非梗阻性无精子症患者的表皮生长因子及受体表达

目的:探讨血清表皮生长因子(EGF)和睾丸组织表皮生长因子受体(EGF-R)与非梗阻性无精子症睾丸生精功能障碍的关系。方法:采用放免法对20例无精子症患者和20例已育男性志愿者进行血清EGF测定;无精子症患者接受睾丸活检,病理检查按Johnson评分法评价睾丸精子发生及障碍的程度;采用免疫组化SABC法对睾丸3种主要细胞进行EGF-R表达的检测。结果:无精子症患者血清EGF浓度明显低于男性对照者(P<0.05)。睾丸活检生精功能评为8分者14例;3分者2例;6、5、4和 2分者各1例。在20例无精子症睾丸的间质细胞、支持细胞和生精细胞均有EGF-R的表达,间质细胞EGF-R强阳性和阳性表达明显低于支持细胞和生精细胞(P<0.05)。结论:无精子症患者血清EGF浓度下降、间质细胞EGF-R强阳性和阳性表达的减少与睾丸生精功能障碍的程度是相一致的,说明血清EGF及睾丸EGF-R在调节睾丸生精功能方面有重要的作用。     

短期糖尿病大鼠睾丸的细胞学变化

目的:探讨糖尿病发病初期大鼠发生性与生殖功能障碍的细胞学机制。方法:通过给成龄雄性大鼠腹腔注射链脲菌素制成糖尿病动物模型,分别观察1d、3d、5d、7d、14d。观察期满后断头处死,取睾丸。应用常规组织学方法观察睾丸组织形态的改变;应用流式细胞分析术检测睾丸间质细胞、支持细胞及各级生精细胞的细胞周期的改变。结果:睾丸间质细胞及生精细胞的形态、数目从致病d7开始有明显变化,支持细胞的形态无明显变化;睾丸中各类细胞在高糖环境下24h即发生增殖减慢、细胞滞留在G1期、DNA合成减少等改变。结论:糖尿病大鼠生殖功能障碍与睾丸间质细胞、生精细胞的形态、数目变化及睾丸各类细胞的增殖改变有关。     

环境剂量的双酚A暴露对雄性小鼠生殖功能的影响

目的:研究环境剂量的双酚A(BPA)暴露对雄性小鼠生殖功能的影响及可能的分子机制。方法:84只3周龄C57BL/6J雄性小鼠按体质量随机分为3组,分别为5μg/kg BPA组、50μg/kg BPA组、乙醇溶剂对照组,每组28只。每日灌胃给药1次,连续给药5周后,检测生殖器官脏器指数、附睾尾精子数量、睾丸组织学形态;同期与正常雌性小鼠配种检测雄鼠的生育力。应用MBDq PCR方法检测睾丸中Tnnt2、Tectb基因的甲基化水平变化。结果:1 BPA暴露组与对照组间的生殖系统脏器指数无统计学差异(P0.05)。2 50μg/kg BPA暴露组附睾尾精子数量下降20.1%;睾丸曲细精管管腔中有生精细胞脱落等病理现象。3 BPA暴露组与对照组间的生育率无统计学差异(P0.05),但50μg/kg BPA暴露组每胎平均产仔数下降。4 BPA组睾丸中Tnnt2、Tectb基因的甲基化水平下降(P0.01)。结论:环境剂量的BPA暴露对雄鼠生殖器官脏器指数没有影响,50μg/kg BPA能引起小鼠生精功能下降,睾丸Tnnt2、Tectb基因的甲基化水平下降。     

恒猴精索内注射长效局麻药后对睾丸和附睾的影响

在13只恒河猴精索内注射长效局麻药后,睾丸发生明显变化;而且给药剂量不同,睾丸曲精小管改变的程度和恢复正常的时间亦不同。睾丸间质细胞无明显变化;睾酮分泌不减少,无毒性反应。对临床男性节育应用及生殖生理研究均具有意义。     

铅离子对雄(男)性生殖系统的毒性影响

铅离子(Pb2+)对雄(男)性生殖系统具有毒性作用。Pb2+可致睾丸、附睾及其附属性腺重量下降,生精上皮损伤,各级生精细胞脱落,支持细胞和间质细胞退化,睾酮水平下降,参与睾酮合成的酶活性被阻断等功能异常。精子发生减少、畸形精子增多、精子运动能力下降、活性氧增多等,引致雄(男)性生育力下降。本文综述Pb2+对睾丸、附睾及附属性腺的影响及其作用机制。     

脂多糖诱导的非特异性炎症对大鼠睾丸功能的影响

目的:探讨脂多糖(LPS)诱导的非特异性炎症对大鼠睾丸功能的影响。方法:利用磁分离均相酶联免疫技术、免疫组织化学、原位杂交方法分别检测血清睾酮(T)、黄体生成素(LH)水平以及生精上皮内增殖细胞核抗原(PCNA)、雄激素结合蛋白(ABP)mRNA的表达。结果:与对照组相比实验组血清T升高、ABPmRNA表达增多(P<0.05)以及生精上皮PCNA表达明显减低(P<0.05),而血清LH变化无显著差异(P>0.05)。结论:睾丸炎症时间质细胞、支持细胞功能改变,致使睾丸局部生精微环境发生变化,可能导致生精障碍。     

昆明山海棠片所致大鼠睾丸损伤的病理学研究

目的:观察昆明山海棠片连续给药90 d对大鼠睾丸结构的损伤。方法:80只SD雄性大鼠随机分为5.0 g/kg、10.0 g/kg、20.0 g/kg 3个给药组及正常对照组(等量蒸馏水),连续灌胃3个月。于给药1个月、2个月、3个月,采用光学显微镜、透射电子显微镜观察睾丸组织形态学的变化。结果:各给药组均可引起大鼠生精细胞损伤和破坏;睾丸超微结构的改变主要包括各级生精细胞坏死、空泡化,细胞质内溶酶体数量增多,线粒体肿胀、固缩,内质网扩张,细胞核染色质异常分布、溶解、固缩,核崩解消失等;支持细胞、间质细胞未见明显异常。结论:昆明山海棠片连续给药主要导致大鼠睾丸生精细胞超微结构损伤,其损伤存在剂量效应和时间效应关系。     

脉冲式注射LHRH诱发特发性低促性腺激素型性功能低下患者的精子发生

本文首次在国内用自制YED-1型自动注射泵脉冲式皮下注射LHRH(5μg/脉冲/90分)治疗4例低促性腺激素型性功能低下(IHH)。所有4例经治疗后症状明显改善,性激素水平明显提高,睾丸体积增大,其中2例达正常成年男子水平,并在治疗的第250和330天时精液中出现成熟精子,精子计数分别为4×10~6/ml和10×10~6/ml,第3例睾丸由治前的4ml增大到8ml。2例已结婚。第4例经治疗4个月,症状及激素水平明显好转后,出现耐药现象,血清T水平又下降到治疗前的极低水平,阴茎停止勃起,推测可能与出现抗LHRH抗体有关。     

超声波对家兔抗生育效应研究

一定剂量的超声波,对家兔睾丸有抑制生精作用。使用1.5W/cm~2超声功率,可迅速抑制睾丸生精作用。超声波对动物睾丸无刺激、无损伤,对内分泌及转氨酶均无影响。停止超声作用仍可恢复生育能力,所产子兔无畸形。本文对超声波抗生育机制进行了初步探讨。     

长期使用十一酸睾酮抑制大鼠睾丸和附睾的雄激素受体基因表达

本研究探讨十一酸睾酮抗雄性大鼠生育的机制。１２周龄雄性ＳＤ大鼠,给药组８只每２周注射２０ｍｇ／ｋｇ十一酸睾酮,共３个月。与１０只对照大鼠比较,十一酸睾酮处理大鼠睾丸网液中精子密度下降到对照组的７％,附睾尾部精子活动力下降至对照组的６％,血清睾酮水平上升至对照组的２５５％,而睾丸网液睾酮却下降至对照组的５５％。两组之间均有非常显著差异。未发现十一酸睾酮处理大鼠睾丸生殖细胞和附睾上皮细胞凋亡状况有明显改变,但睾丸和附睾的雄激素受体基因表达显著低于对照组大鼠。十一酸睾酮抑制睾丸精子生成和降低附睾精子活动力可能与睾丸网液的雄激素水平下降以及睾丸和附睾雄激素受体基因表达的抑制有关。     

Relationship of leptin administration with production of reactive oxygen species, sperm DNA fragmentation, sperm parameters and hormone profile in the adult rat

Purpose

Leptin, an adipose tissue-derived hormone, plays an important role in energy homeostasis and metabolism, and in the neuroendocrine and reproductive systems. The function of leptin in male reproduction is unclear; however, it is known to affect sex hormones, sperm motility and its parameters. Leptin induces mitochondrial superoxide production in aortic endothelia and may increase oxidative stress and abnormal sperm production in leptin-treated rats. This study aims to evaluate whether exogenous leptin affects sperm parameters, hormone profiles, and the production of reactive oxygen species (ROS) in adult rats.

Methods

A total of 65 Sprague–Dawley rats were divided into three treated groups and a control group. Treated rats received daily intraperitoneal injections of 5, 10 and 30 μg/kg of leptin administered for a duration of 7, 15, and 42 days. Control rats were given 0.1 mL of 0.9 % normal saline for the same period. One day after final drug administration, we evaluated serum specimens for follicle-stimulating hormone (FSH), leutinizing hormone (LH), free testosterone (FT), and total testosterone (TT) levels. Samples from the rat epididymis were also evaluated for sperm parameters and motility characteristics by a Computer-Aided Semen Analysis (CASA) system. Samples were treated with 2′,7′-dichlorofluorescein-diacetate (DCFH-DA) and analyzed using flow cytometry and TUNEL to determine the impact of leptin administration on sperm DNA fragmentation.

Results

According to CASA, significant differences in all sperm parameters in leptin-treated rats and their age-matched controls were detected, except for TM, ALH and BCF. Serum FSH and LH levels were significantly higher in rats that received 10 and 30 μg/kg of leptin compared to those treated with 5 μg/kg of leptin in the same group and control rats (P < 0.05). ROS and sperm DNA fragmentation was significantly higher in rats injected with 10 and 30 μg/kg of leptin for 7 and 15 days compared with rats treated with 5 μg/kg of leptin and the control group (P < 0.05) for the same time period. However, at day 42 of treatment, ROS and sperm DNA fragmentation levels significantly decreased in all groups (P < 0.05).

Conclusion

According to these results, leptin can possibly affect male infertility by ROS induction or hormone profile modulation.     

The prostaglandin inhibitor effect of antiinflammatory drugs in the therapy of male infertility

The effects of various doses of indomethacin and ketoprofen as compared with placebo were examined in 100 oligospermic patients who participated in this study. It was found that the treatment increased sperm count, sperm motility, and fertilizing capacity. The radioimmunoassay examination showed an increase in plasma follicle-stimulating hormone and luteinizing hormone but a decrease in testosterone. The results show that the influence of indomethacin was better in the dose of 75 mg daily. The mechanism of those changes is not clear. Treatment should be at least 60 days.     

Does constant infusion of gonadotropin-releasing hormone agonist lead to greater suppression of gonadal function in man than its intermittent administration?

Constant infusion of gonadotropin-releasing hormone agonist (GnRH-A) in the rhesus monkey leads to far greater suppression of spermatogenesis than its intermittent administration. To assess whether administration of GnRH-A by constant subcutaneous infusion will also lead to greater inhibition of gonadal function, we administered either 20 or 200 micrograms of D(Nal2)6GnRH (GnRH-A) to two groups of normal male volunteers, either by a single daily injection or constant subcutaneous infusion through a portable infusion pump, for 28 days. Basal and integrated luteinizing hormone (LH), follicle-stimulating hormone, and testosterone (T) responses were not significantly different between the two methods of GnRH-A administration at either the 20- or 200-microgram dose, even though subjects in the constant infusion group showed more consistent inhibition of basal and 24-hour integrated T concentrations. Significant decline in serum T in both groups occurred in the face of little or no decline in serum LH. It is concluded that the effects of constant infusion of GnRH-A in man are not as striking as those reported in the rhesus monkey and that the antigonadal effects of GnRH-A in man are complex and cannot be explained on the basis of down-regulation of pituitary gonadotropin secretion alone--additional mechanisms may be operative.     

Active immunization with follicle-stimulating hormone for fertility control: a 4 1/2-year study in male rhesus monkeys

Active immunization of four adult rhesus monkeys with highly purified ovine follicle-stimulating hormone (FSH) over 4 1/2 years resulted in the production of specific FSH antibodies. While luteinizing hormone and testosterone secretion were not affected, sperm counts were in most instances reduced to or below the lower normal range. On a few occasions, azoospermia or high sperm counts were observed. Although the antibodies produced neutralized the biologic activity of FSH throughout, spermatogenesis gradually returned, as evidenced by testicular histologic characteristics. However, the diameter of the seminiferous tubules remained decreased, and the germinal epithelium was depleted. No adverse side effects could be demonstrated in the immunized animals, e.g., immune complexes in either serum or tissues and resultant tissue damage. These results show that although active immunization with FSH may not result in an effective method of male fertility control, long-term immunization against a circulating hormone may not result in deleterious side effects.     

Effect of follicle-stimulating hormone upon estrogen and progesterone secretion by cultures of monkey granulosa cells recovered during the periovulatory period

For determination of how exposure of the monkey follicle to the preovulatory luteinizing hormone (LH)/follicle-stimulating hormone (FSH) surge alters its responsiveness to FSH in terms of estrogen and progesterone secretory ability, monkey thecal tissue and granulosa cells were harvested prior to and during the midcycle LH/FSH surge and cultured for 8 days with testosterone and with and without 100 ng human FSH. The addition of FSH enhanced estrogen secretion in culture (6.8 and 7 times on the average after 6 and 8 days, respectively; P less than 0.05) by granulosa cells if they were harvested prior to, but not during, the midcycle LH/FSH surge. In contrast, the FSH could stimulate granulosa cell progesterone secretion if the cells were harvested both prior to (60- to 100-fold stimulation) and during the midcycle LH/FSH surge (10- to 60-fold stimulation; P less than 0.05). It can be concluded that exposure of the preovulatory monkey follicle to the midcycle LH/FSH surge alters its responsiveness to FSH in terms of estrogen secretion.     

Biologic action of promestriene on the genital tract of the castrated rat

The study assessed the activity of promestrien administered by a local vaginal route in 16 rats castrated 60 days previously. The dose used was 5 mg per day for 10 days. Following treatment, animals were sacrificed and histological analyses revealed a remarkable proliferation of the vaginal epithelium, inhibition and restored trophism of the strome, whereas no estrogen stimulus was observed at the level of uterine mucous. This confirms the exclusively local action of promestrien which does not provoke undesirable side effects on other segments of the genital tract, and allows the therapeutic use of the hormone in different gynaecological pathologies.     

Elevation of plasma immunoreactive luteinizing hormone releasing hormone in hyperprolactinemic-amenorrheic women on bromocriptine therapy

There is evidence to suggest that abnormalities in the secretion of prolactin (PRL) in patients with the hyperprolactinemia-amenorrhea syndrome are due to hypothalamic dysfunction. In an attempt to further define the inhibitory effect of excessive PRL release on luteinizing hormone releasing hormone (LHRH) and luteinizing hormone (LH) secretory patterns in human plasma, four amenorrheic women with known hyperprolactinemia were studied before and during bromocriptine (BRCR) therapy. Ten-minute blood samples collected with a continuous withdrawal pump for two hours were analyzed for immunoreactive LHRH (IR-LHRH), LH and PRL using previously established radioimmunoassay procedures. Three patients showed a significant rise in mean IR-LHRH plasma levels coincident with a significant decrease in mean PRL concentrations five days to two weeks following BRCR therapy, whereas mean LH titers increased significantly in only one patient. One patient showed no increase in IR-LHRH or LH with BRCR therapy and failed to show a decrease in serum PRL to normal levels after five days of this treatment. A defect in the control of PRL release in these patients seemed to result from the inability of dopaminergic inhibition to be mediated effectively and seemed to be associated with altered secretion of LHRH.     

Effect of D-leucine-6-luteinizing hormone-releasing hormone ethylamide in patients with hypogonadotropic hypogonadism with anosmia.

Four men with hypogonadotropic hypogonadism and anosmia were tested with acute intravenous injections of luteinizing hormone-releasing hormone (LH-RH) and D-leucine-6-LH-RH-ethylamide (D-L eu-6-LH-RH-EA) with a 1-week interval. Each patient was then treated with this drug for 60 days and tested again after this period with an intravenous injection of D-L eu-6-LH-RH-EA. The administration of LH-RH resulted in a significant increase in the LH level in only one patient and in follicle-stimulating hormone (FSH) and testosterone increases in none. The analog D-Leu-6-LH-RH-EA resulted in significant increases in LH levels in two patients, in FSH levels in three, and in testosterone levels in one. Results obtained after treatment were closely similar to those observed before treatment. Clinical improvement in terms of increased libido, erection, pubic hair growth, and testicular size was observed. D-Leu-6-LH-RH-EA could be useful in the treatment of patients with hypogonadotropic hypogonadism, a possibility deserving further studies.     

Quantitative analysis of cellular proliferation and differentiation of the human seminiferous epithelium in vitro

The aim of the present work was to quantitate the temporal and stage-specific effects of follicle-stimulating hormone (FSH) and testosterone on the proliferation and differentiation capacities of the human seminiferous epithelium. Seminiferous tubule fragments were kept in culture for 28 days and 5-bromo-2'-deoxyuridine incorporation was used to determine cell proliferation. Data demonstrated a gradual loss of germ cells during the culture period, no decrease in Sertoli cell numbers, and maintenance of the general architecture of the seminiferous tubules. Both FSH and testosterone increased germ cell survival, spermatogonia proliferation, and germ cell differentiation, especially during the first week of culture. At the end of the first week, differentiation of spermatocytes was observed, especially when 50 IU/L FSH and 1 μmol/L testosterone were used. In conclusion, using this methodology, it was possible to quantify germ cell proliferation and differentiation, in a reproducible way, with results compatible with the timing of human spermatogenesis in vivo.     

Release rate of testosterone and estrogens from polydimethylsiloxane implants for extended periods in vivo compared with loss in vitro.

Release rates of testosterone, estrone, and estradiol placed in chambers made from polydimethylsiloxane (PDS) tubing (Dow Corning "Silastic," 3.35 mm ID x 4.65 mm OD) were studied in 14 freemartin cattle with minimal or non-detectable endogenous hormone secretion, and in 0.9% saline:methanol (1:1) baths shaken at 38 degree C. Eighty-seven implants, varying in length from 2 to 10 cm, were placed in 14 animals for 27 to 235 days. The average release rates +/- standard errors, in microgram/cm/day, were testosterone, 55.9 +/- 2.4, estrone, 12.6 +/- 1.8, and estradiol, 11.1 +/- 1.1. A relatively constant release rate was found over the period of time studied and sufficient steroid remained for potential release over periods exceeding 1 year. The dose of hormone delivered was sufficient to increase mounting activity in testosterone-treated animals and estrual activity in those receiving estrogens. Corresponding release rates in vitro for four 10-cm implants containing either testosterone, estrone, or estradiol were 94.3 +/- 1.9, 15.5 +/- 0.7, and 12.7 +/- 0.6 microgram/cm/day, respectively. The general magnitude of release rate in animals could be predicted from laboratory tests.