Altered gonadal hormone level and constant light-induced stress interfere with the response of the adrenal medulla to oxytocin.

The effect of oxytocin (0.25 IU/100 g per day) on the adrenal medulla was examined in intact, intact estrogen-treated, castrated and castrated testosterone-treated adult male Wistar rats. Stereological analysis of the gland (N = 5 rats per group) revealed that in intact animals the number of chromaffin cells (x10(3)) was significantly increased after 3-day (saline: 467.6 +/- 27.4; oxytocin: 567.6 +/- 28.9) or 7-day (saline: 486.2 +/- 39.1; oxytocin: 618.7 +/- 36.8) oxytocin administration. During 7 days of recovery after the 7-day treatment, the chromaffin cell number returned to the control level (saline: 491.4 +/- 12.6; oxytocin: 554.4 +/- 28.7). The effect of oxytocin on chromaffin cell number was also observed in rats simultaneously injected with estradiol (0.3 micrograms/100 g per day) for 10 days (estradiol: 454.3 +/- 32.8; estradiol+oxytocin: 576.1 +/- 25.0), as well as in 10-day castrated rats (saline: 594.7 +/- 22.7; oxytocin: 765.3 +/- 33.1). Testosterone replacement (0.6 mg/100 g per day) abolished the medullary response to oxytocin (testosterone+saline: 528.5 +/- 24.7; testosterone+oxytocin: 620.8 +/- 56.0). There was a 20% rise in adrenal dopamine content (from 0.236 +/- 0.015 to 0.283 +/- 0.015 microgram per pair of glands; N = 9-12) in intact rats injected with oxytocin for 3 days. Oxytocin had no effect on any of the catecholamine levels in adrenal glands of rats exposed to stress induced by constant lighting. The present data indicate that the proliferative response of chromaffin tissue to oxytocin depends on the gonadal hormone level and the basal activity of the adrenal medulla.

Opposite effects of dexamethasone and ACTH on the adrenal cortex response to ethane dimethanesulphonate (EDS).

Recently we have reported that ethane dimethane-sulphonate (EDS), the Leydig cell cytotoxin, caused marked atrophy of the adrenal cortex of adult male rats. The aim of this work was to examine whether a 9-day treatment with dexamethasone (0.25 mg/kg/d) or ACTH (40 IU/kg/d), which started 4 days prior to administration of a single dose of EDS (75 mg/kg), influenced the response of the inner adrenocortical zones to the toxin. On day 15 after administration of EDS, adrenal weight was significantly decreased in saline treated rats, but glandular and serum corticosterone levels were not altered. In dexamethasone-suppressed rats, the effect of EDS was augmented; an additional decrease in adrenal weight was accompanied by reduced adrenal and serum corticosterone levels. In ACTH-treated animals EDS was ineffective. These results demonstrate that the deleterious effects of EDS on rat adrenal cortex can be prevented by ACTH and potentiated by dexamethasone.

Treatment of pregnant females with dexamethasone influences postnatal development of the adrenal medulla.

In the light of the mutual dependence between the adrenal cortex and medulla, the aim of this work was to examine whether glucocorticoid treatment of pregnant rats affects the development of the adrenal medulla of their offspring in the postnatal period. Pregnant rats were treated with dexamethasone (Dx) in a daily dose of 0.3 mg Dx/kg b.w. during days 16-20 of gestation. The structure and function of the adrenal medulla of their 14-day-old offspring were estimated on the basis of the morphometric parameters of the gland, chromaffin cell mitotic index and adrenal gland adrenaline content. Stereological analysis was carried out at the light microscopic level, the mitotic index was determined by counting the number of metaphase arrested chromaffin cells following the administration of vincristine-sulphate, whereas adrenaline content in the adrenal gland was measured fluorimetrically. Plasma ACTH concentrations of the offspring were also determined by RIA. Long term Dx treatment of pregnant rats caused a significant decrease of the total volume of adrenal chromaffin tissue in the 14-day-old offspring as well as a reduction in the number of chromaffin cells and the average cell and nuclear volumes. The proliferative activity of the chromaffin cells was also lower than in the control offspring. These changes were accompanied by a significantly reduced adrenaline content in the adrenals. The results of this work show that glucocorticoid excess during the period of pregnancy when the fetal adrenal medulla is formed has a strong inhibitory effect on the adrenal medulla of the offspring at the age of 14 days.

beta-Adrenergic receptor manipulation and acid phosphatase and zinc levels in the ventral prostate of the adult rat.

The influence of 15-day treatments with the beta-adrenergic receptor agonist isoproterenol (120 micrograms/kg/d) or the antagonist propranolol (1.00 mg/kg/d) on acid phosphatase and zinc levels in the ventral prostate was examined in intact rats, rats simultaneously injected with dexamethasone (0.25 mg/kg/d) and animals chemically castrated with a single dose of ethane dimethanesulphonate (75 mg/kg). Isoproterenol-treatment significantly increased acid phosphatase concentration in the ventral prostate of intact rats, whereas propranolol prevented a glandular zinc loss induced by dexamethasone administration. These results demonstrate that the levels of both biochemical parameters in the prostate can be altered by beta-adrenergic receptor manipulation. The responsiveness of the two secretory processes is different and depends on the functional status of the ventral prostate.

Propranolol treatment affects ventral prostate blood vessels and serum testosterone concentrations in adult rats.

The influence of prolonged beta-adrenergic receptor blockade on stereologic parameters of the ventral prostate and serum testosterone concentrations was examined in adult rats injected with propranolol (0.1 or 0.4 mg kg-1/day) for 15 or 30 consecutive days. Both doses of propranolol reduced the relative and absolute volume of the ventral prostate blood vessels. This effect was prevented by simultaneous administration of urapidil, an alpha 1-adrenergic receptor antagonist, indicating that a compensatory vasoconstriction took place as a consequence of propranolol treatment. Serum testosterone concentration was significantly increased following the 30-day application of the lower dose of the drug. These results show that prolonged administration of propranolol, although not affecting the epithelial component of the gland, may indirectly influence prostatic function by reducing the blood flow to the gland.

Differential effects of chronic propranolol treatment on the phenotypic profile of thymocytes from immature and adult rats.

To elucidate a putative role of beta-adrenoceptors in the modulation of intrathymic T-cell maturation, the expression of major differentiational antigens (CD4/CD8 and TCR alphabeta) on the thymocytes from both immature (aged 21 day at the beginning of the treatment) and adult (aged 75 days at the beginning of treatment) male rats subjected to a 15-day-long propranolol treatment (0.40 mg/100 g/day, s.c.) was analyzed by two- and one-color flow cytometry, respectively. Rats of matched age injected with saline served as controls. The propranolol treatment in immature but not adult rats caused a significant reduction in both the relative thymus weight and total thymocyte yield. In addition, a significant increase in the percentage of CD4+ 8+ double-positive cells, with a proportional decrease in the relative proportion of CD4+ 8- single positive cells, was found in immature rats. In contrast, a slight but significant decrease in the percentage of CD4+ 8+ cells with a parallel increase in the relative proportion of CD4+ 8- cells was found in adult rats. In both groups of rats, the percentage of TCR alphabeta(total) thymocytes was increased: in immature rats this was due to an increase in the percentage of TCR alphabeta(low) thymocytes, while in the adult rats it reflected a rise in the relative proportion of TCR alphabeta(high) cells. In conclusion, the study revealed that propranolol treatment in both immature and adult rats alters the relative proportion of CD4+ 8+ and CD4+ 8- thymocytes, but in opposite fashion, and the data suggest that this treatment affects distinct fractions within the population of CD4+ 8+ thymocytes with respect to expression of TCR alphabeta. The results also indicate that, regardless of rat sexual maturity, the development of thymocytes towards CD4- 8+ T-cells is relatively insensitive to long-lasting beta-adrenoceptor blockade.

Rat accessory sex glands response to oxytocin under different light regimens.

Effects of oxytocin (0.25 IU OT/100 g/d for 3 days) on accessory sex glands structure and catecholamine content were examined in rats kept on two different light regimens. In 12 h light/12 h dark conditions ventral prostate responded to OT by a regression of the epithelial component and an increase in dopamine content. Coagulating gland structure was not affected, but noradrenaline content of the seminal vesicle+coagulating gland complex was enhanced. Constant lighting per se caused atrophic changes in the prostatic epithelium, evident by decreased total volume and by increased percentage of acini containing cuboidal epithelium. OT treatment considerably prevented the epithelial atrophy without affecting catecholamines level. In the seminal vesicle+coagulating gland complex it reduced the dopamine content. Since this light regimen elevated the plasma ACTH, the altered accessory sex gland response to OT seems to be due to the stress-induced changes in the neuro-endocrine factors conditioning their function. The effects of OT on accessory glands catecholamine content indicate interferences with their autonomic control.

Mitotic activity and cell deletion in ventral prostate epithelium of intact and castrated oxytocin-treated rats.

Previous study has shown that oxytocin (OT) attenuated the postcastration regression of ventral prostate epithelium in rats. To decide if this effect is associated with stimulation of cell division or improvement of cell survival, metaphase index of secretory cells and frequency of apoptotic bodies in the epithelium, combined with stereological parameters, were determined in the present study. OT was injected subcutaneously in a dose of 0.25 IU/100 g/d during 5 postorchiectomal days and the same treatment was applied to intact rats. Only the prostate of castrated animals responded to OT. They had greater total volume of the epithelium, total number and metaphase index of secretory cells, but lower frequency of apoptotic bodies. These findings demonstrate the ability of OT to stimulate mitotic activity and to diminish mortality of secretory cells caused by orchiectomy.

The effect of dexamethasone on the adrenal gland in fetal and neonatal rats.

The development and regeneration of the adrenal glands were examined by stereological and morphological methods in 20-day-old fetal, as well as 3-day- and 14-day-old neonatal male rats born to dams treated with dexamethasone (Dx) on day 16 of gestation. In the fetuses and 3-day-old rats, zona fasciculata (ZF) and zona reticularis (ZR) were analyzed as one inner zone (IZ), while in 14-day-old animals they were analyzed separately. Single Dx treatment (1.5 mg/kg b.w.) of the dams led to atrophic changes in the adrenal cortex of the fetuses. These changes were visible to a certain degree up to the 14th neonatal day. Administration of Dx to pregnant rats induced a significant decrease of both adrenal weight and volume, as well as the volume of zona glomerulosa + capsule (ZG + C) and IZ, both in fetuses and 3-day-old rats. This was due to a decrease in the number but not the volume of cortical cells. Also, necrotic cortical cells, infiltrations and resorption zones accompanied by the presence of macrophages, giant cells and lymphocytes were observed. In 14-day-old animals, the degree of atrophic changes in the adrenal cortex was reduced. Changes were observed only in ZR which was decreased in volume resulting from both a significant decrease of the volume and number of cortical cells. Then number of macrophages was somewhat increased, while giant cells were not present. However, the total number of parenchyma cells in ZG was increased, pointing to the possibility of renewal of cortical cells within this zone. The results of the present study demonstrate that even a single Dx dose given to pregnant rat during the period critical for the development of the hypothalamo-pituitary-adrenal system in the fetuses leads to marked changes in the structure and function of the fetal adrenal glands which are partially maintained up to the 14th day of postnatal life.

The influence of prolonged dexamethasone treatment of pregnant rats on the perinatal development of the adrenal gland of their offspring.

The effects of prolonged dexamethasone (Dx) administration to pregnant rats on the structure and function of the adrenal glands of fetal and neonatal offspring have been investigated by combined stereological and ultrastructural methods, as well as by metaphase index determination. Pregnant rats were injected subcutaneously with Dx (0.3 mg/kg body weight/day) during 5 days, starting from day 16 of gestation. The dams and their fetuses were killed 24 hr after the last injection. The neonatal offspring were killed in the same way on the 3rd and 14th day of life. Because in fetal and 3-day-old neonatal rats zona reticularis (ZR) was poorly defined and could not be clearly seen as a separate zone, zona fasciculata (ZF) and ZR were analyzed as one, inner zone (IZ). In 14-day-old rats ZF and ZR were analyzed separately. Proliferative activity of adrenocortical cells was estimated following the application of Vincristine sulphate. Dx treatment of pregnant rats induced a marked decrease of fetal adrenal gland volume and the volumes of zona glomerulosa + capsula (ZG + C) and IZ as the consequence of atrophic changes in the gland and reduction of the average volume and total number of adrenocortical cells. Similar morphometric changes were found in 3- and 14-day-old pups. However, in 3-day-old animals the number of cortical cells in the ZG was increased, whereas on the 14th postnatal day cortical cell number remained decreased only in the ZF. The multinuclear giant cells, numerous lymphocytes, and the resorption zones, present in the adrenal cortex of fetuses and 3-day-old pups of both experimental and control dams, were not seen in 14-day-old offspring. These results demonstrate that prolonged treatment of pregnant rats with Dx in the period when intensive differentiation of the fetal hypothalamo-hypophyseal system takes place inhibits proliferative activity of adrenocortical cells and evokes considerable atrophic changes in the adrenal glands of offspring from 20 days gestation to 14 days after birth. The histological appearance of the adrenal cortex and the ultrastructure of adrenocortical cells suggest that cortical cell function was inhibited.

Ventral prostate structure and serum testosterone levels after chronic treatment with isoproterenol in adult rats with different androgen status.

The effects of chronic administration of the beta-adrenergic agonist isoproterenol (ISO) (120 microg/kg per day; subcutaneously) on the ventral prostate structure and serum testosterone concentrations were examined in adult rats with different androgen status: intact, intact testosterone-injected (1 mg/rat), surgically and chemically castrated rats. Chemical castration was evoked by an intraperitoneal injection of ethane dimethanesulfonate (EDS) (75 mg/kg). A ventral prostate response was only observed in intact and chemically castrated animals. Stereological analysis revealed atrophic changes in the glandular compartment of the prostate of ISO-treated intact rats, but they were probably the consequence of significantly decreased serum testosterone levels. In addition, in these animals alterations were found in the morphometrical parameters of the ventral prostate blood vessels, their relative and total volumes being increased. In chemically castrated rats, administration of ISO from the day of EDS application partially prevented the postcastrational regression of the ventral prostate without affecting blood testosterone level. However, it seems that the discharge of glandular secretion was attenuated at the same time. These results show that chronic treatment with ISO may have both stimulatory and inhibitory effects on the rat ventral prostate depending on the androgen status of the animals and, accordingly, on the site of ISO-action.

The response of rat adrenal zona fasciculata and zona reticularis to oxytocin treatment.

The response of the adrenal zona fasciculata (ZF) and zona reticularis (ZR) to oxytocin (OT) was examined in male rats using a stereological method. The 10-day administration of 0.25 IU OT/100 g daily caused an enlargement of ZF due to cell hypertrophy associated with striking lipid accumulation. The volume of ZR was decreased, this change being the consequence of the reduced cell population. The possible site of OT action is discussed.

The response of rat adrenal medulla to oxytocin.

Effects of oxytocin (OT) on the adrenal chromaffin tissue of male rats were examined by coupled morphometric and biochemical techniques. Synthetic OT was administered in doses of 0.14 and 0.25 IU/100 g/d during 7 or 10 consecutive days and the effects were followed 1, 24, 72 and 168 hours after the last injection. The function and structure of chromaffin cells were affected by the higher dose of OT only. They caused divergent responses on their amine contents. Adrenaline, noradrenaline and dopamine contents were increased, while serotonin content was decreased. These changes were different in duration and time of incidence. Stereological analysis showed an enhanced number of chromaffin cells and an increase in their total volume. The parallelism between the changes in chromaffin cell number and the catecholamine content strongly suggests a mitogenic effect of the applied OT.

Effects of ethane dimethanesulfonate on the structure of adult male rat adrenal cortex.

The influence of ethane dimethanesulfonate (EDS), a specific toxicant for Leydig cells, on the morphometric characteristic of adult male rat adrenal cortex was examined on day 15 after administration. As expected, the dose of 75 mg kg-1 of EDS produced drastic reduction in serum testosterone levels followed by a decrease in testis and seminal vesicle weight. However, a considerable drop (over 50%) in adrenal weight was also found. Stereological analysis of the adrenal cortex, performed under light microscope, revealed atrophy of all adrenocortical zones. The changes were most prominent in the inner zones. Thus, in the zona fasciculata the number of parenchymal cells was markedly decreased. In the zona reticularis a layer consisting mostly of atrophic parenchymal cells was localised at the border between zona reticularis and zona fasciculata. The remaining small number of cortical cells in this zone displayed a notable hypertrophy. It is concluded that EDS at a dose that destroys Leydig cells has a strong deleterious effect on steroidogenic cells of adult male rat adrenal cortex.

The influence of dexamethasone treatment of pregnant rats on the development of chromaffin tissue in their offspring during the fetal and neonatal period.

The aim of these examinations was to determine the influence of dexamethasone (Dx)-treatment of gravid females, on day 16 of gestation on the development of medullary chromaffin tissue of their fetuses and neonatal offspring. In conducting these investigations we used stereological as well as spectrofluorimetric measurements, in 20-day-old fetuses and 1-, 3-, 5-, 7-, 9-, 11-, 13- and 14-day-old neonatal rats. Single Dx-treatment (1.5 mg/kg bw) of the dams led to a significant decrease in body and adrenal weight of their fetuses and neonatal offspring, and also reduction of the medullary volume and the number of chromaffin cells during the entire period examined as a result of decreased cell proliferation in the fetal and early neonatal period (till the 5th day of age). The proliferative activity of the chromaffin cells was evaluated through the mitotic index after applying the cytostatic vincristine-sulphate. During the second neonatal week the mitotic index showed significantly higher values in comparison with the corresponding controls, which indicates that there is regeneration and recovery of the adrenal gland medulla. Adrenaline content in the adrenal gland tissue of offspring of Dx-treated dams was significantly reduced only on the 1st neonatal day. Thus, the change in blood glucocorticoid level of pregnant females after a single Dx injection during the period critical for development of the hypothalamo-pituitary-adrenal system in fetuses affects the development and kinetics of medullar chromaffin cell division.

Stereologic analysis of ventral prostate of oxytocin-treated rats.

Ventral prostate response to oxytocin (OT) was examined in intact and castrated adult rats. The treatment consisted of 10 daily subcutaneous injections of 0.25 IU OT per 100 g body weight, which for orchiectomized rats began immediately after surgery. OT induced prostatic response only in castrated animals. Stereologic analysis revealed changes in the epithelial component, its total volume and surface being increased. The acinar lumen as well as the glandular weight were also enhanced. A possibility that the ventral prostate is the target organ for OT is discussed.

[The effect of gender on vasomotor function of the vascular endothelium and cardiovascular remodelling during aging]. / Uticaj pola na vazomotornu funkciju endotela krvnih sudova i kardiovaskularno remodelovanje tokom starenja.

INTRODUCTION: Aging is correlated with decreased endothelial vasomotor influence, increased carotid intima-media thickness and stiffness, increased left ventricular mass index and increased blood pressure [1-3]. However, these changes are not expressed in the same way in both genders [4, 5]. It seems that females are more protected from cardiovascular changes during aging compared to males [1, 6]. AIM OF THE STUDY: The aim of the study was to evaluate the influence of gender on brachial vasomotor responses (reactive hyperemia test) as well as the correlation with vascular and cardiac remodelling in healthy volunteers of different ages. MATERIAL AND METHODS: The study was carried out on healthy subjects (n = 66; 37 males, 29 females) of different ages (20-82 years) with no history of cardiovascular diseases and diabetes mellitus. All subjects were normotensive, non-smokers with normal blood lipid and glucose values, were not taking any medication at the time of investigation and were asked to refrain from eating and drinking alcohol, coffee or tea 12 hours before the examination. Subjects were divided in two groups (male and female) and 5 age-related groups according to appropriate decade (20-29, 30-39, 40-49, 50-59, and above 60 years). All subjects underwent regular cardiologic examination, ECG recording and cardiac ultrasound in order to exclude valvular diseases, decreased myocardial contractility and ejection fraction. During the study blood pressure and ECG were recorded continuously. Carotid artery intima-media thickness and brachial artery diastolic internal diameter (mm) and blood flow (ml/min) values were measured continuously using high-resolution ultrasound. Brachial artery parameters were measured in baseline condition, during ischaemia and reactive hyperemia (endothelium-dependent relaxation) and after nitroglycerin administration (endothelium independent relaxation, 2 x 400 micrograms, sublingual) [7, 8]. Brachial ischaemia was induced by inflation of a pneumatic tourniquet placed at the forearm to a pressure of 300 mmHg followed by deflation after 3 min. We analyzed changes in peripheral arteries (changes in brachial artery diastolic diameter and flow during 90 sec after cuff deflation), structural changes of carotid artery, functional and structural changes of the left ventricle (19-11). We used cardiac ultrasound (Hewlett Packard Sonos 2500) with a 2.0-2.5 MHz imaging transducer and a 7.0-MHz linear array transducer for vascular measurements. Demographic and clinical characteristics of subjects are presented in Table 1. All results are expressed as mean and S.E.M. Data analysis was done by linear regression analyses, multivariate test (LSD procedure) and Student's T-test. P values less than 0.05 were considered to be significant. RESULTS: Relative changes in brachial artery diastolic diameter in reactive hyperemia in comparison to aging (with gender distribution) are shown in Graphs 1 and 2. Our study showed decrease in brachial vasodilator response to reactive hyperemia during aging (male p < 0.05, female p < 0.001). Data analysis showed a significant difference between age-related groups above 40 years and groups below 30 years of age (p < 0.001). The analysis of carotid intima-media thickness showed increased values during aging in both genders but without statistical significance (Graph 2). Analysis of relationship between carotid intima-media thickness and aging (by gender) showed a good correlation of these parameters expressed by the following formula: intima-media thickness (cm) = 0.0009 x years of age + 0.0139. ANOVA test for age-related groups showed significant correlation (p < 0.01) between all age-related groups except 30-40 vs. 40-50 year group. Student's T-test showed no significant correlation between genders. The relationship between the left ventricular mass index (LVMI) and aging (with gender distribution) is shown in Graphs 3 and 4. The left ventricular mass index was increased during aging

Altered gonodal hormone level and constant light-induced stress interfere with the response of the adrenal medulla to oxytocin

The effect of oxytocin (0.25 IU/100 g per day) on the adrenal medulla was examined in intact, intact estrogen-treated, castrated and castrated testosterone-treated adult male Wistar rats. Stereological analysis of the gland (N = 5 rats per group) revealed that in intact animals the number of chromaffin cells (x10(3)) was significantly increased after 3-day (saline: 467.6 +/- 27.4; oxytocin: 567.6 +/- 28.9) or 7-day (saline: 486.2 +/- 39.1; oxytocin: 618.7 +/- 36.8) oxytocin administration. During 7 days of recovery after the 7-day treatment, the chromaffin cell number returned to the control level (saline: 491.4 +/- 12.6; oxytocin: 554.4 +/- 28.7). The effect of oxytocin on chromaffin cell number was also observed in rats simultaneously injected with estradiol (0.3 microg/100 g per day) for 10 days (estradiol: 454.3 +/- 32.8; estradiol + oxytocin: 576.1 +/- 25.0), as well as in 10-day castrated rats (saline: 594.7 +/- 22.7; oxytocin: 765.3 +/- 33.1). Testosterone replacement (0.6 mg/100 g per day) abolished the medullary response to oxytocin (testosterone + saline: 528.5 +/- 24.7; testosterone + oxytocin: 620.8 +/- 56.0). There was a 20 percent rise in adrenal dopamine content (from 0.236 +/- 0.015 to 0.283 +/- 0.015 microg per pair of glands; N = 9-12) in intact rats injected with oxytocin for 3 days. Oxytocin had no effect on any of the catecholamine levels in adrenal glands of rats exposed to stress induced by constant lighting. The present data indicate that the proliferative response of chromaffin tissue to oxytocin depends on the gonadal hormone level and the basal activity of the adrenal medulla.