Biological manifestations of the andropause

Aging in men is accompanied by signs of decreased virility and fertility: testicular volume, muscle mass, pilosity, sexual activity, daily production of spermatozoa, plasma testosterone levels, the number of Leydig cells and blood supply of the testicles decrease significantly with age. The pulsatility of LH levels in young and elderly men has been studied by taking plasma samples with an interval of 10 minutes for 12 hours. Age alterations at the hypothalamic-pituitary levels of testosterone production and an increase in gonadostat sensitivity to hormonal influences realized via the feedback mechanism are found. Whenever androgen therapy of eugonadal elderly men is considered, a sufficient dosage should be administered, as otherwise, due to the feedback effects, administered androgens, suppressing endogenous secretion might not increase plasma-testosterone levels. Changes in neurotransmitters and neuromodulators play a determinant role in the sexual behaviour of elderly men.     

Age-related changes of plasma steroids in normal adult males

Plasma cortisol, 17-hydroxyprogesterone (17-OH-P), testosterone (T), 5α-dihydrotestosterone (DHT, estrone (E₁) and estradiol (E₂), were measured in 94 normal adult men aged between 20–99. using RIA methods after chromatographic separation of steroids on Sephadex LH-20 columns.All plasma steroids except 17-OH-P, were age dependent: cortisol, testosterone and DHT decreased significantly with age, whereas estrone and estradiol were significantly increased in elderly men. Cortisol, testosterone. T/DHT ratio and estradiol levels were significantly correlated with age.The age related changes of plasma steroids in elderly men, were suggestive of decreased cortisol secretion, and decreased testicular function with increased peripheral conversion of androgens into estrogens. Testosterone was positively correlated with its precursor (17-OH-P) and respectively its peripheral metabolites (DHT and E₂). The negative correlation between estrone and 17-OH-P found in elderly men, suggested that increased estrogen level in aging males may be considered able to inhibit the testicular androgen production.     

Reduced calcitonin reserve in young hypogonadic osteoporotic men

Calcitonin is a potent inhibitor of bone resorption and in both sexes, plasma levels progressively decrease with age: therefore, a relative deficiency of calcitonin may be involved in the pathogenesis of osteoporosis in the elderly. Calcitonin plasma levels of young hypogonadic men with osteoporosis are significantly lower than controls: the hypothesis that the decreased calcitonin plasma levels in the elderly are due to a reduced secretory capacity of the "C" cells of the thyroid gland, related to age, does not explain the low calcitonin plasma levels found in young hypogonadic osteoporotic men. Our hypothesis is that gonadal steroid deficiency may participate in the mechanisms regulating calcitonin secretion. Therefore, we studied ten males affected by hypogonadotropic hypogonadism and ten normal men, of comparable age, as controls: we measured plasma levels of testosterone, 17 beta estradiol, androstenedione and calcitonin, and the response of calcitonin to an i.v. bolus of pentagastrin, a well known "C" cells stimulatory drug. Testosterone and calcitonin plasma levels and the response of calcitonin to pentagastrin were also evaluated after 6 months of replacement therapy with testosterone. Basal levels of testosterone, 17 beta estradiol, androstenedione and calcitonin, and the response of calcitonin to pentagastrin, are significantly lower in our patients than in controls, demonstrating that hypogonadotropic hypogonadic subjects have a lower secretory reserve of calcitonin. After testosterone therapy the basal calcitonin plasma levels and its response to pentagastrin stimulus did not differ from controls, suggesting that gonadal steroids influence the calcitonin secretion and reserve. Our data cannot clarify whether osteoporosis of hypogonadotropic hypogonadic patients is related to androgen or estrogen deficiency; however, they suggest that the mechanisms by which gonadal steroid influence bone metabolism may involve calcitonin secretion.     

Effects of exogenous testosterone on testicular luteinizing hormone and follicle-stimulating hormone receptors during aging

During aging, the male Japanese quail exhibits a loss of fertility, increased morphological abnormalities in the testes, and a higher incidence of Sertoli cell tumors. Although there is a coincident loss of reproductive behavior, plasma androgen levels remain high until testicular regression occurs in association with senescence. The purpose of this study was to compare mean specific binding of chicken luteinizing hormone (LH) and follicle-stimulating hormone (FSH) as a measure of testicular receptors during identified stages during aging. Males were categorized according to age (young = 9 months, middle aged = 24 months, or old = 36+ months) and sexual behavior (active or inactive). Testicular samples were collected immediately after perfusion with 4% paraformaldehyde from the following groups: young active (n = 8), young photoregressed (n = 5), young photoregressed plus testosterone implant (n = 4), middle-aged active (n = 8), middle-aged inactive (n = 4), old inactive (n = 5), and old inactive plus testosterone implant (n = 6). A crude plasma membrane fraction was prepared from the testes of each bird and an aliquot deriving from 10 mg of testicular tissue was used for binding assay. Specific binding of labeled LH or FSH was expressed as percentage of total radioactive hormone. Results showed significant (P < 0.05) age-related decreases in both FSH and LH receptor numbers. The highest FSH binding was found in young and middle-aged active males, with low binding in old inactive males. Testicular LH binding decreased during aging, with a sharp decrease in middle-aged males, which was similar to old males. Testosterone implants weakly stimulated FSH and LH binding in old males. Both LH and FSH binding decreased in photoregressed young males. However, testosterone implants stimulated increased LH binding, but did not affect FSH binding in young photoregressed males. These results provide evidence for separate regulation of testicular LH and FSH receptors, with testosterone stimulation of LH receptor, but not FSH receptor number in young males. However, during aging there appears to be a loss of this response, potentially because of the reduced efficacy of testosterone stimulation, thereby implying a diminished capacity for response with aging.     

醋酸棉酚对大鼠睾丸HCG反应性及其受体功能的影响

给大鼠灌服醋酸棉酚30 mg/kg/d,每周6次,持续4周。给药2周后,血浆睾酮水平显著下降并持续至第4周,同时间质细胞呈萎缩性改变。醋酸棉酚明显抑制成年大鼠对HCG反应性,使睾丸LH/HCG受体亲和力下降,受体数目略有减少。结果提示,醋酸棉酚抑制大鼠睾丸酮的产生及降低成年大鼠睾丸对HCG的反应性,推测其机制是由于醋酸棉酚干扰了睾丸HCG受体功能而造成的。     

Studies on the prostate glands of adult inbred LSH hamsters.

We have studied several parameters of prostate function in two inbred lines (2.4 and SsLak) of young and old LSH hamsters. These included weight, acid phosphatase, and [3H]testosterone uptake as influenced by age, castration, and androgen treatment. In the hamster, prostatic acid phosphatase concentration was found to vary inversely with androgen levels, contrary to the usual assumption that this enzyme is androgen dependent. Prostatic uptake of tritiated testosterone was enhanced by castration and by treatment of castrates with doses of androgen which induced a moderate increase in gland size. With advancing age, the prostates of LSH hamsters (both strains) became atrophic rather than hyperplastic, in contrast with a previous report (1). This atrophy appeared to be a consequence of decreased testicular function. The LSH hamsters appear to be a suitable model for the study of senescent changes in the male reproductive system.     

The effect of human chorionic gonadotropin on plasma steroid levels in young and old men

Seven men, 21–30 years old, and six men, 72–90 years old, were given human chorionic gonadotropin (HCG), 3,000 units a day for four days. The concentration of 17β-estradiol (1), estrone and testosterone were measured in plasma samples drawn before and during the course of HCG administration. The administration of HCG resulted in higher levels of both 17β-estradiol and testosterone in the younger as compared to the older men although the percentage increases over baseline levels were similar in both groups. HCG administration resulted in similar, absolute and relative increases of estrone in both young and old men. The levels of 17β-estradiol were higher on day 3 as compared to day 5 in young men. The relative ability to respond to exogenous gonadotropins appears to be preserved despite ageing and loss of libido and potentia. The absolute response is, however, somewhat less in old men as compared to young.     

Role of testosterone levels and of hypophysis in the HCG-induced modifications of the 7 alpha-hydroxylation and 5 alpha-reduction processes in incubated rat testes

The role of a direct or a hypophysis-mediated influence of increased testosterone levels in the effects of a long-term high-dose HCG administration (10 IU/day) upon the 7 alpha-hydroxylation and 5 alpha-reduction activities of incubated testes of mature rats was investigated. Administration of high doses of HCG to hypophysectomized rats resulted in the same metabolic changes as in normal rats, namely, a large decrease in the 7 alpha-hydroxylation and an increase in the 5 alpha-reduction processes. Administration of testosterone-propionate (0.2 mg and 20 mg/day) for several days to hypophysectomized rats and to normal rats receiving and substitutive dose of 1 IU HCG/day, did not modify the testicular metabolization pattern. These findings indicate that the decrease in testicular 7 alpha-hydroxylation activity induced by long-term administration of high doses of HCG is probably not mediated by the hypophysis nor by the extracellular testosterone levels.     

Corticosteroids affect the testicular androgen production in male common carp (Cyprinus carpio L.).

Our previous experiments to study the effect of stress adaptation on pubertal development in carp showed that repeated temperature stress and prolonged feeding with cortisol-containing food pellets, which mimics the endocrine stress effects, retarded the first waves of spermatogenesis and decreased 11-ketotestosterone (11KT) plasma levels. The objective of the present study was to investigate whether the decrease in plasma 11KT is caused by a direct effect of cortisol on the steroid-producing capacity of the testis or by an indirect effect, such as a decrease in plasma LH. Pubertal and adolescent isogenic male common carp (Cyprinus carpio L.) were fed with either cortisol-containing food pellets or control food pellets over a prolonged period. Our results indicate that cortisol has a direct inhibitory effect on the testicular androgen secretion independent of the LH secretion. Furthermore, the pubertal period is critical to the influence of cortisol regarding testicular androgen secretion, because the effect is no longer observed at adolescence.     

Radioimmunoassay of 17β-hydroxy-5α-androstan-3-one, 4-androstene-3,17-dione,dehydroepiandrosterone, 17-hydroxyprogesterone and progesterone and its application to human male plasma

RIA methods for the measurement of 5α-dihydrotestosterone (DHT), androstenedione (Δ4), dehydro-epiandrosterone (DHEA), 17-hydroxyprogesterone (17-OHP) and progesterone (P) have been developed and applied to the study of factors determining the levels of these steroids in males.It has been shown that DHT, 17-OHP and DHEA levels are lower in elderly males than in young (<50 years) subjects, Δ4 and P levels remaining constant. All steroids, with the exception of DHT, show significant nyctohemeral variations: upon ACTH stimulation T levels decrease, whereas Δ4 and DHEA, 17-OHP and P increase significantly. Dexamethasone has the reverse effect, although T levels do not decrease. HCG stimulation results in a significant increase of T, DHT, 17-OHP and Δ4 levels, whereas DHEA and P levels are hardly influenced. In a small group of orchidectomized males, all steroids studied, with the exception of P were significantly lower than in normal males of similar age.It is concluded that in males T, DHT and 17-OHP have an almost exclusive testicular origin; Δ4 has a mixed testicular and adrenal origin, whereas DHEA has a predominant and P an exclusive adrenal origin in males.     

The Immediate Effect of HCG Upon Plasma Testosterone Levels in the Bull

No effect of HCG upon the plasma testosterone level in 16 bulls was seen the first 5 or 15 min after injection. In 8 bulls receiving 750 or 1500 i.u. HCG by intravenous injection a lag time of 30 min occurred before plasma testosterone response could be measured. The high plateau level of plasma testosterone concentration was reached approximately 1 h after injection. Following intramuscular injection of 750 i.u. or 1500 i.u. HCG a lag time of 45–60 min was observed for the testicular response measured as elevated plasma testosterone level. The high plateau levels of plasma testosterone in these bulls were reached approximately 1½ h post injection.     

L'andropause

In men, aging is associated with progressive impairment of testicular function. Decrease in total testosterone levels with aging has been reported in many studies in normal healthy men. This decrease has a primarily testicular origin, as shown by decreased number of Leydig cells in histological studies, increased basal gonadotropin levels and decreased response to hCG. A greater decrease in bioavailable testosterone rather in than total testosterone concentrations is explained by the age-dependent increase in the SHBG concentration. Although immunoreactive gonadotropin levels are higher than in young men, a relative alteration of bioactive gonadotropin secretion by the pituitatry occurs in ederly men. Althought the definitive demonstration of an alteration of GnRH secretion by the hypothalamus cannot be established in healthy ederly men, such an alteration might be responsible for a decompensation of the testicular function in case of intercurrent illness. Increased FSH and decreased inhibin plasma levels are indicating a similar alteration in seminiferous tubules as directly demonstrated by histological data showing a decrease of Sertoli cell number and daily sperm production with aging. Although the incidence of sexual dysfunction increases with aging, the relationship between sexual behaviour and testicular endocrine function remained a mater of controversy. A threshold of testosterone action on sexual behavior might be responsible for the difficulty in establishing this relationship. Although some controled studies are available, the risk-to-benefit balance of androgen substitution in older male remained a controversial issue. A positive effect on sense of well-being and/or libido has been and the lack of adverse effect on lipid and carbohydrate metabolism had been demonstrated in short term studies, however, the role of androgens in the benign hypertrophy of the prostate and in stimulating the growth of latent prostate adenocarcinoma remained to be more clearly established by longterm controlled studies.     

Gonadotropin receptors in rat during altered testicular function

Specific testicular binding of 125I-HCG and plasma levels of testosterone are decreased in rats three weeks after hypophysectomy or adrenalectomy plus thyroidectomy but they are not changed after adrenalectomy or thyroidectomy. The affinity of gonadotropin receptors to HCG is not altered.     

Plasma androgens and their association with the reproductive cycle of the male fence lizard, Sceloporus undulatus

Seasonal variation in plasma androgen levels was determined for males of the lizard Sceloporus undulatus. Plasma androgens peaked in both spring and in fall prior to brumation, coinciding with periods of maximum testicular enlargement and advanced spermatogenic development. Plasma androgen concentrations were minimal during summer, following testicular regression and breeding in spring. Epididymal masses were positively correlated to plasma androgen levels, while abdominal fat body masses demonstrated a negative correlation.     

Simultaneous measurements of testosterone and three 5a -reduced androgens in the venous effluent of immature rat testes in situ

Simultaneous measurements were made by radioimmunoassay of testosterone, 17β-hydroxy-5 a -androstan-3-one (DHT), 5 a -androstane-3 a, 17 β -diol (3a-diol), and 5 a-androstane-3 β, 17β-diol (3β-diol) in the testicular venous plasma (TVP) and peripheral plasma (PP) of 30, 45, and 55 day old rats. At 30 days of age, the preponderant androgen in both plasmas was 3a-diol but testosterone predominated by day 55. Testosterone levels increased with age in both TVP (6.39, 15.08, and 54.93 ng/ml on days 30, 45, and 55 respectively) and PP (0.13, 0.56, and 1.02 ng/ml on days 30, 45 and 55 respectively) whereas 3a-diol concentrations decreased in TVP (48.07 ng/ml, day 30; 24.85 ng/ml, day 55) though not peripherally (range: 0.41–0.52 ng/ml). DHT was low in both TVP and PP and appeared to rise only slightly although the increase was not statistically significant. Levels of 3β-diol remained low and unchanged. These observations suggest that the total androgen content of the venous effluent from the prepubertal rat test is is quite high and that significant changes in peripheral interconversions of androgens are occurring during sexual maturation.     

Neuroendocrine regulation of pulsatile luteinizing hormone secretion in elderly men

Leydig cell function is driven by LH, secreted in a pulsatile manner by the anterior pituitary in response to episodic discharge of hypothalamic LHRH into the pituitary portal circulation, under control of a yet to be defined neural mechanism, the "hypothalamic LHRH pulse generator". The normal aging process in elderly men is accompanied by a decline in Leydig cell function. Whereas primary testicular factors undoubtedly play an important role in the decrease of circulating (free) testosterone levels with age, recent studies demonstrated that aging also affects the central compartment of the neuroendocrine cascade. Hypothalamic alterations comprise changes in the regulation of the frequency of the LHRH pulse generator with an inappropriately low frequency relative to the prevailing androgen impregnation and opioid tone, and with an increased sensitivity to retardation of the LHRH pulse generator by androgens. As observed by some authors in basal conditions and by others after endocrine manipulations. LH pulse amplitude seems also to be reduced in elderly men as compared to young subjects. This is most probably the consequence of a reduction in the amount of LHRH released by the hypothalamus. Indeed, challenge of the gonadotropes with low, close to physiological doses of LHRH in young and elderly men reveals no alterations in pituitary responsiveness when looking at either the response for immunoreactive LH or bioactive LH. Deconvolution analysis on data obtained after low-dose LHRH suggests a markedly prolonged plasma half-life of LH in elderly men, a finding which may explain the paradoxical increase of mean LH levels in face of the reduced or unchanged frequency and amplitude of LH pulses.     

Possibility of adrenal-testicular interaction as indicated by plasma androgens in response to HCG in men with normal, suppressed and impaired adrenal function.

In order to assess possible adrenal-testicular interactions in vivo, adrenal and testicular plasma androgens (testosterone, delta4-androstenedione, dehydroepiandrosterone and dehydroepiandrosterone sulfate) were measured by specific radioimmunoassays before and after stimulation with HCG in men with normal, dexamethasone suppressed and impaired adrenal function. It was found that men with Addison's disease, in whom circulating dehydroepiandrosterone sulfate levels amounted to 1/10 of normal values, had a decreased response of testosterone to HCG. Simultaneously, the Addison patients and the men under dexamethasone had only an increase of delta4-androstenedione but not of dehydroepiandrosterone, while normal men showed an almost equal increase in both precursors under HCG. The results are interpreted as demonstrating that the delta5-pathway in the testis becomes less important under adrenal suppression and in Addison's disease due to a lack of substrate (possibly dehydroepiandrosterone sulfate) from the adrenals.     

The effect of prenatal treatment with busulfan on in vitro androgen production by testes from rats of various ages

Treatment of rats with busulfan in utero severely depletes the germ cell population of the seminiferous tubules. These studies have examined the in vitro capacity of testicular tissue and Leydig cells from such testes to secrete androgens. Leydig cells were identified by staining for 3 beta-hydroxy steroid dehydrogenase. Rats were studied at several ages to identify any developmental changes in the androgen-secreting capacity of control and treated gonads. At 30 days of age, no effect of treatment on serum androgen was found. At 60 and 90 days of age, treatment caused decreased androgen and increased LH content of the serum. At 12, 30, 60, and 90 days of age, the amount of androgen secreted per milligram of testicular tissue in response to LH was higher in busulfan-treated rats. Leydig cells from 60- and 90-day-old rats which had received busulfan were also hyperresponsive to LH. It was concluded that Leydig cells from testes essentially devoid of germ cells were hyperresponsive to LH. Serum androgen levels were decreased yet androgen production per Leydig cell was increased. A possible explanation of this apparent paradox is that busulfan treatment resulted in decreased numbers of Leydig cells in the gonads.     

Androgen and gonadotropin effects on male mate calls in South African clawed frogs, Xenopus laevis

Mate calling is a prominent reproductive behavior of male South African clawed frogs. Calls consist of alternating slow- and fast-amplitude-modulated trills. Each trill is made up of a series of clicks. The effects of administration of exogenous gonadotropin and androgen on mate calling were studied in male Xenopus laevis. Males were paired with unreceptive female frogs to elicit maximal calling. The amount of time each animal spent calling during the testing period, the peak fundamental frequency of the calls, the rate of calling, and the interclick interval (ICI, a measure of the temporal patterning of the calls) were measured in intact, castrated, and hormone-replaced frogs. Injection of human chorionic gonadotropin (HCG) into intact frogs increased the amount of time spent calling and the ICI relative to measures taken after water injection. Castrated males did not call even when given HCG. Testosterone and dihydrotestosterone treatment reinstated calling in castrates and increased circulating levels of androgens. When androgen-replaced castrated males were injected with HCG, the amount of time spent calling increased and approached levels of intact, HCG-injected males. The above results suggest that androgens are necessary for the production of calls. Gonadotropins appear to play an important role in mate calling, a role at least partly independent of effects on testicular androgen synthesis.     

Effects of ovariectomy of the dogfish Scyliorhinus canicula L. on circulating levels of androgen and oestradiol and on pituitary gonadotrophin content

Levels of androgens and oestradiol in dogfish, Scyliorhinus canicula , plasma, measured by radioimmunoassay, decrease markedly after ovariectomy (OVX). The magnitude and latency of these responses varies according to length of captivity preceding the operation and the time of year at which ovariectomy is performed. Androgens are cleared more rapidly than oestradiol from plasma following OVX, and levels of both steroids are reduced after laparotomy (LAP) in November but not in March. An extract of ventral lobes of the dogfish pituitary induces a significant increase in the circulating androgen levels in LAP fish but not in OVX fish, indicating that the ovary is the major site of androgen production in response to stimulation by dogfish gonadotrophin. Intramuscular injections of oestradiol benzoate (2.5 mg per fish) into LAP fish produce a significant reduction in the gonadotrophin content of the pituitary ventral lobe, measured in the quail testicular cell bioassay, when compared to LAP and unoperated controls. However, no significant changes in the gonadotrophin content of the ventral lobe were seen five days after OVX in June.