Hilus cells from human postmenopausal ovaries: gonadotrophin sensitivity, steroid and cyclic AMP production

Strips of ovarian hilar tissue taken from 11 post-menopausal women were examined histologically and found to contain clusters of hilus cells in contiguity with non-myelinated nerve fibers. No other steroidogenically active structures were found. Specimens from these strips were incubated for 30 and 240 minutes in Krebs bicarbonate buffer containing 5.5 mM glucose and 1% bovine serum albumin. The specimens produced measureable amounts of androstenedione, estradiol-17 beta and progesterone in vitro. The major steroid formed was androstenedione, and this pattern of steroidogenesis appeared similar to that of postmenopausal ovarian stroma. However, the amounts of steroids formed were considerably higher than in stromal tissue. This implies that the hilus cells possibly may be of greater importance, qualitatively, than the stroma cells for steroidogenesis in the postmenopausal ovary. Addition of hCG to the incubated specimens elicited increase in both cyclic AMP formation and steroid synthesis, especially of estradiol-17 beta, indicating a preserved responsiveness to gonadotrophin in hilus cells from postmenopasual ovaries.     

Ovarian steroid production in a woman with polycystic ovary syndrome associated with endometrial cancer

A young woman with typical polycystic ovary syndrome (PCO) underwent laparotomy for moderately differentiated endometrial cancer. Specimens from the hyperplastic thecal and stromal tissue of the ovaries were incubated for 2 hours in the presence or absence of hCG, 100 IU/ml. Following incubation the tissue content of cyclic AMP and the amounts of progesterone (P), androstenedione (A), testosterone (T) and estradiol-17 beta (E2) in the incubation medium were analysed. For comparison, thecal cells from normal ovaries of regularly menstruating women were incubated under identical conditions. In vivo, the PCO ovaries secreted several-fold greater amounts of T than normal ovaries. In vitro, the thecal cells were much more active, steroidogenically, than the stromal cells of the PCO ovary. Furthermore, the hyperplastic thecal cells of the PCO ovary produced several-fold greater amounts of androgens, and appeared more sensitive to stimulation with hCG, as compared with thecal cells from normal ovaries. The results indicate that in women with PCO associated with endometrial cancer the hyperplastic thecal cells are a significant site of abnormal androgen production and abnormal sensitivity to gonadotropin.     

Expression of messenger ribonucleic acid encoding steroidogenic enzymes in postmenopausal ovaries

To investigate expression of steroidogenic enzymes involved in androgen and estrogen synthesis in the stroma of postmenopausal ovaries.Ovarian stromal tissue samples were obtained at hysterectomy from postmenopausal women who had hysterectomy and oophorectomy. Tissues were frozen immediately in liquid nitrogen and kept frozen until RNA was extracted. Total RNA was examined by Northern blot analysis using (32)P-labeled cDNA probes encoding human P450 side chain cleavage (P450(SSC)), P450(17alpha), and cytochrome P450 aromatase (P450(arom)). Concentrations of mRNA encoding cytochrome P450(SSC), cytochrome P450 17alpha-hydroxylase (CYP17), and P450(arom) were determined in the ovarian stroma.We detected P450(SSC) mRNA in all postmenopausal ovaries studied. P450(SSC) mRNA was increased threefold in the ovarian stroma of postmenopausal women with endometrial cancer and endometrial hyperplasia. CYP17 mRNA was detected in the ovarian stroma of all women with endometrial cancer. P450(arom) mRNA was not detected in the ovaries studied.Postmenopausal ovaries possess enzymes for initiation of steroidogenesis. Enzymes essential for androgen synthesis were expressed in the ovarian stroma of postmenopausal women with endometrial cancer or hyperplasia.     

Hormone concentrations in the homogenates of ovarian tissue and blood serum in postmenopausal women not using hormone therapy

Ovaries in postmenopausal women synthesize steroids, mostly androgens. Removal of the ovaries after menopause may be reflected by menopausal symptoms and arterial hypertension observed during postoperative period, along with a significantly increased risk of death due to cardiovascular complications. It is not understood if the clinical consequences of gonad removal at different time points after menopause are similar. The aim of this study was to evaluate ovarian steroidogenesis and consequently to define the role of the ovaries in postmenopausal women depending on the time after menopause. Concentrations of hormones were determined in ovarian homogenates and serum of postmenopausal women. This study included 207 postmenopausal women. They were divided into groups depending on the time after menopause. All participants had laparotomic removal of the ovaries. Concentrations of estradiol, testosterone and androstenedione were measured in ovarian homogenate and serum. The study revealed that ovarian homogenate and serum concentrations of estradiol, testosterone and androstenedione were the highest in women up to 5 years after menopause and since then significantly decreased. This study showed that testosterone, androstenedione and estradiol are synthesized in the postmenopausal ovaries. The peak synthesis of these hormones occurs up to 5 years after menopause and significantly decreases thereafter.     

Pathological and endocrinological study of epithelial ovarian tumors in post-menopausal women

We have studied 37 cases of ovarian epithelial tumors in post-menopausal women, histopathologically and endocrinologically. The normal values for androstenedione, estrone, estradiol and progesterone in healthy post-menopausal women were less than 83 ng/dl, 75 pg/ml, 30 pg/ml and 0.6 ng/ml, respectively. The numbers of cases, the serum values for which were higher than normal, were 9 of 16 (androstenedione), 9 of 16 (estrone), 22 of 29 (estradiol) and 14 of 20 (progesterone). The serum levels of these 4 sex hormones in cases with ovarian stromal condensation (stromal cell hyperplasia) were higher than normal in 8 of 11 (androstenedione), 8 of 11 (estrone), 17 of 19 (estradiol) and 9 of 12 (progesterone), whereas those in cases with no stromal condensation were elevated in 1 of 5, 1 of 5, 5 of 8 and 5 of 7, respectively. After complete removal of the tumors, these elevated sex hormone levels dropped to normal. After the dexamethasone suppression test, the suppression rates for cortisol, 17-OHCS and 17-KS were 4 times as great as those of DHEA-S (dehydroepiandrosterone-sulfate), estrone, estradiol and progesterone. 17 beta-estradiol was localized in hyperplastic ovarian stromal cells in all cases with stromal condensation. We concluded that many of the ovarian epithelial tumors produce these sex hormones and that hyperplastic stromal cells are the source an increased amount of serum estradiol.     

Polycystic ovary syndrome: a new ultrasonographic and color Doppler pattern

The aim of this study was to evaluate whether patients with partial bilateral polycystic ovaries show different ovarian and uterine blood flow to those with complete bilateral polycystic ovaries, and to investigate whether there is a correlation between ultrasonographic and hormonal parameters.

Fifteen patients with partial polycystic ovaries and eighteen patients with complete bilateral polycystic ovaries underwent clinical, biochemical, gray-scale and color Doppler ultrasonographic evaluation.

Hormonal (luteinizing hormone (LH), follicle-stimulating hormone (FSH), LH/FSH concentration ratio, estradiol, prolactin, androstenedione, testosterone), clinical (body mass index, Ferriman–Gallwey score), ultrasonographic (ovarian volume, number and distribution of subcapsular follicles, stromal score) and Doppler (uterine artery and intraparenchymal vessels pulsatility index, ovarian stromal vascularization) parameters were evaluated, in the early follicular phase (cycle day 3–5) in oligomenorrheic patients, or at random in amenorrheic patients.

Significantly higher androstenedione plasma levels and LH/FSH concentration ratios were observed in complete bilateral polycystic ovaries. In partial polycystic ovaries, gray-scale and color Doppler ultrasonography showed different features in affected and unaffected areas of the ovary, similar to polycystic and normal ovary appearance respectively.

In conclusion, PCOS does not predetermine a single ultrasonographic or Doppler pattern.     

Endocrine studies of normal and polycystic ovarian tissues in vitro.

The purpose of this study was to compare the steroidogenic potential of the granulosa, theca, and medullary tissues from polycystic and normal ovaries. These ovarian endocrine compartments were isolated from appropriate ovaries and were cultured in vitro for three days in the absence (control) and presence of follicle-stimulating hormone (FSH)/luteinizing hormone (LH) (1 lU/ml), N6,O2-dibutyryladenosine-3':5'-cyclic monophosphoric acid (Bu2cAMP) (10(-2)M), and adrenocorticotropic hormone (ACTH) (1.3 U/ml). After the incubation, steroids in the media were measured by radioimmunoassay. Granulosa cells (10(5) cells per dish) from 4 to 7 mm follicles of normal and polycystic ovaries secreted progesterone spontaneously during the culture period and the production of progesterone was markedly stimulated (between tenfold and thirtyfold) by gonadotropins and Bu2cAMP but not by ACTH. Little, if any, androgen (androstenedione, dehydroepiandrosterone, and testosterone) or estrogen (estrone and estradiol) accumulated in the media of any granulosa cell culture. The control cultures of theca tissue from normal and polycystic ovaries secreted large amounts of androstenedione and progesterone and the production of these steroids by normal and polycystic ovary theca was stimulated in most cases by LH/FSH and Bu2cAMP but not by ACTH. Both normal and polycystic ovary theca secreted some testosterone and dehydroepiandrosterone but little, if any, estrone or estradiol accumulated in any theca culture. The medullary tissue of normal and polycystic ovaries produced only trace amounts of steroids in vitro except for the results from one polycystic ovary with hyperthecosis in which case significant quantities of C19 and C18 steroids were secreted. These experiments have demonstrated that isolated granulosa and theca cells from midantral follicles of normal and polycystic ovaries have a similar capacity to secrete C21 and C19 steroids in the absence and presence of trophic agents. Therefore, it seems probable that chronic anovulation in patients with polycystic ovaries is not caused by an obvious deficiency in the de novo steroidogenic potential of the multiple midantral follicles of the polycystic ovaries or by the absence of gonadotropin receptors on the polycystic ovary follicular cells.     

Polycystic ovary syndrome: a new ultrasonographic and color Doppler pattern.

The aim of this study was to evaluate whether patients with partial bilateral polycystic ovaries show different ovarian and uterine blood flow to those with complete bilateral polycystic ovaries, and to investigate whether there is a correlation between ultrasonographic and hormonal parameters. Fifteen patients with partial polycystic ovaries and eighteen patients with complete bilateral polycystic ovaries underwent clinical, biochemical, gray-scale and color Doppler ultrasonographic evaluation. Hormonal (luteinizing hormone (LH), follicle-stimulating hormone (FSH), LH/FSH concentration ratio, estradiol, prolactin, androstenedione, testosterone), clinical (body mass index, Ferriman-Gallwey score), ultrasonographic (ovarian volume, number and distribution of subcapsular follicles, stromal score) and Doppler (uterine artery and intraparenchymal vessels pulsatility index, ovarian stromal vascularization) parameters were evaluated, in the early follicular phase (cycle day 3-5) in oligomenorrheic patients, or at random in amenorrheic patients. Significantly higher androstenedione plasma levels and LH/FSH concentration ratios were observed in complete bilateral polycystic ovaries. In partial polycystic ovaries, gray-scale and color Doppler ultrasonography showed different features in affected and unaffected areas of the ovary, similar to polycystic and normal ovary appearance respectively. In conclusion, PCOS does not predetermine a single ultrasonographic or Doppler pattern.     

Effect of prenatal treatment with ethinyl estradiol on the mouse uterus and ovary

Oral ethinyl estradiol (0.01 or 0.02 mg. per kilogram) administered prenatally to ICR-ICL strain mice before the development of the uterine and ovarian blastermata induced cystic glandular hyperplasia with epidermization in the endometrium and excessive formation of degenerating follicles in the ovaries at 10 to 14 weeks of age. To the evidence for the production by estrogen of malignant uterine neoplasms in rodents and the clinical data showing that cystic glandular hyperplasia develops after prolonged estrogen administration in postmenopausal women and adenocarcinoma in premenopausal or menopausal women using oral contraceptives including ethinyl estradiol, we add the present results which suggest that ethinyl estradiol administered transplacentally can cause malignant transformation of the uterine endometrium in mice.     

Virilization due to ovarian hyperthecosis in a postmenopausal woman

A 51-year-old woman presented with hirsutism and virilization of gradual onset. The serum gonadotropin concentrations were in the postmenopausal range, the serum testosterone concentration was markedly elevated (9.8 nmol/l) and the serum estradiol concentration (220 pmol/l) was elevated above the postmenopausal range. A selective venous catheterization study demonstrated raised serum testosterone and androstenedione levels in ovarian veins and suggested the presence of a left ovarian tumor. The raised peripheral estradiol level was shown to be due to ovarian hypersecretion. After bilateral oophorectomy the serum testosterone became normal. Ovarian histology revealed bilateral stromal hyperthecosis. Ovarian hyperthecosis is a rare but important cause of serum testosterone levels in the neoplastic range. This is the third case reported of postmenopausal virilization due to ovarian hyperthecosis and the first report of a selective venous catheterization study in such a patient.     

Secretion of androgens and estrogens by normal and neoplastic ovaries in postmenopausal women

Peripheral and ovarian venous concentrations of testosterone, androstenedione, estradiol-17 beta, and estrone were measured in 30 postmenopausal women. Ovarian venous concentrations of testosterone and estradiol-17 beta were 5.6 and 4.3 times the peripheral concentrations of women with normal ovaries. This suggests that the ovaries of postmenopausal women continue to secrete these two steroids. Mean peripheral testosterone concentrations in 14 postmenopausal women with a nonfunctional or functional ovarian tumor were increased significantly. Ovarian venous concentrations of these four steroids from the side draining the tumor-bearing ovary were increased in 40 to 80% of the women. The venous concentrations from the normal ovary were also increased in 22% or more of the women with an ovarian tumor. Any ovarian neoplasm in postmenopausal women may be associated with increased ovarian sex-steroid secretion.     

Expression of Syndecan-1 in histologically normal breast tissue from postmenopausal women with breast cancer according to mammographic density.

OBJECTIVE: To analyze the expression of Syndecan-1 in dense and non-dense human breast tissue. METHODS: Specimens of histologically normal tissue were obtained from postmenopausal women undergoing surgery for breast cancer. Each tissue block was subject to radiological examination and pair-wise samples of dense and non-dense tissue were collected. Semi-quantitative assessment of immunohistochemical staining intensity for Syndecan-1 and estrogen receptor subtypes was performed. RESULTS: The expression of Syndecan-1 in all tissue compartments was significantly higher in dense than in non-dense specimens. The strongest staining was recorded in stromal tissue. There was a strong correlation between epithelial estrogen receptor alpha and stromal cell Syndecan-1 expression in dense tissue (rs = 0.7; p = 0.02). This association was absent in non-dense tissue. CONCLUSION: An increase of Syndecan-1 in all tissue compartments and a redistribution from epithelium to stroma may be a characteristic feature for dense breast tissue.     

Granulosa cell tumor of the ovary with Sertoli-Leydig cell tumor components (gynandroblastoma)

A rare granulosa cell tumor with Sertoli-Leydig cell tumor components (gynandroblastoma) arising in the left ovary was reported in a 63-year-old female. Microscopically, the tumor was composed predominantly of granulosa cells arranged mainly in a diffuse solid pattern, but in some areas there were a trabecular pattern and thecofibromatous stromal components. Also, well-differentiated Sertoli-Leydig cell tumor elements were present as a minor component. The tumor produced, endocrinologically, a large amount of estradiol, some androstenedione and a small amount of testosterone. The possibility that estradiol in the present tumor was produced predominantly from androstenedione via estrone was suggested by the results of an in vitro biosynthetic study.     

An in vitro perfusion method for metabolic studies on human postmenopausal ovaries

Five ovaries from four postmenopausal women undergoing bilateral oophorectomy for benign conditions were prepared for vascular perfusion for 1 to 5 hours in a recirculating system containing oxygenized (5% CO2 + 95% O2) synthethic bicarbonate buffered medium (Medium 199 + 4% bovine serum albumin + antibiotics). Two histologically normal ovaries released measurable amounts of progesterone into the medium. One ovary, containing a simple cyst, released progesterone into the perfusate, and this release was further increased after the addition of hCG at a concentration of 10 IU/ml. A perfused cystadenofibroma released cAMP, progesterone, estradiol and testosterone into the perfusion medium with an augmented release of cAMP, estradiol and testosterone after the addition of hCG. hCG also had a stimulating effect on the release of prostanoids from this tumour. Fluid from tumour cysts, obtained prior to perfusion, contained measurable levels of gonadotrophins, steroids and prostanoids. The isolated perfused ovary model may provide a tool for further exploration of endocrine and oncological aspects of the postmenopausal ovary.     

Aromatase in aging women

Cessation of ovarian estrogen secretion is the key event during the climacteric. An enzyme termed aromatase in a number of human tissues and cells, including ovarian granulosa cells, the placental syncytiotrophoblast, adipose and skin fibroblasts, bone, and the brain, catalyzes the conversion of C19 steroids to estrogens. Aromatase expression in adipose tissue and possibly the skin primarily accounts for the extraglandular (peripheral) formation of estrogen and increases as a function of body weight and advancing age. Sufficient circulating levels of the biologically active estrogen, estradiol, can be produced as a result of extraglandular aromatization of androstenedione to estrone, which is subsequently reduced to estradiol in peripheral tissues, to cause uterine bleeding and endometrial hyperplasia and cancer in obese anovulatory or postmenopausal women. Extraglandular aromatase expression in adipose tissue and skin (via increasing circulating levels of estradiol) and bone (via increasing local estrogen concentrations) is of paramount importance in slowing the rate of postmenopausal bone loss. Moreover, excessive or inappropriate aromatase expression was demonstrated in adipose fibroblasts surrounding a breast carcinoma, endometriosis-derived stromal cells, and stromal cells in endometrial cancer and gave rise to increased local estrogen concentrations in these tissues. Whether systemically delivered or locally produced, elevated estrogen levels promote the growth of these steroid-responsive tissues. Finally, local estrogen biosynthesis by aromatase activity in the brain may be important in the regulation of various cognitive and hypothalamic functions. The regulation of aromatase expression in human cells via alternatively used promoters, which can be activated or inhibited by various hormones, increases the complexity of estrogen biosynthesis in the human body. Aromatase expression is under the control of the classically located proximal promoter II in the ovary and a far distal promoter I.1 (40 kb upstream of the translation initiation site) in the placenta. In adipose tissue, two other promoters (I.4 and I.3) located between I.1 and II are used in addition to the ovarian-type promoter II. To add a further twist, promoter use in adipose fibroblasts switches between promoters II/I.3 and I.4 upon treatment of these cells with prostaglandin E2 (PGE2) versus glucocorticoids plus cytokines. Moreover, the presence of a carcinoma in breast adipose tissue causes a switch of promoter use from I.4 to II/I.3. Molecular and cellular mechanisms responsible for estrogen formation and their physiologic and clinical relevance will be reviewed in this article.     

Follicular fluid steroid levels and ovarian steroid secretion in polycystic ovarian disease

Comparison of the plasma levels of steroids between ovarian and peripheral venous blood in polycystic ovarian disease revealed that polycystic ovaries secreted androstenedione, testosterone, dihydrotestosterone, dehydroepiandrosterone and 17-hydroxyprogesterone significantly more than healthy-ovaries at the early follicular phase of the cycle, whereas the levels of estradiol and progesterone did not differ from the control values. Follicular cyst fluid from polycystic ovaries showed significantly higher concentrations of testosterone, androstenedione and dehydroepiandrosterone and much lower levels of estrone, estradiol. progesterone and 17-hydroxyprogesterone than follicular fluid from healthy preovulatory follicles. Testosterone and estradiol did not. however, show any significant difference when follicles in polycystic ovaries were compared to those in healthy ovaries at early follicular phase of the cycle. According to these findings follicle development in polycystic ovaries is arrested at a stage corresponding to the early follicular phase of the cycle. Increased androgen and 17-hydroxyprogesterone secretion by polycystic ovaries seems to originate mainly from the hyperplastic theca interna cells.     

Behavior of some steroids in endometrial adenomatous and atypical hyperplasia

Endometrial adenomatous and atypical hyperplasia are the histopathological pictures that more frequently may develop into an adenocarcinoma. It is believed that a hormonal action may favour the genesis of endometrial cancer. We studied the plasmatic levels of some steroids, considered to be responsible for neoplastic changes, in patients with adenomatous and atypical hyperplasia. With this object we measured plasmatic levels of estrone, estradiol, androstenedione, DHEA-S and testosterone in postmenopausal patients with adenomatous and atypical hyperplasia and in fertile women, both in proliferative and secretory phases. We didn't find any difference in the steroid pattern in the two groups.     

The relation of fertility and ovarian histology after bilateral ovarian wedge resection

The immediate and long term fertility after bilateral ovarian wedge resection in 53 women with clomiphene citrate-resistant hyperandrogenic chronic anovulation is related to the histology of the ovarian wedges. Patients with polycystic ovaries and hyperplastic stromal abnormalities had most spontaneous conceptions and a normal fertility during follow-up. Apparently, chronic anovulation in these cases had been caused by ovarian disease in the face of normal hypothalamic function. Patients with polycystic ovaries without stromal abnormalities often needed postoperative stimulation of ovulation in order to conceive, which may indicate hypothalamic involvement. Patients with large ovaries, normal stroma, and small follicles, who as a group had the lowest serum levels of luteinizing hormone, and patients whose ovaries contained large follicles and cysts without theca cell activity did not benefit from the bilateral ovarian wedge resection. Generally, their postoperative response to medical induction of ovulation did not improve either. Measures to prevent adhesions were not completely successful. Nevertheless, our results suggest that anovulation rather than formation of adhesions causes persistent infertility after bilateral ovarian wedge resection.     

Comparison of the patterns of androstenedione, progesterone, and estradiol during the human menstrual cycle

The pattern of androstenedione (A), serum estradiol (E2), and progesterone was measured during the menstrual cycle. No clear-cut pattern was observed for A such as was observed for E2 and progesterone. The mean levels of A tended to increase from Day 1 of menses to midcycle in all women, following the luteinizing hormone (LH) peak, but with marked fluctuation. A significant decline (p.05) in mean A levels from the LH peak to the day prior to menses was noted in all subjects. The ovaries secreted A throughout the menstrual cycle, with the lowest rate just prior to menses. The marked daily variation in A was most likely caused by the adrenal gland. Whether or not A has a luteal origin has not been resolved.     

Characterization of normal and polycystic ovaries using three-dimensional power Doppler ultrasonography

Purpose : To evaluate the characteristics of polycystic compared to normal ovaries using three-dimensional (3-D) power Doppler ultrasonography. Methods : We recruited 42 volunteers, all of whom were commencing IVF treatment. Each patient was examined in the cycle preceeding the start of drug therapy during the late follicular phase. If eight or more subcapsular follicles of 2–8 mm in diameter in one two-dimensional (2-D) plane were detected in either of the ovaries, the patient was categorized as having polycystic ovaries (PCO); otherwise the ovaries were considered normal. The parameters examined were volume of the ovary, vascularization index (VI), flow index (FI), vascularization flow index (VFI), and mean greyness (MG). In addition, the ovary was arbitrarily divided into cortex and stroma, and thereafter volume, VI, FI, VFI, and MG were calculated for these two regions. Results : Twenty-eight women had normal ovaries and 14 had PCO. The comparison between normal and PCO showed that as a group the PCO were larger, without any differences in VI, FI, VFI, or MG. In patients with PCO, the right ovary was larger than the left one. In patients with normal ovaries, FI was higher on the left side. Division into cortex and stroma revealed that there were no differences in cortical or stromal VI, FI, VFI, or MG between normal and PCO on either side. Conclusions : The ovaries defined as polycystic were larger than normal ovaries, but there was no difference in the echogenicity of the stroma between polycystic and normal ovaries. We were also unable to demonstrate that the polycystic ovarian stroma was more vascularized than the stroma in the normal ovaries.