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Responses of plasma "estradiol" and plasma LH to ovariectomy, ovariectomy plus adrenalectomy, and estrogen injection at various ages.

Plasma LH and "estradiol" were measured by radioimmunoassay in female rats 5, 10, 15, 20, 25, 30, 40, and 80 days of age, and the changes in these hormone levels 24 h after ovariectomy, ovariectomy plus adrenalectomy, and estrogen injection were determined. Plasma "estradiol" was high at 5 days of age; it increased further to a peak at 15 days of age, and declined thereafter to the low levels seen in adult rats. At each age, plasma "estradiol" declined about 50% 24 h after ovariectomy, whereas it fell to undetectable levels 24 h after ovariectomy plus adrenalectomy at 20 and 80 days of age. Plasma LH did not increase 24 h following ovariectomy at 5 days of age, but it did at all ages thereafter. The peak increase occurred 24 h after ovariectomy at 15 days of age, and the increment was progressively smaller at older ages. Ovariectomy plus adrenalectomy did not cause a significantly greater increase in plasma LH than ovariectomy alone. Injection of estradiol caused a decrease in plasms LH at all ages except in the animals ovariectomized at 5 days of age. The greatest decrease was seen in animals ovariectomized at 15 days of age, with a progressively smaller negative feedback response as age increased. In addition to providing systematic data on changes in the negative feedback effect of estradiol at various ages, the results indicate that, especially in immature animals, there is a circulating substance which cross-reacts with antibodies to estradiol but which does not appear to be a biologically-active estrogen. It does not come from the ovaries and disappears from the circulation if the adrenals as well as the ovaries are removed.

Adrenal Glands↗

Anterior pituitary estradiol receptors associated with the reinstatement of ovulatory cycles after lactation interruption in the rat.

The participation of adenohypophyseal estradiol receptors in the reinstatement of ovulatory cycles after lactation interruption was investigated. In rats whose pups were removed on day 13 postpartum (LRX), prolactin levels fell as from 1600 h on the same day, estradiol peaked on the morning of day 15 and gonadotropins and prolactin (PRL) surged on the afternoon of day 15. No significant changes in gonadotropins or estradiol levels were observed in rats which remained with their litters (LRP); in these rats daily afternoon surges of PRL were detected. No significant variations in anterior pituitary nuclear or cytosolic estradiol receptors were determined in LRP rats. In the nuclear fraction of LRX rats, an important increase (430.8 +/- 124.9%) in receptor titers was observed on day 15. In these animals a significant increase (34.8 +/- 1.3%) in cytosolic estradiol receptors was observed on day 14, followed by a fall on day 15 (-31.6 +/- 6.6%) in comparison to day 13 levels. The receptor variations observed on day 15 closely resemble estrous cyclic changes determined in adult females. However, an observation which does not resemble those cycle variations is the increase in cytosolic receptors observed on day 14 in LRX rats. This increase may be the consequence of a decrease in dopamine levels induced by pup removal. To our knowledge this is the first time that the involvement of pituitary estradiol receptors in the reinstatement of ovulatory cycles after lactation interruption has been described.

Animals↗

The effect of low dose continuous exposure to estradiol on the estrogen receptor (type I) and nuclear type II sites.

These studies were done to evaluate the temporal relationships between cytoplasmic and nuclear estrogen receptors (type I sites), nuclear type II estrogen binding sites, and uterotropic response (true uterine growth; DNA content) after low dose estrogen administration to adult-ovariectomized rats by beeswax pellet implant. Administration of low dose (2.5 micrograms estradiol-17 beta/pellet) estrogen pellets resulted in a slight accumulation of nuclear type I sites (approximately 1700 sites per cell above ovariectomized controls) and a 2-fold stimulation of nuclear type II estrogen binding sites (approximately 5,000 sites per cell) at short times (1-24 h) after estrogen treatment. By 96 h, nuclear levels of type I sites were not different from control even though nuclear type II sites remained elevated and uterotropic response was maintained. Similar temporal responses were observed after treatment with a higher dose estradiol (20 micrograms/pellet) except that early response (1-24 h) to the hormone was accompanied by a more pronounced accumulation (approximately 3,000 sites per cell above control) of type I sites in the nucleus. By 96 h after the administration of the 20-micrograms estradiol pellet, nuclear type I sites declined to control, however nuclear type II sites continued to increase (approximately 30,000 sites per cell) and maximum uterotropic response was observed. Comparison of the results obtained with these two dose levels of estradiol suggested that at early times (1-24 h) after treatment only a very small quantity (approximately 1,700 sites per cell) of nuclear type I sites is required for uterine growth. Likewise, although nuclear type II sites can be maximally stimulated (approximately 30,000 sites per cell) by higher doses of estrogen (20 micrograms) only 5,000 sites per cell may be necessary for maximal response. Surprisingly, uterine growth was sustained by very low to non-measurable quantities of type I sites in the nucleus (96 h). Regardless of the initial numbers of type I sites translocated to the nucleus (approximately 1,700 sites per cell, 2.5 micrograms estradiol; approximately 3,000 sites per cell, 20 micrograms estradiol), at early times (1-24 h), these levels decline to control approximately 96 h after treatment. These results are not due to a decreased rate of estrogen release from the beeswax pellet implant and the data suggest a down-regulation of nuclear type I sites is a normal response to the hormone under these experimental conditions, even though maximum uterine growth is maintained.

Animals↗

Normalization of estradiol receptor kinetics and hormonal activity in uterus of streptozotocin-induced diabetic rats treated with insulin.

In response to ip 17 beta-estradiol injection, ovariectomized streptozotocin-induced diabetic rats showed a decreased early and late stimulation of uterine protein synthesis as compared to nondiabetic controls. An early nuclear loss of estradiol-receptor complex and an accelerated estradiol clearance from the circulating blood were also observed in these animals. To establish whether these alterations in hormone kinetics and activity were related to the diabetic state, further investigation was done by treating diabetic animals with insulin, during 18 consecutive days after 3 to 8 months of long standing diabetes (type I treatment) or during 10 weeks starting immediately at the onset of the streptozotocin-induced disease (type II treatment). Both types of treatment either restored (type I) or preserved (type II) a normal body weight, low glycemia, and absence of glycosuria. Both types of treatment also restored a normal hormonal clearance so that higher plasma levels of estradiol at 30 and 90 min were noted after ip injection when compared to untreated diabetics. Nuclear retention of the estradiol-receptor complex was significantly higher in treated diabetics at 210 min as compared to the nondiabetic controls. Finally, stimulation of early protein synthesis at 90 min was normalized after both types of treatment. From the above observations it is concluded that insulin treatment restored a normal estrogen metabolism in diabetics, resulting in a prolonged retention of the estradiol-receptor complex in the uterine nuclei; the latter event in turn elicited a normal hormonal activity on protein synthesis. A direct influence of the diabetic state and of insulin treatment on the receptor kinetics and/or on the tissue response to the hormonal stimulus may however not be excluded.

Animals↗

Changes in alpha-estradiol receptor and progesterone receptor expression in the locus coeruleus and preoptic area throughout the rat estrous cycle.

We have previously shown that the locus coeruleus (LC) is essential for triggering surges of LH. Since LC neurons are responsive to estradiol, which induces progesterone receptor (PR) expression, this study aimed to investigate whether LC neurons express the alpha-estradiol receptor (alphaER) and PR as well as comparing such responses to that observed in the preoptic area (POA). Female rats were perfused at 10, 14 and 16 h on each day of the estrous cycle, and a blood sample was collected for estradiol, progesterone and LH measurements. alphaER- and PR immunoreactive (ir) neurons were detected in POA and LC by immunocytochemistry (ICC). Higher plasma estradiol levels were observed on the day of proestrus, when a smaller number of alphaER-ir POA neurons were detected. An increase in the number of alphaER-ir neurons were observed at 16 h of proestrus and estrus. The number of PR-ir neurons increased in POA only at 16 h of proestrus, and remained unchanged during all other days and times. The profile of alphaER-ir and PR-ir neurons in LC changed over the estrous cycle, with a lower expression on metestrus morning and reaching a peak on diestrus afternoon before declining on the day of proestrus. However, on estrus afternoon, alphaER-ir neurons increased, while PR-ir neurons decreased which may be related to the prolactin surge of estrus. These data show that LC neurons express alphaER and PR and seem to be more sensitive to variations in estradiol than POA. Also, the fluctuation in alphaER and PR observed for LC neurons seems to accompany the hormonal events that occur during the estrous cycle. This profile of alphaER and PR expression might be related to the ability of estradiol and progesterone in regulating the activity of LC neurons, which could be associated to the control mechanisms of LH and prolactin release.

Animals↗

Estradiol coupling to human monocyte nitric oxide release is dependent on intracellular calcium transients: evidence for an estrogen surface receptor.

We tested the hypothesis that estrogen acutely stimulates constitutive NO synthase (cNOS) activity in human peripheral monocytes by acting on an estrogen surface receptor. NO release was measured in real time with an amperometric probe. 17beta-estradiol exposure to monocytes stimulated NO release within seconds in a concentration-dependent manner, whereas 17alpha-estradiol had no effect. 17beta-estradiol conjugated to BSA (E2-BSA) also stimulated NO release, suggesting mediation by a membrane surface receptor. Tamoxifen, an estrogen receptor inhibitor, antagonized the action of both 17beta-estradiol and E2-BSA, whereas ICI 182,780, a selective inhibitor of the nuclear estrogen receptor, had no effect. We further showed, using a dual emission microfluorometry in a calcium-free medium, that the 17beta-estradiol-stimulated release of monocyte NO was dependent on the initial stimulation of intracellular calcium transients in a tamoxifen-sensitive process. Leeching out the intracellular calcium stores abolished the effect of 17beta-estradiol on NO release. RT-PCR analysis of RNA obtained from the cells revealed a strong estrogen receptor-alpha amplification signal and a weak beta signal. Taken together, a physiological dose of estrogen acutely stimulates NO release from human monocytes via the activation of an estrogen surface receptor that is coupled to increases in intracellular calcium.

Calcium↗

Effects of two oral contraceptives, containing 30 or 20 microg of ethinyl estradiol in combination with gestodene, on blood coagulation and fibrinolysis in Brazilian women.

OBJECTIVE: Blood coagulation and fibrinolytic variables were evaluated in 46 Brazilian women treated with either of two monophasic oral contraceptives (OC), containing 30 or 20 microg of ethinyl estradiol, and 75 microg of gestodene. METHODS: The effects on procoagulants, anticoagulants, pro-fibrinolytics and antifibrinolytics and fibrin turnover were evaluated after treatment for six consecutive cycles, the impact of reduction of ethinyl estradiol dosage on these effects being assessed. RESULTS: The OC containing 30 microg of ethinyl estradiol significantly increased the activities of factors VIII and X, whereas the one containing 20 microg of ethinyl estradiol caused no changes in clotting factors. Neither treatment altered fibrinogen levels or factor VII, IX or XII activity. There were no changes in antithrombin levels, but treatment with 30 microg ethinyl estradiol increased protein C levels and treatment with 20 microg decreased total protein S levels. Concerning the fibrinolytic parameters, both OCs increased plasminogen activity, whereas no changes in PAI-1, t-PA, alpha-2-antiplasmin or fibrin degradation products were observed. The reduction in ethinyl estradiol dosage from 30 microg to 20 microg eliminated the effects on factors VIII and X. CONCLUSIONS: The results show that the OC studied did not cause sufficient changes to indicate that there may be a correlation between these laboratory alterations and clinical results. The lack of reports concerning the hemostatic effects of OCs on Brazilian women hinders comparison of the present data with those obtained for other ethnic groups, at different geographical locations, and emphasizes the importance of such a study for future epidemiological investigation of the prothrombotic effects of OCs in Brazilian women.

Adult↗

Estradiol-promoted accumulation of receptor in nuclei of porcine endometrium cells. Comparison of the retention of receptor in nuclei during subcellular fractionation of untreated and hormone-treated cells.

Nuclei were isolated from porcine endometrium of castrated pigs either unexposed or exposed to estradiol in vivo by two techniques, one of which included a hypotonic step. Aliquots were analyzed for estradiol content. Receptor was extracted from buffered, Surfynol-stabilized suspensions by either (a) KCl alone, (b) in combination with dithiothreitol, or (c) by dithiothreitol with polypentosanesulfate and addition of KCl. The yields rose from a-->c. The same proportional gains with increasing extractant efficacies were obtained from nuclei of unstimulated and estradiol-treated cells. Receptor recovery with extractant "c" rose linearly over the range of 9-80 x 10(6) nuclei/mL and was independent of the technique used for isolation. Nuclear fractions isolated using steroid-free solutions contained more estrogen receptor than estradiol; the numerical excess in control nuclei persisted in the nuclei of stimulated cells featuring a stoichiometric rise of ligand and receptor contents. The increase of receptor contents in nuclei isolated from hormone-stimulated cells coincided with a decline in the cytoplasmic fractions. An excess of hormone over receptor was seen only when nuclei were isolated from untreated cells with media containing 10 nM estradiol. Our data strengthen earlier notions of an estradiol-promoted receptor translocation into the nucleus and are not compatible with the ligand-filling hypothesis of preexisting nuclear binding sites.

Animals↗

Efficacy of estradiol for the treatment of depressive disorders in perimenopausal women: a double-blind, randomized, placebo-controlled trial.

BACKGROUND: Results of previous studies suggest that estrogen improves somatic and mild depressive symptoms experienced by perimenopausal women. This study investigated the efficacy of 17beta-estradiol for the treatment of clinically significant depressive disorders in endocrinologically confirmed perimenopausal women. METHODS: Perimenopausal women (aged 40-55 years, with irregular menstrual periods and serum concentrations of follicle-stimulating hormone >25 IU/L), meeting criteria for major depressive disorder, dysthymic disorder, or minor depressive disorder, according to DSM-IV, were randomized to receive transdermal patches of 17beta-estradiol (100 microgram) or placebo in a 12-week, double-blind, placebo-controlled study. A 4-week washout period followed the 12-week treatment phase. Outcome measures were the Montgomery-Asberg Depression Rating Scale and Blatt-Kupperman Menopausal Index scores. RESULTS: Fifty women were enrolled in the study; 26 met DSM-IV criteria for major depressive disorder, 11 for dysthymic disorder, and 13 for minor depressive disorder. Remission of depression was observed in 17 (68%) women treated with 17beta-estradiol compared with 5 (20%) in the placebo group (P =.001). Subjects responded similarly to estradiol treatment, regardless of DSM-IV diagnosis. Patients treated with estradiol sustained antidepressant benefit of treatment after the 4-week washout period, although somatic complaints increased in frequency and intensity. Treatment was well tolerated and adverse events were rare in both groups. CONCLUSION: Transdermal estradiol replacement is an effective treatment of depression for perimenopausal women.

Administration, Cutaneous↗

Estradiol and fibulin-1 inhibit motility of human ovarian- and breast-cancer cells induced by fibronectin.

Ovarian-cancer cells are characterized by their ability to invade freely the peritoneal cavity. Estradiol stimulates the proliferation of estrogen-receptor(ER)-positive ovarian-cancer cells, as well as expression of fibulin-1, a fibronectin-binding extracellular matrix protein. Using a modified Boyden-chamber assay, we have evaluated the respective roles of estradiol and fibulin-1 on cell motility, one of the earlier steps of tumor invasion. The effect of estradiol was examined on the random and directional migration of different ER-positive ovarian-cancer cell lines. The effect of fibulin-1 was studied on the motility of the MDA-MB231 breast-cancer cell line, which does not express fibulin-1. We found that when fibronectin (FN) was used as an attractant, estradiol decreased the cell motility of 2 ER-positive ovarian-cancer cell lines, BG-1 and SKOV3, but had no effect on 2 ER-negative cell lines, PEO14 and MDA-MB231. The inhibitory effect of estradiol was not observed when collagen (type 1 or 4) or laminin were used as attractants. Fibulin-1 was found to inhibit haptotactic migration of MDA-MB231 cells to FN in a dose-dependent manner. We conclude that both estradiol and fibulin-1 inhibit cancer cell motility in vitro and therefore have the potential to inhibit tumor invasion.

Breast Neoplasms↗

Neuroprotective effects of estradiol in the adult rat hippocampus: interaction with insulin-like growth factor-I signalling.

We have previously shown that 17-beta-estradiol protects neurons in the dentate gyrus from kainic acid-induced death in vivo. To analyse whether this effect is mediated through estrogen receptors and through cross-talk between steroid and insulin-like growth factor (IGF) systems, we have concomitantly administered antagonists of estrogen receptor (ICI 182,780) or the IGF-I receptor (JB1) with estradiol. In addition, we have also administered IGF-I with or without the estrogen receptor antagonist. JB1 (20 microg/ml), ICI 182,780 (10(-7) M), and IGF-I (100 microg/ml) were delivered into the left lateral ventricle of young ovariectomized rats via an Alzet osmotic minipump (0.5 microl/hr) for 2 weeks. All rats received kainic acid (7 mg/Kg b.w.) or vehicle i.p. injections at day 7 after minipump implant. Also on day 7, rats treated i.c. v.with only ICI 182,780 or JB1 received a single i.p. injection of 17-beta-estradiol (150 microg/rat) or vehicle. On day 14 after minipump implant, the rats were killed, brains processed, and the number of surviving hilar neurons estimated by the optical disector technique. Both IGF-I and estradiol treatments resulted in over 90% survival of hilar neurons. The neuroprotective action of estradiol was blocked by ICI 182,780 and by JB1. Furthermore, IGF-I enhancement of neuronal survival was significantly reduced by ICI 182,780. These results indicate that in this model of hippocampal lesion, the neuroprotective effect of estradiol depends both on estrogen receptors and IGF-I receptors, while the protection exerted by IGF-I depends also on estrogen receptors. In conclusion, an interaction of estrogen receptor and IGF-I receptor signalling may mediate neuroprotection in the adult rat hippocampus.

Animals↗

Interactions between nerve growth factor binding and estradiol in early development of the zebra finch telencephalon.

The zebra finch telencephalon exhibits rapid and substantial development in the first few weeks after hatching. In parallel, the rate of estradiol synthesis is very high in the zebra finch forebrain, and estradiol can have potent neurotrophic effects in specific telencephalic regions, including those that control the learning and production of song. In an attempt to elucidate mechanisms regulating telencephalic development, potentially including a role for the large capacity for estrogen production, (125)I-nerve growth factor (NGF) binding was measured in homogenates of telencephalon from zebra finches age 3, 15, 30, 60, and 120 days. The highest density of low- and high-affinity (125)I-NGF binding sites was observed in 3-day-old finches. Using an aromatase inhibitor, Fadrozole, to reduce estradiol levels in 1 to 4-day-old zebra finches significantly decreased both high- and low-affinity (125)I-NGF binding sites. Conversely, treating adult or 8 to 14-day-old hatchlings with estradiol increased high-affinity (125)I-NGF binding sites. These results are consistent with the hypothesis that estradiol influences the level of NGF receptors, and suggest one mechanism through which the steroid could affect brain development. The data also indicate that estradiol and NGF activity may be important for very early development of the telencephalon.

Age Factors↗

Effects of estradiol and progesterone on accumulation of relaxin- and carbohydrate-containing granules in endometrial gland cells of the guinea pig.

Guinea pigs were spayed and given various regimens of injections of estradiol and progesterone. The following were monitored in each animal: pubic separation (relaxin stimulation), resorption of the vaginal membrane (estrogen priming), and the presence of PAS-positive granules and/or relaxin in endometrial gland cells (EGC). Injections of estradiol alone resulted in resorption of the vaginal membrane, accumulation of PAS-positive granules in EGC of all animals, and accumulation of relaxin in EGC of two of four animals; but they did not cause pubic separation. Progesterone injections did not result in resorption of the vaginal membrane, separation of pubes, or accumulation of PAS-positive granules; however, one of three animals demonstrated a few EGC that contained relaxin. Animals that received both estradiol and progesterone exhibited PAS-positive granules and relaxin in EGC as well as separated pubes, but did not have resorbed vaginal membranes. Upon examination with the electron microscope, EGC from animals that received estradiol alone exhibited remarkable numbers of secretory granules that contained a carbohydrate-rich material. Secretory granules were not prominent in EGC from animals that received progesterone alone. Estradiol and progesterone injections resulted in accumulation of secretory granules in EGC that contained relaxin and a carbohydrate-rich material. The observations that estradiol and progesterone induce relaxin production in EGC support the hypothesis that uterine relaxin plays an important role in pregnancy and/or parturition in the guinea pig.

Animals↗

Endogenous estradiol and risk of dementia in women and men: the Rotterdam Study.

We determined whether higher endogenous estradiol levels were associated with lower risk of dementia in older men and women not using hormonal replacement therapy, using a case-cohort design within the Rotterdam Study, a population-based follow-up study on chronic diseases, including dementia, in 7983 subjects aged 55 years or older, and ongoing since 1990. The analyses were based on a random subcohort of 508 women and 438 men, and on 76 women and 53 men with incident dementia. Cox proportional hazards models with robustly estimated standard errors showed that in women higher levels of total estradiol were associated with higher risk of dementia (age-adjusted hazard ratio per standard deviation increase 1.38; 95% CI 1.04-1.84). Age-adjusted HR's of Alzheimer's disease and vascular dementia associated with higher levels of total estradiol (per SD increase) were 1.24 (95% CI 0.87-1.76) and 2.19 (95% CI 1.22-3.92), respectively. Similar results were observed for bioavailable estradiol. Additional adjustments for potential confounders did not change the results substantially. In men, no clear association was observed between estradiol levels and risk of dementia or its subtypes. The findings do not support the hypothesis that higher levels of endogenous estradiol reduce risk of dementia, neither in women nor in men.

Aged↗

Astrocytes in 17beta-estradiol treated mixed hippocampal cultures show attenuated calcium response to neuronal activity.

Glial cells in the brain are capable of responding to hormonal signals. The ovarian steroid hormone 17beta-estradiol, in addition to its actions on neurons, can directly affect glial cells. Estrogen receptors have been described on both neurons and astrocytes, suggesting a complex interplay between these two in mediating the effects of the hormone. Astrocytes sense and respond to neuronal activity with a rise in intracellular calcium concentration ([Ca(2+)](i)). Using simultaneous electrophysiology and calcium imaging techniques, we monitored neuronal activity evoked astrocyte ([Ca(2+)](i)) changes in mixed hippocampal cultures loaded with fluo-3 AM. Action potential firing in neurons, elicited by injecting depolarizing current pulses, was associated with ([Ca(2+)](i)) elevations in astrocytes, which could be blocked by 200 microM MCPG and also 1 microM TTX. We compared astrocytic ([Ca(2+)](i)) transients in control and 24-hour estradiol treated cultures. The amplitude of the ([Ca(2+)](i)) transient, the number of responsive astrocytes, and the ([Ca(2+)](i)) wave velocity were all significantly reduced in estradiol treated cultures. ([Ca(2+)](i)) rise in astrocytes in response to local application of the metabotropic glutamate receptor (mGluR) agonist t-ACPD was attenuated in estradiol treated cultures, suggesting functional changes in the astrocyte mGluR following 24-h treatment with estradiol. Since astrocytes can modulate synaptic transmission by release of glutamate, the attenuated ([Ca(2+)](i)) response seen following estradiol treatment could have functional consequences on astrocyte-neuron signaling.

Action Potentials↗

Estradiol-17 beta-D-glucuronide (E-17G) cholestasis in perfused rat liver: fate of E-17G and choleretic responses to bile salts.

This study was designed to test the hypothesis that increasing the infusion rate of bile salts could overcome drug-induced cholestasis. Cholestasis was induced by administration of 17.5 mumol/L estradiol-17 beta-D-glucuronide during the infusion of taurocholate, tauroursodeoxycholate or dehydrocholate at 20 nmol/min/gm liver. After 30 min, a bolus of 10 mumol of the bile salts was added to the perfusate, and the infusion rate of each bile salt was increased. Taurocholate at a rate of 62 or 125 nmol/min/gm liver, caused a prompt dose-dependent increase of the depressed bile flow and bile salt excretion. A higher rate of taurocholate infusion (180 nmol/min/gm liver) was less effective than either the 62 or 125 rate in increasing bile flow. Infusion of tauroursodeoxycholate at 250 or 390 nmol/min/gm liver also led to a dose-dependent recovery. Further increase of tauroursodeoxycholate infusion rate of 580 nmol/min/gm liver did not provide any additional recovery in bile flow. Dehydrocholate, at rates of 62 or 125 nmol/min/gm liver, gave only a slight enhancement of bile flow. Both taurocholate and tauroursodeoxycholate caused a marked removal of the estradiol-17 beta-D-glucuronide, which had accumulated in the liver. At lower taurocholate infusion rates, the estradiol-17 beta-D-glucuronide was excreted mainly in the bile. At the highest rate, however, biliary excretion of estradiol-17 beta-D-glucuronide declined significantly, and a marked back-efflux of the estrogen into the perfusate was noted. In contrast, tauroursodeoxycholate led to enhanced biliary estradiol-17 beta-D-glucuronide excretion at all increased tauroursodeoxycholate infusion rates and to only a small increase in back-efflux of estradiol-17 beta-D-glucuronide at the two highest tauroursodeoxycholate infusion rates.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Suppressive effects of estradiol on dimethylnitrosamine-induced fibrosis of the liver in rats.

As a model for the analysis of the fibrosuppressive role of estradiol, hepatic fibrosis was induced in male and female rats by the administration of a single dose of dimethylnitrosamine (DMN). The fibrotic response of the male liver after DMN treatment was significantly stronger than that of the female liver. In the male DMN model, estradiol reduced hepatic mRNA for type I and III procollagens and the tissue inhibitor of metalloproteinase-1 (TIMP-1), as well as deposition of type I and III collagen protein total hepatic collagen and malondialdehyde (MDA), a product of lipid peroxidation. Concomitant administration of a neutralizing antibody against rat estradiol enhanced fibrogenesis, as judged by the same parameters. Ovariectomy in the female model had a fibrogenic effect, inducing the hepatic expression of both types of procollagen and TIMP-1; in addition, the number of alpha-smooth muscle actin (alpha-SMA)-positive cells in the liver increased; estradiol replacement was fibrosuppressive in the castrated-female model. In rat hepatic stellate cells incubated in primary culture with estradiol, cell number, type I collagen production, and alpha-SMA expression were all reduced. These findings suggest that estradiol suppressed the induction of hepatic fibrosis, and may in part underlie the more rapid progression in males of hepatic fibrosis and its complications.

Animals↗

Estradiol modulation of plasminogen activator production in organ cultures of human breast carcinomas: correlation with clinical outcome of anti-estrogen therapy.

We have studied the estradiol sensitivity of primary human breast carcinomas in organ culture in a prospective pilot series of 109 tumors. The effect on plasminogen activator (PA) production was used as the end-point of estrogen action. We found that: (i) All tumors secreted detectable levels of urokinase-type PA (uPA); the level of basal uPA production was markedly heterogeneous but showed a weak association with the level of estrogen receptor positivity (p = 0.049). (ii) Only 23.5% of the tumors secreted tissue-type PA (tPA) in addition to uPA; a higher proportion of these tumors had histological characteristics indicative of good prognosis (18% vs. 3% of tumors secreting only uPA). (iii) Estradiol modulated uPA production and this effect was receptor-mediated. (iv) Responsiveness to estradiol was limited to a subset (25 of 60 or 41.7%) of estrogen and progesterone-receptor-positive tumors. (v) Of 20 evaluable patients with lymph-node and receptor-positive breast cancer who received adjuvant anti-estrogen therapy, 11 were identified as estradiol-sensitive by the in vitro PA assay; of these, 10 had no evidence of disease after a median follow-up period of 3+ years. In contrast, of 9 patients with tumors identified as estradiol-insensitive, 4 developed metastases within 3+ years of follow-up. (vi) Consistent with the previously reported inhibitory effect of corticosteroids on uPA production in organ cultures of human tumors, the basal culture level of uPA produced by tumors from patients receiving corticosteroids at the time of surgery was significantly lower than the level of uPA in the remaining tumors (p = 0.029). Also, tumors from patients receiving thyroid hormone, known to stimulate uPA in vitro, showed a slight trend toward increased production of uPA. These results show that hormone effects on tumor PA production are qualitatively similar in organ culture and in the host. This and the emerging individual correlation between sensitivity to estradiol in vitro, as determined by PA, and the clinical effectiveness of anti-estrogen therapy, underscore the potential usefulness of the organ culture approach.

Breast Neoplasms↗