雷公藤单体(TW_(19))雄性抗生育活性的研究

雷公藤单体(TW_(19))系从雷公藤中分离、提纯出的一个二萜类化合物,本实验观察其对雄性大鼠抗生育作用及可能机理.TW_(19)(400μg/kg·d)给药三周后无抗生育作用,五周后生育力丧失,附睾尾部精子活力和密度明显降低,畸形率升高,附睾重量减轻,但对附睾上皮的形态变化无明显影响,管腔内可见畸形精子及脱落生精细胞.附睾尾部液中肉毒碱含量,给药组较对照组显著降低.TW_(19)对附睾头部α-糖苷酶和酸性磷酸酶含量没有明显影响;对生精上皮影响轻微,各期曲细精管基本正常,少数表现出精子细胞轻度损伤;TW_(19)可使睾丸重量减轻,并可显著降低睾丸透明质酸酶含量,但对睾丸乳酸脱氢酶-C_4没有明显抑制作用;对前列腺、储精囊等附属性腺器官重量没有影响.结论:TW_(19)对雄性大鼠具有确定的抗生育效果,作用环节可能在精子细胞及附睾.     

Antifertility effects of alcoholic seed extract of Abrus precatorius Linn. in male albino rats

Effects of alcoholic seed extract of Abrus precatorius Linn. were investigated at a dose of 100 mg/Kg body wt./day/rat for 60 days on fertility, semen profile and sperm metabolism of orally administered sexually mature male albino rats using WHO protocols. Serum testosterone levels were also measured using RIA technique. The data revealed that the cauda epididymal sperm motility was significantly lowered with no effect in its sperm concentration by 60 days of feeding. The scanning electron microscopic study on sperm morphology exhibited decapitation, acrosomal damage and formation of bulges on midpiece region of sperms in treated rats. The biochemical studies on epididymal spermatozoa indicated alterations in their energy and/or oxidative metabolism as evidenced by a fall in succinate dehydrogenase and ATPase levels by extract allocation. It did not exert any effect in body and organ weights. But an average number of implantation sites in females after mating with the treated male rats markedly declined. Contrarily, a significant increase in serum testosterone levels was noted by 60 days of administration. Thus, the decrease in fertility rate in extract receiving animals is correlated with reduced sperm motility, metabolism and altered sperm morphology in epididymis.     

Comprehensive assessment of the effect of Sphenocentrum jollyanum root extract on male reproductive activity in albino rats

Aim  To evaluate the effect of methanol extract from the Sphenocentrum jollyanum root on male reproductive activity. Methods  Male albino rats were treated orally with distilled water (vehicle for the extract; control) and 50, 100 and 150 mg kg^-1 body weight of Sphenocentrum jollyanum root extract for 8 weeks. Each group had its own recovery. Rats were killed 24 h after the last treatment. Caudal epididymal sperm count, motility, viability, morphology and organ weights were determined. Hematological indices, serum proteins, enzymes, testicular Superoxide dismutase (SOD) activity, and testicular and epididymal histology were determined. Results  Compared with the control, the extract caused a dose dependent significant (P< 0.05) reduction in progressive motility of spermatozoa, viability and total sperm counts. The number of abnormal spermatozoa and epididymal volume were not statistically significant. There was a significant increase (P< 0.05) in serum testosterone levels in rats treated with 50 (P< 0.01) and l00 mg kg⁻¹ (P< 0.05) of Sphenocentrum jollyanum. There was a significant (P< 0.05) increase in red blood cell count, packed cell volume and hemoglobin concentration, whereas there was no change in white blood cell count, mean total serum protein, albumin and globulin in the sera of Sphenocentrum jollyanum treated rats when compared with the control. The extract caused a significant decrease (P< 0.05) in serum aspartate and alanine aminotransferase activities with a significant increase (P < 0.05) in testicular SOD activity at a dose of 50 mg kg⁻¹ bodyweight. Testicular cytoarchitecture of the extract treated rats showed degeneration of seminiferous tubules, whereas regeneration of germinal epithelium and restructuring of the germinal interstitium occurred in the recovery rats. No lesions were observed in the epididymis of the rats. Conclusion  The results suggest that methanol extract of the Sphenocentrum jollyanum root could produce harmful effects on reproductive functions in male albino rats which can be attributed to poor sperm quantity (epididymal sperm count), quality (sperm motility, viability and morphology) and testicular degeneration. The steroidogenic potential of the plant could explain its use as an aphrodisiac agent.     

不育症患者精子X,Y及18染色体的荧光原位杂交分析

目的:分析不育症患者精子X,Y,18染色体的荧光原位杂交情况。方法:在男性不育症组中,2例无精子症患者从附睾抽吸获取精子、3例从睾丸获取精子、2例严重少精症患者从射出精液中找到精子。选择5例正常射出精液者作为对照组。应用荧光原位杂交法(FISH)检查精子X,Y以及18号染色体,比较两组精子染色体非整倍体的发生率。结果:不育症组睾丸精子、附睾精子的非整倍体率无差别,但不育症组精子与正常男性组精子比较,非整倍体总发生率、性染色体二体性率及缺对染色体率明显增高(2.8％vs0.58％,0.81％vs 0.19％,2.1％vs0.37％),P<0.001。结论:无精子症与严重少精子症患者的精子比正常精子具有更高的染色体非整倍体率,需要进行大样本的研究,为不育症患者的治疗和遗传咨询提供有效的证据。     

Human male infertility caused by degeneration and death of sperm in the epididymis

Four patients with persistent oligospermia and necrospermia were found to have severely degenerated sperm in the ejaculate. However, in those examined, testicular sperm were ultrastructurally normal, indicating that sperm degeneration and death was occurring during epididymal passage or storage or both or upon mixing with the seminal plasma at ejaculation. Seminal plasma was found to be nontoxic to normal donor sperm. In three patients, frequent ejaculation (two ejaculates per day for 4 or 5 days) was used to deplete epididymal sperm reserves and reduce the period spent in the epididymis. This resulted in a threefold to sevenfold increase in percentage of motile sperm in the ejaculate and a similar increase in sperm motility index. The authors propose the term "epididymal necrospermia" to describe this previously undefined type of male infertility.     

大鼠储精囊分泌蛋白Ⅱ在精子表面的荧光定位研究

本文报道应用免疫荧光定位技术首次证明大鼠储精囊分泌蛋白Ⅱ(SVPⅡ)和附睾精子表面结合。纯化的SVPⅡ和附睾精子一起培育后,经兔抗SVPⅡ抗血清和FITC标记的羊抗兔抗体共同保温,在荧光显微镜下观察,发现整个附睾精子表面被黄色的荧光所覆盖,且以精子头部为著;而同时以兔免疫前血清取代兔抗SVPⅡ抗血清作对照,精子表面未见任何荧光结合区。实验结果充分表明:SVPⅡ能和附睾精子表面呈特异结合,可能为大鼠精子包被抗原的成分。另外经免疫吸收试验和免疫交叉反应提示:SVPⅡ为组织特异抗原,与已纯化的其它SVP组分无交叉反应。     

A factor in human seminal plasma which affects carnitine accumulation in bovine epididymal sperm

This study was initiated to determine whether factors are present in human sperm-free seminal plasma (HSP) that regulate the uptake and release of carnitine from sperm. Bovine caput epididymal sperm cells accumulated more carnitine than caudal sperm cells. A significant reduction in carnitine uptake by caput sperm was observed in the presence of HSP from normal subjects, but not from three subjects with reduced motility. A factor has been isolated from HSP that inhibits carnitine uptake by caput sperm and has the following properties: it is nondialyzable, stable to freeze-thawing, soluble in 60% ammonium sulfate, and has an approximate molecular weight of 158 kd. These data are consistent with the existence of a relatively high molecular weight protein in HSP responsible for the preservation of carnitine concentrations in sperm.     

Improved seminal characteristics in infertile men after a conservative treatment regimen based on the avoidance of testicular hyperthermia

Evidence to support the contention that avoidance of testicular hyperthermia (due to hot baths and/or tight clothing) will improve fertility is, in the main, anecdotal. Semen samples derived from 128 infertile men were analyzed before and after a 3-month conservative treatment regimen. Six seminal characteristics were studied. Clinical evaluation included normal bath temperature, types of underwear worn, and past history of hernia repair, orchiopexy, or varicocelectomy. The patients were divided into three groups (A to C) according to sperm density and then subclassified on the basis of progressive motility status. The results show that a significant, or approaching significant, improvement in seminal characteristics occurs in certain groups of oligospermic individuals with pretreatment progressive motility of less than 40%. In clinical trials, it should be recognized that any improvement in seminal quality following pharmacologic or surgical intervention may include a contribution from conservative treatment.     

Further studies on the fertility promoting factor from human seminal plasma.

A small molecular weight substance from human seminal plasma has been further purified by chromatography. The fertility promoting action of this factor on epididymal sperm has been confirmed in mouse in vitro and in vivo. Experimental evidence indicates that the factor acts on the sperm and not on the eggs. Its possible mode of action is by improving the motility and survival of the epididymal sperm.     

大鼠附睾精子-SH变化与运动关系的研究

本文以巯基(-SH)分子探针mBBr标记附睾精子,采用流式细胞仪分析,结合荧光显微镜观察;以研究附睾精子膜成熟变化,其结果显示:精子在附睾运行过程中,-SH渐渐氧化为二硫键(-S-S-)、并具异质性.Percol法筛选运动精子的研究表明:运动精子的-SH含量显著降低,这提示附睾精子膜-SH与-S-S的变化与精子运动的获得和发展有关.     

Altered structural and functional integrity of the reproductive tissues in estradiol benzoate-treated intact male albino rats

The effects of estradiol benzoate administration (50 micrograms/day/rat for 15 days) to healthy intact male albino rats of Holtzman strain were investigated. Estradiol benzoate (E2B) treatment caused significant reduction in androgen-target tissues, viz., testis, epididymis, vas deferens, seminal vesicle, and ventral prostate. This led to alterations in the milieu of the epididymis and vas deferens rendering them hostile for sperm maturation, motility, metabolism, and survival as is evident by reduced sperm motility, alterations in their morphology, and the resulting loss of fertility. The reduction in sperm counts are correlated with decrease in weight, atrophy of testis, and arrest of spermatogenesis. It is evident that E2B treatment to adult intact, male rats manifested antiandrogenic and antifertility effects mainly by directly affecting the testicular structural and functional integrity in support of earlier data. Studies on induction of functional sterility after E2B treatment are called for.     

Adrenomedullin in human male reproductive system

OBJECTIVE: To investigate adrenomedullin (AM) localization and distribution in human male reproductive system and to determine whether seminal fluid AM concentration correlates with sperm parameters. STUDY DESIGN: Plasma and semen samples (n = 19) obtained from healthy volunteers with normal seminal fluid parameters were assayed for AM using a specific RIA. AM immunostaining was sought on sections of penile cavernous bodies and testicular tissues obtained postmortem from four young males after accidental death using a polyclonal antibody to AM 1-52. RESULTS: Mean AM concentration in seminal plasma was 209.4+/-46.6 pg/ml, 8-9-fold higher than in circulating plasma and correlated with sperm motility (r = 0.715, p < 0.01). Endothelial cells of cavernous vessels stained for AM. Intense AM immunostaining was found in germinal cells and in peritubular myocytes and Leydig cells in the testis. CONCLUSIONS: These findings demonstrated for the first time that AM is localized in human male reproductive system. The local secretion of AM suggests that AM may contribute either in the penile erection and in the regulation of testicular function and sperm motility.     

青春期大鼠受己烯雌酚持续作用对成年后睾丸结构与功能的影响

目的：探讨青春期大鼠受己烯雌酚(DES)持继作用后的睾丸损伤,损伤后在成年期的睾丸结构及生精功能能否恢复。方法：青春期雄性SD大鼠48只,随机均分成：A_1、B_1、C_1和D_1组；A_2、B_2、C_2和D_2组。分别隔日腹腔注射含DES0μg·kg~(-1)·d~(-1)(A_1,A_2,为对照组)、2μg·kg~(-1)·d~(-1)(B_1,B_2)、10μg·kg~(-1)·d~(-1)(C_1,C_2)和50μg·kg~(-1)·d~(-1)(D_1,D_2)的玉米油0.3 ml,28 d后处死A_1-D_1组动物采集标本。A_2-D_2组则继续正常饲养60 d(恢复期),然后处死动物取材。检测大鼠体重(BW)、睾丸重量(TW)、附睾重量(EW)、睾丸每日精子生成量(DSP)以及附睾尾精子数(ESC)等5项参数的变化,应用光镜观察睾丸生精上皮的组织学变化。结果：B_1组的TW、DSP和ESC显著低于A_1组(P＜0．01),C_1组和 D_1组的5项参数均显著低于A_1组(P＜0．05)。与A_2组相比较,B_2组的5项参数均没有显著性差异(P＞0．05),C_2组DSP和ESC显著减少(P＜0．05),D_2组则TW、EW、DSP和ESC显著降低(P＜0．01)。DES染毒各组的睾丸曲细精管出现了不同程度的萎缩,且随剂量增加病理改变程度加重。恢复期后,睾丸曲细精管组织结构B_2组显示正常,C_2组有明显恢复,D_2组未得到恢复。结论：青春期大鼠受DES持续作用可导致睾丸生精上皮损伤,生精能力下降。损伤较轻组在成年后,睾丸组织结构和生精功能得到不同程度的恢复,但损伤程度重者未见恢复。     

Semen analysis results and air pollution in the group of men from infertile couples in the Lower Silesia in 1977-2000

OBJECTIVES: To evaluate the relation among the concentration of selected air pollution and seminal parameters, examined from 1977 to 2000. DESIGN: Semen analysis and air pollution results were retrospectively evaluated. MATERIALS AND METHODS: We have analysed semiograms from 1363 men from infertile couples inhabiting Lower Silesia. Seminal volume, sperm concentration, percentage of pathologic sperms were measured in all men. Estimation of mean seminal volume, total sperm number, sperm motility, and percentage of pathologic sperms per year was performed. Average values for NO2, CO, SO2 and dust concentrations during the study were taken from official sources. RESULTS AND CONCLUSIONS: We showed the statistically significant increase in the percentage of pathological sperms (R2 = 0.9, p < 0.05), the slight increase in the semen volume(R2 = 0.4, p < 0.05). The total sperm count in the semen samples revealed a slight increase (R2 = 0.25, p < 0.05). The sperm concentrations and the percentage of motile sperms remained relatively stable and relatively constant. Statistically important decrease in NO2 (R2 = 0.85, p < 0.05), SO2 (R2 = 0.96, p < 0.05) and dust concentration (R2 = 0.89, p < 0.05) and no change in CO concentration was revealed. There is no correlation among concentrations of dust, NO2, SO2, CO and the increase in percentage of pathologic sperms.     

Severe oligoasthenoteratozoospermias,secretory and obstructive azoospermias: Motility as a criterion of sperm viability

Purpose: Comparison of pregnancy rates in cases of Secretory Azoospermias (SA), Obstructive Azoospermias (OA) and severe Oligoasthenoteratozoospermias (OATZ). Evaluation of sperm motility as a quality criterion. Methods: In SA cases (n = 35), 9 samples were cryopreserved. In OA cases (epididymal aspiration: n = 91; testicular biopsy: n = 206), all samples were cryopreserved. 596 OATZ ejaculates were included. Results: In SA cases, 2 pregnancies were achieved from 9 ICSI cycles. In OA, motile sperm rates were higher in testicular biopsies. After thawing sperm motility was not different between testicular and epididymal origin. 2 pregnancies were achieved with immotile testicular sperm after thawing, but none with immotile epididymal sperm. In OATZ cases, one pregnancy was obtained from the 9 cryopreserved ejaculates and 35.3% with fresh motile sperm. Conclusions: In SA cases, the use of donor sperm is recommended due to the lower pregnancy rate achieved. Motility, before and after cryopreservation, as a criterion of sperm viability is discussed and its use should be reconsidered in some cases.     

Participation of vitamin A in the maturation of rabbit spermatozoa.

Vitamin A concentration was fluorometrically measured in epididymal and ejaculated rabbit spermatozoa and in some of the sperm cells subcellular components. The concentration of vitamin A in the epididymal cells was about one-half that observed in the ejaculated spermatozoa (2.68 as against 1.05 mug/10(8) cells) and seemed to be the same in the sperms obtained from both the head and the tail of the epididymus. The concentration of vitamin A was also found to be significantly higher in the seminal plasma than in the epididymal secretion (0.06 as against 0.039 mug/mg protein respectively). Practically all the vitamin A was found in the fractions obtained by treatment with hypotonic MgCl2 (acrosomal region) and/or with hyamine and dithiothreitol (plasma membrane). It was concluded that the sudden increase in the sperm concentration of vitamin A that occurs upon ejaculation may be required for the stabilization of the acrosomal and plasma membranes.     

少量人类精子以空卵透明带为冻贮载体的冷冻保存

目的:探讨空卵透明带作贮存载体冷冻保存少量人类精子。方法:将人或金黄地鼠卵内的细胞成份全部除去,制备成空卵透明带,分别显微注入人睾丸精子、附睾精子和射出精子后冷冻保存,并与正常供精者射出精子作对照。结果:解冻后卵透明带在溶液中容易识别寻找,卵透明带内的精子易于观察。附睾精子组(n=11)与供精者射出精子组(n=7)的冷冻精子活动率和存活率无显著性差异(P>0.05),但睾丸精子组(n=7)这两项参数值均显著低于附睾精子和射出精子组(P<0.01)。含6％、7.5％和9％不同甘油浓度的冷冻保护剂,对冷冻精子活动率无显著性影响(P>0.05);冻贮于人和金黄地鼠空卵透明带内的精子,两者的冷冻精子活动率也无显著性差异(P>0.05〕。结论:空卵透明带是冻贮少量人类精子的合适载体。     

On the maintenance of male fertility in the absence of native testosterone secretion: site-directed hormonal therapy in the rat

A method of direct percutaneous injection of testosterone (T)-laden microspheres directly into the testis was used in an attempt to achieve the maintenance of normal intratesticular T concentrations, spermatogenesis, and fertility. Rats were divided into three groups: (1) sham operated/injection controls; (2) animals receiving 250 micrograms/d gonadotropin-releasing hormone (GnRH)-antagonist; and (3) animals receiving GnRH-antagonist as in group 1 plus 20 mg T-laden microspheres/testis. Treatment periods were 45 and 90 days. Serum T, testicular interstitial fluid T, testis weights, epididymal weights, daily sperm production (sperm x 10(6)/g/d), cauda sperm motility, and fertility were assessed in all animals. Gonadotropin-releasing hormone antagonist treatment reduced serum and testicular interstitial fluid to below detectable levels at day 45 and to similar levels at day 90. Supplementation with T-laden microspheres maintained testicular interstitial fluid T at concentrations not different from controls without elevation of serum T concentrations. All other values, including fertility were suppressed by GnRH-antagonist treatment and maintained by supplementation with T-laden microspheres.     

Einsatz der Kryotechnologie in der Behandlung männlicher Infertilität

The cryopreservation technique has a great impact in the treatment of male infertility. Especially in males undergoing chemotherapy for oncological diseases, cryopreservation of sperm samples may help to preserve the chance to father a child after completion of the oncological treatment. Furthermore, cryopreservation is necessary to optimize the screening of sperm donors for insemination. The cryopreservation of testicular samples or epididymal aspirates in patients suffering from azoospermia helps to avoid unnecessary hormonal stimulation procedures in the female partner. Only in cases in which fertilizable sperms are present should an intracytoplasmic sperm injection cycle be planned. The success rates of treatment using frozen or non-frozen sperm samples seem to be the same, especially since the introduction of intracytoplasmic sperm injection. This helps to overcome the lower fertilizing capacity of sperms following cryopreservation due to loss of quality.     

氯化镉对大鼠附睾结构及精子的影响

目的:探讨氯化镉(CdCl2)对大鼠附睾组织学和附睾精子参数的影响。方法:20只雄性SD大鼠随机均分为对照组与3个实验组,每组5只动物。实验组分别腹腔注射CdCl2 0.2mg/kg(低剂量组),0.4mg/kg(中剂量组)和0.8mg/kg(高剂量组)×14d,对照组注射等量生理盐水。观察附睾上皮的组织学改变和附睾精子参数变化。结果:低剂量组的附睾上皮未见明显组织学病理改变。随着Cd2+剂量增加,实验组附睾主细胞的胞质疏松明显,胞内空泡化程度加大,核染色质边聚,静纤毛缩短,附睾管腔内的精子明显减少,多见降解的生精细胞。3个剂量组的精子存活率、精子活动率、精子浓度和正常形态精子率均显著低于对照组(P<0.01)。结论:CdCl2染毒可导致大鼠附睾组织学损伤性改变,附睾精子参数显著降低,且损伤呈剂量效应。