Clin Pharmacol Ther. 2002 May;71(5):311-24.
Stimulatory effect of cigarette smoking on the 15 alpha-hydroxylation of estradiol by human term placenta.

Zhu BT, Cai MX, Spink DC, Hussain MM, Busch CM, Ranzini AC, Lai YL, Lambert GH, Thomas PE, Conney AH.

Department of Basic Pharmaceutical Sciences, College of Pharmacy, University of South Carolina, Columbia, SC, USA.

OBJECTIVE: Our objective was to characterize the oxidative metabolism of estradiol by human term placenta and its modulation by cigarette smoking. METHODS: Placental microsomes were prepared from term placentas obtained from 13 cigarette smokers (20 to 30 cigarettes per day until the time of delivery) and 13 control subjects who were nonsmokers. Estrogen metabolism was studied by incubation of 250 nmol/L [(3)H]estradiol with placental microsomes and NADPH, and the estrogen metabolites were determined by HPLC and gas chromatography-mass spectrometry. RESULTS: 2-Hydroxyestradiol was the major hydroxyestrogen detected, followed by 6alpha-hydroxyestradiol. Small amounts of several other hydroxyestrogen metabolites (4-hydroxyestradiol, 6beta-hydroxyestradiol, 7alpha-hydroxyestradiol, and 16alpha-hydroxyestradiol) were also detected. Large amounts of estrone plus small amounts of 2-hydroxyestrone and unidentified nonpolar metabolites were formed. Cigarette smoking stimulated the placental hydroxylation of benzo[a ]pyrene by about 16-fold. Cigarette smoking had little or no effect on the overall rate of placental estradiol metabolism or on the formation of estrone, 2-hydroxyestradiol, 2-hydroxyestrone, or 16alpha-hydroxyestradiol. However, placental formation of 4-hydroxyestradiol and 7alpha-hydroxyestradiol was increased 38% (P =.08) and 150% (P =.05), respectively, in cigarette smokers. The formation of 6alpha-hydroxyestradiol was decreased 33% (P =.04). Metabolic formation of 15alpha-hydroxyestradiol was observed during incubations of estradiol with placental microsomes from 11 of the 13 cigarette smokers, but this metabolite was not det




Eur J Gynaecol Oncol. 2002;23(2):127-30.
Tibolone versus 17beta-estradiol/norethisterone: effects on the proliferation of human breast cancer cells.

Lippert C, Seeger H, Wallwiener D, Mueck AO.

Department of Obstetrics and Gynecology, University of Tuebingen, Germany.

Tibolone is a synthetic progestin with estrogenic and progestogenic properties, widely used for alleviation of menopausal syndromes and for osteoporosis prophylaxis in postmenopausal women. Since only little data are available on tibolone and breast cancer risk the present study investigates the effect of tibolone on the growth of the human breast cancer cell line, MCF-7. Tibolone is clinically comparable to an estradiol/norethisterone combination, therefore we included this hormone combination in our experiments. Tibolone was examined alone and in the presence of 0.1 nM estradiol in the concentration range from 0.001 microM to 1 microM. Norethisterone was studied using the same concentration range in combination with 0.1 nM estradiol. Tibolone led to significant cell growth in the concentration range of 0.01 to 1 microM and was able to significantly stimulate estradiol-induced proliferation at the concentrations 0.01 and 0.1 microM. In contrast, the estradiol/norethisterone combination elicited significant inhibition of cell growth at the concentrations 0.001 and 0.01 microM. These data suggest that tibolone does have tumor cell-growth promoting effects in vitro whereas the estradiol/norethisterone combination partially inhibits cell growth. Therefore no differences in risk profile are to be expected between conventional hormone substitution using estradiol and norethisterone acetate and tibolone. Drawing a clinical consequence from our experiments would result in not recommending the use of tibolone in postmenopausal women at high risk for breast cancer development until long-term controlled clinical studies have been performed on the effect of tibolone administration and breast cancer risk.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12013108&dopt=Abstract estradiol [PubMed - in




Ther Umsch. 2002 Apr;59(4):175-81.
[Influence of the ovarian cycle on the central nervous system]

[Article in German]

Kuhl H.

Zentrum der Frauenheilkunde und Geburtshilfe Klinikum der J.W. Goethe-Universitat Frankfurt. H.Kuhm.uni-frankfurt.de

Estradiol, progesterone and some of their metabolites modulate the activity of neurotransmitters and neuropeptides in the CNS. The distribution and concentrations of sex steroids in the various CNS regions is partly dependent on the serum levels, but also on the local synthesis of the steroids. In general, estradiol and testosterone exert a stimulatory, progesterone an inhibitory effect on neuronal activities which are mediated by excitatory (e.g. glutamate, aspartate), and inhibitory amino acids (e.g. GABA) and neuropeptides (e.g. beta-endorphin), respectively. Gonadotropin release is primarily governed by the rhythm of pulsatile secretion of GnRH in the hypothalamus which is controlled by estradiol and progesterone by means of inhibitory or stimulatory modulation of the amplitude and frequency of GnRH pulses. The discharges of GnRH neurons triggered by excitatory amino acids are modulated by estradiol, while the inhibitory effect of progesterone is mediated by GABA and beta-endorphin which cause hyperpolarization of the GnRH neurons and consequently a reduced pulse frequency. The pulse amplitudes are primarily influenced by estradiol, but neuropeptide Y, neurotensin and noradrenaline contribute to their preovulatory enhancement. The postovulatory rise in core temperature is caused by the increasing level of progesterone and its metabolite 3 alpha-pregnanolone, respectively. Despite of this, up to 20% of ovulatory cycles do not show any rise in body temperature. Although 3 alpha-pregnanolone has sedative activities, there is no change in sleep quality during the luteal phase due to their low serum levels. It could be demonstrated that performance on tests of articulatory and fine motor skills are enhanced in the late follicular phase as




Chem Res Toxicol. 2002 May;15(5):754-64.
Reactions of estradiol-2,3-quinone with deoxyribonucleosides: possible insights in the reactivity of estrogen quinones with DNA.

Convert O, Van Aerden C, Debrauwer L, Rathahao E, Molines H, Fournier F, Tabet JC, Paris A.

Laboratoire de Chimie Structurale Organique et Biologique, CNRS UMR7613, UPMC, 4 place Jussieu, 75252 Paris Cedex 05, France. oconvercr.jussieu.fr

Estrogen 2,3- and 3,4-quinones are reactive species toward nucleophiles and Michael acceptors. As such, they can bind to DNA and induce cellular damages. As an alkylation model, reactions of estradiol-2,3-quinone with deoxyribonucleosides were previously studied by mass spectrometry. In this work, estrogen-deoxyribonucleoside adducts were synthesized by reaction of 17beta-estradiol-2,3-quinone with deoxyguanosine or deoxyadenosine and analyzed by NMR and LC-MS(n)() in order to determine the structure and the stereochemistry of the resulting covalent adducts. Although estradiol- and estrone-2,3-quinones were previously thought to give mainly stable adducts, identification of depurinating adducts with both nucleosides, i.e., 2-OHE(2)-6(alpha,beta)-N7Gua and 2-OHE(2)-6(alpha,beta)-N7Ade, was unambiguously obtained. This is of particular interest since depurinating adducts are generated from DNA, and therefore, their amount should be correlated to the parallel formation of apurinic sites, which might play an important role in the cancer initiation process. Besides, a byproduct, i.e., 2-hydroxy-11-oxo-estradiol, corresponding to an unstable alkylation product of 2-hydroxyestradiol has been unambiguously identified and is indicative of a plausible addition process at the C9 position of catechol estrogens. The synthetic adducts will be useful as reference compounds to further elucidate the structure of adducts formed by reaction of estrogen metabolites with DNA or oligonucleotides.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12018999&dopt=Abstract estradiol




Life Sci. 2002 May 24;71(1):15-29.
Effects of testosterone and 17-beta-estradiol on TNF-alpha-induced E-selectin and VCAM-1 expression in endothelial cells. Analysis of the underlying receptor pathways.

Zhang X, Wang LY, Jiang TY, Zhang HP, Dou Y, Zhao JH, Zhao H, Qiao ZD, Qiao JT.

Laboratory of Molecular Biology, Shanxi Medical University, 030001 Taiyuan, Shanxi, PR China.

This study investigated the effects of testosterone and 17-beta-estradiol on tumor necrosis factor-alpha (TNF-alpha)-induced endothelial expression of E-selectin and vascular cell adhesion molecule-1 (VCAM-1) and the potential roles of hormone receptors involved in these actions. Human umbilical vein endothelial cells (HUVEC) were stimulated with TNF-alpha in the presence or absence of testosterone or 17-beta-estradiol, and the expression of E-selectin and VCAM-1 was investigated. As shown by Western blot analysis, co-administration with testosterone or 17-beta-estradiol increased the expression of E-selectin and VCAM-1 induced by TNF-alpha at 6 h and 3 h, respectively. Similarly, RT-PCR analysis revealed a significant increase in the amount of mRNA for E-selectin and VCAM-1 after co-administration with testosterone or 17-beta-estradiol in TNF-alpha-stimulated HUVEC. The presence of mRNA and proteins for androgen receptor and estrogen receptor alpha in HUVEC was verified by RT-PCR and Western blot. Flow cytometric analysis showed that preincubation with androgen receptor antagonist cyproterone and estrogen receptor antagonist tamoxifen completely abrogated the upregulating effects of testosterone and 17-beta-estradiol on TNF-alpha-induced E-selectin and VCAM-1 expression, respectively. Expression of TNF receptors in TNF-alpha-stimulated HUVEC was not influenced by testosterone and 17-beta-estradiol. The data indicate that both testosterone and 17-beta-estradiol increase TNF-alpha-induced E-selectin and VCAM-1 expression in endothelial cells via a receptor-mediated system, and expression of TNF receptors are not chan




Endocrinology. 2003 May;144(5):1876-86.
Evidence for a negative intrafollicular role for inhibin in regulation of estradiol production by granulosa cells.

Jimenez-Krassel F, Winn ME, Burns D, Ireland JL, Ireland JJ.

Molecular Reproductive Endocrinology Laboratory, Department of Animal Science, Michigan State University, East Lansing, Michigan 48824, USA.

Intrafollicular concentrations of inhibin A and estradiol vary inversely during development of dominant follicles in cattle. Thus, we hypothesized that inhibin has a negative autocrine or paracrine effect on estradiol production by granulosa cells. To examine this hypothesis, a homologous model system was used to test the effects of bovine antibovine inhibin antibodies, bovine inhibin, and a peptide fragment of bovine inhibin (bINH) on capacity of granulosa cells isolated from individual estrogen-active or -inactive dominant or subordinate follicles to produce estradiol during short-term (18 h) serum-free culture. Immunoblot analysis of media demonstrated that granulosa cells basally produce different molecular weight forms of inhibin, similar to those in bovine follicular fluid. Immunoneutralization of endogenous inhibin in culture with different doses (12.5-1000 microg) of highly purified bovine antibovine inhibin antibodies increased estradiol production 2- to 15-fold, compared with controls. Preadsorption of the anti-inhibin antibodies with bINH precursors or bovine pro-alpha(C) suppressed the capacity of anti-inhibin antibodies to enhance estradiol production by granulosa cells, compared with controls. Treatment of granulosa cells with an immunoaffinity-purified preparation of bINH suppressed basal estradiol production 60%, compared with controls. In contrast, treatment of granulosa cells with the bINH peptide increased estradiol production 14-fold, compared with controls. Based on these results, we concluded that both anti-inhibin antibodies and bINH blocked the suppressive local effects of basally produced inhibin on estradiol produc




Maturitas. 2002 May 20;42(1):63-9.
Beneficial effects of hormonal replacement therapy on chromium status and glucose and lipid metabolism in postmenopausal women.

Roussel AM, Bureau I, Favier M, Polansky MM, Bryden NA, Anderson RA.

LBSO, Universite Joseph Fourier, Domaine de la Merci, La Tronche, France.

OBJECTIVES: Postmenopausal women exhibit an increased incidence of cardiovascular diseases, and type 2 diabetes mellitus compared with younger women. However, women receiving hormonal replacement therapy (HRT) seem to be protected. Since chromium (Cr) functions in glucose, lipid and corticosteroid metabolism and these variables, as well as Cr status, decline with age, Cr status may be a contributing factor in the effects of hormone replacement therapy. Therefore, the objective of this study was to determine the effects of hormonal replacement therapy (HRT) on serum and urinary Cr, plasma lipids, glucose, fructosamine and the related hormonal variables, estradiol, insulin, leptin, cortisol, and DHEA-sulfate. METHODS: Forty-four healthy postmenopausal women 50-60 years old participated in the study. Eighteen were treated by combined oral hormonal replacement therapy (estradiol 2 mg per day during days 1-25 and 10 mg of dydrogesterone on days 10-25) for at least 2 years, and 26 were untreated. RESULTS: Serum Cr concentrations were significantly lower in untreated postmenopausal women than in women receiving HRT (0.070+/-0.008 vs. 0.100+/-0.008 ng/ml) whereas urinary Cr excretion was increased (0.14+/-0.02 vs. 0.07+/-0.01 ng of Cr/mg creatinine). The urinary losses of Cr were inversely correlated with plasma estradiol. Median value of urinary Cr was higher in postmenopausal women exhibiting endogenous estradiol levels below 250 pmol/l, whereas women with estradiol levels >250 pmol/l, exhibited lower Cr values. Plasma fructosamine, total and LDL cholesterol and TC/HDL ratio, which are all decreased by improved Cr nutrition, were also improved in the women receiving HRT. There were also n



Biol Reprod. 2002 Jun;66(6):1640-8.
Effect of follicle size on in vitro production of steroids and insulin-like growth factor (IGF)-I, IGF-II, and the IGF-binding proteins by equine ovarian granulosa cells.

Davidson TR, Chamberlain CS, Bridges TS, Spicer LJ.

Department of Animal Science, Oklahoma State University, Stillwater, Oklahoma 74078, USA.

Little is known regarding the hormonal regulation of granulosa cell steroidogenesis and the ovarian insulin-like growth factor (IGF) system in the mare. The objectives of this study were to determine, first, if estradiol, insulin, and/or FSH affect steroid production by equine granulosa cells (experiment 1) and, second, if the components of the IGF system are produced by equine granulosa cells in culture as well as whether estradiol, insulin, and/or FSH affects IGF and/or IGF-binding protein (IGFBP) production by equine granulosa cells (experiment 2). Granulosa cells from small (6-15 mm), medium (16-25 mm), and large (25-48 mm) follicles were collected from cyclic mares (n = 14), cultured for 2 days in medium containing 10% fetal calf serum, washed, and then treated for an additional 2 days in serum-free medium with or without added hormones. In experiment 1, large-follicle granulosa cells produced less progesterone and more estradiol than did medium- and/or small-follicle granulosa cells (P < 0.05). Progesterone production was inhibited (P < 0.05) by FSH and insulin in small- and medium- but not in large-follicle granulosa cells; estradiol was without effect. Insulin increased (P < 0.05) estradiol production in small- and medium-follicle granulosa cells but had no effect in large-follicle granulosa cells. In experiment 2, IGF-I production was inhibited (P < 0.05) by insulin across all follicle sizes but was not affected by estradiol or FSH. Granulosa cells of medium and large follicles produced more IGF-II than did granulosa cells of small follicles (P < 0.05). Insulin and FSH inhibited (P < 0.05) IGF-II production by granulo




Biol Reprod. 2002 Jun;66(6):1689-95.
An alteration in the hypothalamic action of estradiol due to lack of progesterone exposure can cause follicular cysts in cattle.

Gumen A, Wiltbank MC.

Department of Dairy Science, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.

Many mammals, including cattle, can develop ovarian follicular cysts, but the physiological mechanisms leading to this condition remain undefined. We hypothesized that follicular cysts can develop because estradiol will induce a GnRH/LH surge on one occasion but progesterone exposure is required before another GnRH/LH surge can be induced by estradiol. In experiment 1, 14 cows were synchronized with an intravaginal progesterone insert (IPI) for 7 days, and prostaglandin F(2alpha) was given on the day of IPI removal. Estradiol benzoate (EB; 5 mg i.m.) was given 3 days before IPI removal to induce atresia of follicles. Cows were given a second EB treatment 1 day after IPI removal to induce a GnRH/LH surge in the absence of an ovulatory follicle. All cows had an LH surge following the second EB treatment, and 10 of 14 cows developed a large-follicle anovulatory condition (LFAC) that resembled follicular cysts. These LFAC cows were given a third EB treatment 15 days later, and none of the cows had an LH surge or ovulation. Cows were then either not treated (control, n = 5) or treated for 7 days with an IPI (n = 5) starting 7 days after the third EB injection. Cows were treated for a fourth time with 5 mg of EB 12 h after IPI removal. All IPI-treated, but no control, cows had an LH surge and ovulated in response to the estradiol challenge. In experiment 2, cows were induced to LFAC as in experiment 1 and were then randomly assigned to one of four treatments 1) IPI + EB, 2) IPI + GnRH (100 microg), 3) control + EB, and 4) control + GnRH. Control and IPI-treated cows had a similar LH surge and ovulation when treated with GnRH. In contrast, only IPI-treated cows had an LH surge following EB treatment. Thus, an initial G