠睾丸及精子的形态学改变;放射免疫技术检测血清睾酮的变化。结果衰老小鼠睾丸重量减轻,生精细胞缺如,精子密度及活动率下降,血清睾酮含量明显降低(模型组与青年组比较:P<0.01);用药后,睾丸重量增加,生精小管增粗,生精细胞增多,生精过程活跃,精子密度及活动率明显提高,血清睾酮含量增加(P<0.05)。合剂组的作用优于杜仲组和细辛组(P<0.05)。结论细辛,杜仲及其合剂可改善衰老小鼠的生精功能,明显抑制衰老小鼠血清睾酮含量的下降,具有一定延缓衰老的作用。     

延衰合剂对D-半乳糖所致亚急性衰老小鼠睾丸及血清睾酮的影响

目的 观察延衰合剂(YSM)对D-半乳糖所致亚急性衰老小鼠睾丸及血清睾酮的影响.方法 选昆明系雄性小鼠,用D-半乳糖建立衰老模型.应用光镜、电镜技术观察延衰合剂治疗前后衰老小鼠睾丸的形态学改变;酶联免疫分析法检测血清睾酮的变化.结果 衰老小鼠生精细胞减少,生精小管变细,血清睾酮含量明显降低;用药后,生精小管增粗,生精细胞增多,间质细胞增多,血清睾酮含量增加(P<0.05).延衰合剂高剂量组作用优于补肾益寿胶囊组及延衰合剂低中剂量组(P<0.05).结论 延衰合剂可以改善衰老小鼠睾丸的形态结构,抑制衰老小鼠血清睾酮含量的下降,具有一定的延缓衰老的作用.     

细辛、杜仲及其合剂对D—半乳糖所致衰老小鼠NO、NOS和CAT的影响

目的 研究细辛、杜仲及其合剂对D－半乳糖所致衰老小鼠的抗衰老作用。方法 观察小鼠脑组织一氧化氮合酶（NOS）、血浆一氧化氮（NO）、全血过氧化氢酶（CAT）的随龄变化,同时观察细辛、杜仲及基合剂对上述指标的影响。结果 老龄小鼠血浆NO含量,脑组织NOS、全血CAT活性在中年期以前随龄上升,中年期以后随龄下降;细辛、杜仲及基合剂能够增加NO含量,提高NOS、CAT活性。结论 细辛、杜仲及其合剂具有一定的抗衰老作用。     

睾丸的老化

机体的老化是以性功能变化为特征的。随着年龄的增长,睾丸的重量和体积下降,曲精小管直径减小,基膜增厚,生精细胞减少乃至丧失。同时支持细胞和间质细胞的数量减少,结构和功能出现明显的老化特征。间质内结缔组织也随增龄发生纤维化及进行性玻璃样变。睾丸内琥珀酸脱氢酶及3-β羟基类固醇脱氢酶等酶活性降低。这些改变最终导致睾丸功能减退及机体衰老。     

蒙药索吉德-11对催老小鼠睾丸功能的影响

目的 研究蒙药索吉德-11对催老小鼠睾丸功能的影响.方法 1月龄的ICR小鼠分为4组,正常对照组和衰老模型组用生理盐水灌胃,给药组用索吉德-11分别以每日0.2、0.1 g/kg的剂量灌胃,连续灌胃6W后检测小鼠血清睾酮含量、睾丸组织丙二醛(MDA)含量、超氧化歧化酶(SOD)的活性及睾丸生精细胞核抗原(PCNA)表达的变化情况.结果 索吉德-11可提高衰老小鼠血清睾酮含量(P<0.05)、睾丸组织SOD的活性及睾丸生精细胞PCNA表达水平(P<0.05),并降低睾丸组织MDA含量.结论 蒙药索吉德-11可明显改善衰老小鼠睾丸的功能,具有一定延缓衰老的作用.     

大鼠睾丸微血管变化的定量研究

目的 探讨大鼠睾丸微血管的增龄变化及其对睾丸功能的影响。方法 采用石蜡切片及HE染色法，对睾丸微血管进行定量测定。结果 随着增龄变化，微血管管壁的平均厚度呈增大趋势，从初生到2月龄每个生精小管周围的微血管数目不断增加，到6月龄略减少，12月龄至24月龄微血管数量呈减少趋势，单位面积内微血管数目与生精小管数目的比值从初生到6月龄增至最大值，到24月龄比值呈减小趋势。结论 微血管管壁厚度与微血管数目变化是睾丸衰老的重要原因。     

天年饮对亚急性衰老大鼠睾丸生精细胞增殖与凋亡的影响

目的　观察天年饮对亚急性衰老大鼠血清超氧化物歧化酶 (SOD)活性、睾酮含量及睾丸生精细胞增殖与凋亡的影响。方法　 D-半乳糖连续腹腔注射制作亚急性衰老大鼠模型 ,检测各组大鼠天年饮灌胃前后血清 SOD的活性、睾酮的含量以及睾丸生精细胞增殖和凋亡的情况。结果　 D-半乳糖致亚急性衰老大鼠血清 SOD活性、睾酮含量及睾丸生精细胞 PCNA阳性表达率均明显下降 ,与正常组比较差异显著 (P<0 .0 1 ,P<0 .0 5) ;经天年饮灌胃后 ,三者均有明显提高。 D-半乳糖致亚急性衰老大鼠睾丸生精小管凋亡百分率及凋亡阳性细胞率均增高 ,与正常组比较差异显著 (P<0 .0 5) ;经天年饮灌胃后 ,二者均降低。结论　天年饮可显著提高亚急性衰老大鼠血清 SOD活性、睾酮含量及睾丸生精细胞 PCNA的表达水平 ,降低睾丸生精细胞的凋亡指数 ,因此 ,天年饮具有延缓性腺衰老的作用。     

蒺藜皂苷对环磷酰胺致小鼠生精细胞凋亡的保护作用

目的研究蒺藜皂苷对环磷酰胺(cyclophosphamide,CP)所致小鼠生精细胞凋亡的保护作用,探讨蒺藜皂苷对生殖功能障碍小鼠的保护机制。方法将8周龄的雄性昆明种小鼠随机分为3组,对照组采取腹腔注射生理盐水,模型组和治疗组采取连续5d腹腔注射CP 80mg/(kg·d),腹腔注射同时将对照组和模型组小鼠连续灌胃生理盐水,治疗组小鼠连续灌胃蒺藜皂苷200mg/(kg·d)35d。比较各组小鼠体质比及睾丸重量;检测附睾精子密度、活精子百分率、精子活力和精子畸形率;HE染色观察睾丸生精上皮的发育;TUNEL法检测生精细胞的凋亡;Western blot检测睾丸组织中凋亡相关蛋白Caspase-8表达水平的变化。结果与模型组比较,蒺藜皂苷显著提高了生精功能损伤小鼠的精子密度、活精子百分比、精子活力,降低了精子畸形率,促进了生精小管上皮精原细胞及精子细胞的发育,上调了睾丸组织中Caspase-8的表达水平,使生精细胞的凋亡减少。结论蒺藜皂苷能显著修复CP所致的小鼠睾丸损伤,其修复机制可能通过上调Caspase-8的表达,拮抗环磷酰胺引起的生精功能损伤,减少生精细胞的凋亡。     

人参总皂甙抗衰老作用的实验研究

应用衰老模型小鼠对人参总皂甙的抗衰老作用进行了实验研究，结果表明：人参总皂甙能增加衰老模型小鼠的胸腺、肾上腺、睾丸及卵巢等脏器系数并减少衰老模型小鼠的肝脏和脾脏系数，人参总皂甙还能明显提高衰老模型小鼠的海马和血清中的超氧化物歧化酶含量，降低其过氧化脂质的代谢产物丙二醛含量，以上结果提示人参总皂甙能减少自由基对细胞的损伤，衰老模型小鼠的免疫及内分泌等多种功能，具有一定的抗衰老作用。     

何首乌饮对衰老大鼠生精细胞凋亡的影响

目的探讨何首乌饮对衰老大鼠睾丸生精细胞凋亡的影响。方法选用18月龄Wistar大鼠为自然衰老动物模型,以12月龄大鼠作为青年对照,采用原位末端标记法和流氏细胞术分别检测何首乌饮灌胃用药60 d和30 d的18月龄大鼠睾丸组织细胞凋亡率变化和DNA含量的变化。结果与青年对照组相比,自然衰老组睾丸细胞凋亡率明显增高,单倍体和4倍体细胞明显减少并以二倍体为主,G_1/G0期细胞明显增多(P<0.05),细胞增殖指数明显降低(P<0.05);而何首乌饮用药后与自然衰老组相比,细胞凋亡率明显降低(P<0.05);4倍体和单倍体比例明显增加(P<0.05),与青年对照组无显著性差异(P>0.05);细胞增殖指数明显升高(P<0.05),G_1/G_0期细胞明显减少(P<0.05)。结论何首乌饮可以抑制生精细胞凋亡,促进细胞增殖,达到延缓睾丸组织衰老的效果,但其具体作用机制有待研究。     

康欣口服液对老年小鼠睾丸超微结构的影响

目的 研究康欣口服液对老年Balb/c小鼠睾丸超微结构的影响。方法 Balb/c小鼠随机分为青年组、老年组和老年康欣口服液组,分别给予生理盐水和康欣口服液4个月,采用透射电镜观察小鼠睾丸超微结构。结果 青年组睾丸超微结构未见明显异常,老年组睾丸生精细胞明显减少,分裂异常,呈坏死脱落。支持细胞内线粒体肿胀,间质细胞内次级溶酶体明显增多。与老年组相比,老年康欣口服液组睾丸超微结构损伤有明显改善。结论 康欣口服液可改善老年小鼠睾丸超微结构损伤。     

Lifelong running reduces oxidative stress and degenerative changes in the testes of mice

Regular exercise can counteract the adverse effects of aging on the musculoskeletal and cardiovascular systems. In males, the normal aging process is associated with reductions in testosterone production and impaired spermatogenesis, but the underlying mechanisms and their potential modification by exercise are unknown. Here, we report that lifelong regular exercise (running) protects the testes against the adverse effects of advancing age, and that this effect of running is associated with decreased amounts of oxidative damage to proteins, lipids, and DNA in spermatogenic and Leydig cells. Six-month-old male mice were divided into a sedentary group and a group that ran an average of 1.75 km/day, until the mice reached the age of 20 months. Seminiferous tubules of runners exhibited a full complement of cells at different stages of the spermatogenic process and a clear central lumen with large numbers of spermatozoa, in contrast to sedentary mice that exhibited disorganized spermatogenic cells and lacked spermatocytes in a central lumen. Levels of protein carbonyls, nitrotyrosine, lipid peroxidation products, and oxidatively modified DNA were significantly greater in spermatogenic and Leydig cells of sedentary mice compared with runners. These findings suggest that lifelong regular exercise suppresses aging of testes by a mechanism that involves reduced oxidative damage to spermatogenic and Leydig cells.     

Follicle-stimulating hormone induction of Leydig cell maturation

The effects of FSH on the testes of immature hypophysectomized rats were investigated by comparing functional changes in Leydig cells with changes in their number and morphological appearance. Rats were treated twice daily for 7 days with 0.5 ml saline vehicle, 10 micrograms rat FSH, or 20 ng ovine LH (an equivalent amount of LH known to contaminate the FSH preparation). FSH treatment caused a significant increase in testis weight and stimulated more advanced spermatogenic activity compared to saline or LH treatment. Morphometric analysis of glutaraldehyde perfusion-fixed testes showed no significant increase in Leydig cell number after LH treatment [saline, 4.63 +/- 0.14 million cells; LH, 6.38 +/- 0.55 million mean +/- SE)], but a significant (P less than 0.001) increase after FSH treatment (11.55 +/- 0.79 million). FSH, but not LH or saline, treatment resulted in Leydig cell hypertrophy and ultrastructural features identical to those of adult Leydig cells, these changes being reflected by enhanced hCG- and LHRH agonist-stimulated testosterone production in vitro by whole testes or dispersed interstitial cells. FSH and LH treatment caused minor but significant decreases in LH receptor numbers on dispersed interstitial cells compared to saline treatment. LHRH receptor numbers on interstitial cells were significantly increased only by FSH treatment. It is suggested that since FSH acts only on the seminiferous epithelium, then the hypertrophy, hyperplasia, and functional enhancement of Leydig cells after FSH treatment may be mediated by the secretion of one or more factors from the seminiferous tubules, providing additional evidence to support the view that gonadotropic regulation of testicular function is modulated by local interactions between the seminiferous tubules and the interstitial tissue.     

Testicular plasminogen activators during postnatal development in the rat

In the seminiferous epithelium, Sertoli cells secrete plasminogen activator (PA) under regulation of follicle stimulating hormone, cyclic AMP and neighbouring spermatogenic cells. Recent observations suggest that preleptotene spermatocytes upon their release from the basement membrane of the seminiferous tubule are important regulators of PA secretion. To study further the role of PA's in the seminiferous tubules, we have analyzed the endogenous levels and secretion rates of PA at various ages during postnatal development, and performed biochemical analyses of the types of PA in the testis and spent media from seminiferous tubular cultures. Cyclic secretion of PA started at the age of 28 days, and from 40 days onwards, the high secretion rates were localized in stages VII and VIII of the cycle of the seminiferous epithelium. The secreted PA is most obviously of the urokinase type; both urokinase-type and tissue-type PA-like activities were found in seminiferous tubular homogenates. The increase in testicular PA levels concomitant to the onset of meiosis in the epithelium was due to the urokinase-type PA-like activity.     

The differentiation of Leydig cells, steroidogenesis, and the spermatogenetic wave in the testis of Necturus maculosus

The study of seminiferous tubule--Leydig cell interactions in relation to specific germ cell stages during the cycle of the seminiferous epithelium is extremely difficult in most mammalian species due to the continual presence of different spermatogenetic stages in the testis from the onset of puberty. The problem is also compounded by the uniform distribution of both seminiferous tubules and interstitial tissue throughout the entire testis. This difficulty can be circumvented, however, by studying certain species where there is a topographical distribution of germ cell stages within the testis. The urodele amphibian Necturus maculosus exhibits a breeding cycle during which a longitudinal wave of spermatogenesis occurs along the length of the testis, resulting in a spatial and temporal segregation of differentiating germ cells. Moreover, this topographical pattern of spermatogenesis is also reflected in the degree of development of adjacent Leydig cells. This anatomical arrangement allows distinct testicular regions to be obtained using a dissecting microscope. The isolated zones, containing germ cells and Leydig cells in various stages of development, were analyzed for 17 alpha-hydroxylase, C-17,20-lyase, and aromatase activities (key enzymes for the synthesis of androgens and estrogen), estrogen binding, and cytochrome P-450 content. Functional parameters were then correlated with the morphology of Leydig cells in the various zones observed by both light and electron microscopy. It was found that there existed a distinct correlation between the state of differentiation of the leydig cells, their steroidogenic potential, and the distribution of estrogen receptors. These results in Necturus indicate indicate in this species, at least, the steroidal microenvironment of different germ cell associations may be quite specific.     

Infertility and testicular defects in hormone-sensitive lipase-deficient mice

The 84-kDa hormone-sensitive lipase (gene designation Lipe; EC 3.1.1.3) is a cholesterol esterase and triglyceride hydrolase that functions in the release of fatty acids from adipocytes. The role of hormone-sensitive lipase in other tissues such as the testis, where a specific 120-kDa testis-specific isoform is expressed, is unknown. To study this, we examined the fertility and testicular histology of gene-targeted hormone-sensitive lipase-deficient mice. Homozygous hormone-sensitive lipase-deficient male mice are infertile and have decreased testis weights; female homozygotes are fertile. Testicular abnormalities, detected at the light and electron microscopic levels, included the presence of multinucleated round and elongating spermatids, vacuolization of the seminiferous epithelium, asynchronization of the spermatogenic cycle, sloughing of postmeiotic germ cells from the seminiferous epithelium into the lumen, and a marked reduction in the numbers of late spermatids. Extensive nuclear head deformation was noted in late spermatids as well as the sharing of a common acrosome in multinucleated cells. In some multinucleated cells, nuclei were separated from their acrosomes, with the acrosomes remaining attached to areas of ectoplasmic specializations, suggesting defects in intercellular cytoplasmic bridge integrity. Although the lumen of the epididymis was essentially devoid of spermatozoa and filled instead with spherical degenerating cells, the epididymal epithelial cells appeared normal. The few late spermatids present in the epididymis were abnormal. There was no morphological evidence, as judged by the absence of lipid droplets of triacylglycerol or cholesteryl ester accumulation in the testis. Together, the data suggest that hormone-sensitive lipase deficiency results in abnormalities in spermiogenesis that are incompatible with normal fertility. We speculate that a metabolite downstream from the hormone-sensitive lipase reaction may be essential for membrane stabilization and integrity in the seminiferous epithelium and, in particular, may play an important role in the maintenance of intercellular cytoplasmic bridges between postmeiotic germ cells.     

Fine structure of the Sertoli cell of the testis of the teleost Oryzias latipes

The epithelium lining spermatogenic cysts in the testis of the teleost Oryzias latipes was investigated with the electron microscope. The cyst epithelium (CE) is simple squamous until the onset of the transformation of spermatids into spermatozoa, at which time the CE become columnar. The squamous CE cell is relatively poor in organelles, but the columnar CE cell contains numerous glycogen granules and smooth endoplasmic reticulum, and shows evidence of much phagocytosis. A limiting membrane in the subjacent connective tissue is described.The cytological features and histological relationships of the CE are compared with those of the Sertoli cell of the mammalian seminiferous epithelium. The CE cell is believed to be the homolog of the Sertoli cell in testes of teleosts.     

金雀异黄素对良性前列腺增生小鼠血清睾酮水平的影响

目的 观察金雀异黄素（genistein，Gen）对小鼠良性前列腺增生（BPH）的抑制作用及对血清睾酮水平的影响。方法 激素法诱发小鼠BPH，称重法测定前列腺湿重，计算前列腺指数；光镜和电镜下观察前列腺组织形态及超激结构的变化，并进行形态定量学分析；放免法测定血清睾酮水平。结果 制模1w后Gen治疗3w，20、40、160mg／kg均可剂量依赖性降低BPH小鼠前列腺湿重指数。光镜下见增生的腺上皮乳头减少或消失，腺体上皮细髓呈低立方或扁平状，腺腔明显扩张或萎缩；图像分析显示，Gen明显减小BPH小鼠前列腺腺体平均最小直径、最大直径及平均面积均减小（P〈0.05）。电镜下，Gen治疗组腺上皮细胞排列规整，呈低柱状，核位于基底部，核仁多为一个，线粒体数量不多，粗面内质网较多，高尔基复合体较发达，分泌颗粒减少。有微绒毛脱落和核膜完整的腺上皮细胞核脱落到腺腔的现象。结论 Gen可明显抑制小鼠BPH，降低BPH小鼠血清睾酮水平，提示其抗BPH作用机制可能涉及抗雄激素作用。