J Rheumatol. 2001 Dec;28(12):2644-9.
Estrogen increases CD40 ligand expression in T cells from women with systemic lupus erythematosus.

Rider V, Jones S, Evans M, Bassiri H, Afsar Z, Abdou NI.

School of Biological Sciences, University of Missouri-Kansas City, USA. VRideittstate.edu

OBJECTIVE: To examine the in vitro effects of estrogen on CD40 ligand (CD40L) expression in peripheral blood T cells isolated from patients with systemic lupus erythematosus (SLE) and normal controls. METHODS: T cells from female patients with SLE and controls were cultured in serum-free medium without and with 2-fluoroestradiol. Some T cells were activated by further culture on anti-CD3 coated plates. Calcineurin was activated in some T cells by culture in ionomycin. Cell surface CD40L was quantitated by FACS analysis. mRNA expression was measured using semiquantitative PCR. RESULTS: Lupus T cells cultured in medium containing 2-fluoroestradiol showed a significant (p = 0.04) increase in the amount of CD40L on the cell surface, but not in the number of positive cells, compared to the same T cells cultured without estradiol. Estradiol did not significantly change CD40L expression on the surface of T cells from normal women. In addition, the difference in cell surface CD40L between T cells cultured without and with estradiol was significantly greater (p = 0.048) on SLE than on normal T cells. Culture of SLE T cells in medium containing 2-fluoroestradiol followed by T cell receptor (TCR) activation for 2 h using anti-CD3 resulted in a significant (p = 0.04) estrogen dependent increase in CD40L mRNA. The estrogen dependent increases in SLE T cell CD40L mRNA and cell surface protein were blocked by the estrogen receptor antagonist ICI 182,780. SLE and normal T cells pretreated with estradiol and cultured with ionomycin for 2 h to activate calcineurin showed no significant differences in CD40L mRNA. CONCLUSION: These results suggest that estradiol, working through the estrogen receptor, stimulates the expression




Eur J Gynaecol Oncol. 2001;22(5):331-5.
The effect of medroxyprogesterone acetate and norethisterone on the estradiol stimulated proliferation in MCF-7 cells: comparison of continuous combined versus sequential combined estradiol/progestin treatment.

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

Centre for Endocrinology and Menopause, Department of Obstetrics and Gynaecology, University of Tuebingen, Tuebingen, Germany.

OBJECTIVE: Little is known on the type of progestin and regimen type in relation to breast cancer risk. We have compared the effect of medroxyprogesterone acetate (MPA) and norethisterone (NET) on the estradiol stimulated proliferation in MCF-7 cells with respect to different regimens used in combined hormone replacement therapy (HRT). DESIGN: To approximate the in vivo conditions in HRT, MCF-7 cultures were pretreated with estradiol followed by estradiol/progestin treatment to represent the sequential combined model and compared with non pretreated cultures followed by estradiol/progestin treatment for the continuous combined model. RESULTS: When using progestins in the continuous combined form with estradiol (10(-10) M) both progestins showed a significant reduction in the estradiol stimulated proliferation of the MCF-7 cells. In the sequential combined model the addition of MPA led to a stronger significant reduction of MCF-7 proliferation but in a narrower concentration range (from 10(-8) to 10(-6) M) compared to the continuous treatment. NET did not show any significant effect on proliferation in the SC model. CONCLUSION: Different regimen types and different progestins do lead to significantly different effects on the proliferation of a breast cancer cell line. These findings might be useful in the elucidation of potential mechanisms involved in the clinical situation.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11766732&dopt=Abstract estradiol




Metab Brain Dis. 2001 Dec;16(3-4):187-98.
17Beta-estradiol attenuates quinolinic acid insult in the rat hippocampus.

Heron P, Daya S.

Division of Pharmacology, Faculty of Pharmacy, Rhodes University, Grahamstown, South Africa.

A number of studies have shown that 17beta-estradiol has neuroprotective properties. In this study the neuroprotective effect of 17beta-estradiol against quinolinic-acid-induced neuronal damage was investigated. Ovariectomized rats were separated into three groups of five animals each. Rats received daily subcutaneous injections of either olive oil or 17beta-estradiol in olive oil for 7 days prior to and following a single intrahippocampal injection of 1 micromol quinolinic acid in 2 microL phosphate-buffered saline. The brains were removed and the hippocampi either sectioned and stained for microscopic examination or used in glutamate receptor saturation binding studies. Glutamate receptor displacement binding studies were also performed using concentrations of 0.05 nM-5 microM 17beta-estradiol or quinolinic acid. The results show that 17beta-estradiol protects hippocampal neurons from quinolinic-acid-induced neurodegeneration by competing with quinolinic acid to bind to the N-methyl-D-aspartate (NMDA) receptor. This would result in a decrease in intracellular free-calcium influx and resultant neuronal swelling.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11769331&dopt=Abstract estradiol




Zhonghua Fu Chan Ke Za Zhi. 2000 Mar;35(3):172-4.
[Influence of estrogen on the epithelial ovarian cancer cell lines in vitro]

[Article in Chinese]

Yang Y, Shen K, Lang J.

Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, China.

OBJECTIVE: To study the effect of 17-beta estradiol on the growth of the epithelial ovarian carcinoma cell lines in vitro. METHODS: The proliferative capacity of the ovarian carcinoma cell lines CAOV3 and OVCAR3 in the culture medium with 17-beta estradiol was evaluated by the means of the mircoculture tetrazolium assay (MTT) and cell kinetics was analyzed by flow cytometry (FCM). The expression of estrogen receptor (ER) of the two cell lines was examined by the immunohistochemical staining. RESULTS: The growth of both two epithelial ovarian cancer cell lines was slightly inhibited by 17-beta estradiol (0.25-5.00 nmol/L), and the inhibitive rate was 6.3%-36.0%. FCM showed that the kinetics of the two cell lines were obviously changed. Comparing with the kinetics of CAOV3 without estradiol, the cell rate of Stage G0/G1 of CAOV3 with estradiol decreased from 55.0% to 19.0%-30.0%, cell rate of Stage S increased from 30.0% to 52.0%, cell rate of Stage G2/M increased from 14.0% to 17.0%-28.0%; In OVCAR3, cell rate of Stage C2/M decreased from 37% to 22.0%-30.0%, cell rate of Stage S increased from 31.0% to 38.0%-51.0%, cell rate of Stage C0/G1 did not changed much (cell rate without estradiol was 32.0%, while cell rate with estradiol 29.0%-31.0%). Estrogen receptors of the two cell lines were positive and not changed after cultured in the medium with 17-beta estradiol. CONCLUSION: The proliferate capacity of the two epithelial ovarian cancer cell lines CAOV3 and OVCAR3 can be inhibited by the estrogen (0.25-5.00 nmol/L), possibly through the change of the tumor cell kinetics.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11775899&dopt=Abstract estradiol




Chin Med J (Engl). 2000 Jul;113(7):632-5.
Immunolocalization of uterine luminal fluid protein (ULF-250) in rat uterus.

Li X, Zhu L, Koide SS, Yu H.

Department of Pharmacology, Beijing Medical University, Beijing 100083, China.

OBJECTIVE: To access the production of a novel uterine luminal fluid 250 kD protein (ULF-250) during the various phases of the estrous cycle in relation to estrogen concentration, and to validate that the production and secretion of ULF-250 are regulated by estradiol and progesterone. METHODS: An immunohistochemical method was used to localize ULF-250 in rat uteri during each phase of the estrous cycle, and in uteri of ovariectomized rats treated with estradiol and progesterone. RESULTS: Positive immunostaining of ULF-250 occurred in the epithelial cells of the uterus at all phases of the estrous cycle; whereas the stroma was immunonegative. During the proestrus phase of the cycle, the glandular epithelial cells and glandular luminal content were stained strongly. During the estrus phase of the cycle, intense staining occurred in the glandular and uterine luminal epithelial cells, including the luminal content of the glands. In the metestrus phase of the cycle, only uterine epithelial cells were stained; during the diestrus phase, intense staining of the secreted contents of the uterine cavity and the glandular lumen occurred. The distribution of ULF-250 in the uteri of ovariectomized female rats treated with estradiol alone and estradiol plus progesterone were examined. In both groups, intense staining of the glandular luminal epithelial cells of the uterine endometrium occurred; in the estradiol-treated animals, only the luminal contents were stained. The present findings suggest that progesterone inhibits the secretion of ULF-250 that is stimulated by estradiol. CONCLUSIONS: ULF-250 is produced by the glandular and luminal epithelial cells of the uterine endometrium and fluctuates with the phases of the estrous cycle. Its production is stimulated by estradiol and its




J Biol Chem. 2002 Mar 15;277(11):9387-94. Epub 2002 Jan 02.
17beta -Estradiol modulates mechanical strain-induced MAPK activation in mesangial cells.

Krepinsky J, Ingram AJ, James L, Ly H, Thai K, Cattran DC, Miller JA, Scholey JW.

Department of Medicine, University of Toronto, Toronto, Ontario M5G 2C4, Canada. joan.krepinsktoronto.ca

Gender is an important determinant of clinical outcome across a broad spectrum of kidney diseases, but the mechanism(s) responsible for the protective effect of female gender have not been fully elucidated. Remnant kidney glomerular injury is limited in female rats compared with male rats despite similar elevations in glomerular capillary pressure. In vitro, mechanical strain leads to the activation of p44/42 mitogen-activated kinase (p44/42 MAPK) and Jun N-terminal kinase/stress-activated protein kinase (SAPK) in glomerular mesangial cells (MC). Accordingly, we studied the effect of 17beta-estradiol on mechanical strain-induced signal transduction in MC. Exposure of MC to mechanical strain increased p44/42 MAPK activation (3-fold) and SAPK activation (2.5-fold), and kinase activation was inhibited by pretreatment with 17beta-estradiol (10(minus sign8) to 10(minus sign11) m) for 24 h in a dose-dependent manner. Mechanical strain-induced nuclear translocation of p44/42 MAPK and SAPK and nuclear protein binding to AP-1 were also attenuated by 17beta-estradiol. The inhibitory effects of 17beta-estradiol were not reproduced by the cell-impermeable estrogen, BSA/17beta-estradiol, nor did preincubation with 17beta-estradiol lead to actin cytoskeleton disassembly or impaired stress fiber formation. However, 17beta-estradiol did increase base-line levels of the dual specificity phosphatase MKP-1. The inhibitory effects of 17beta-estradiol on p44/42 MAPK activation and SAPK activation, translocation, and AP-1 binding were all abrogated by the estrogen receptor antagonist, ICI-182,780. We conclude that attenuation of mechanical strain-induced MAPK activation by 17beta-estradiol is depen




Zhonghua Fu Chan Ke Za Zhi. 2001 Feb;36(2):95-7.
[Effects of leptin on estradiol and progesterone production by human luteinized granulosa cells in vitro]

[Article in Chinese]

Guo X, Chen S, Xing F.

Department of Genocology and Obstetrics, General Hospital of Guangzhou Military District, Guangzhou 510010, China.

OBJECTIVE: To investigate the effects of leptin on steroidogenesis of human luteinized granulosa cell in vitro. METHODS: Human luteinized granulosa cells were isolated from follicular fluid obtained during oocyte retrieval of in vitro fertilization-embryo transfer program and were cultured with M199 medium plus various concentration of leptin (0, 10, 30, 100, 300 ng/ml), human menopausal gonadotropin (hMG, 0, 0.1, 0.2, 0.5, 1, 2, 5, 10 IU/ml) and testosterone 100 micrograms/ml. For 2 days the media were collected for estradiol and progesterone measurements. RESULTS: Addition of leptin alone did not alter estradiol and progesterone production (P > 0.05) by human luteinized granulosa cells. Leptin of 10-30 ng/ml concentrations caused a dose-dependent inhibition of estradiol production (P < 0.05) while greater than 0.5 IU/ml of hMG were added. There was no effect of leptin on hMG-stimulated progesterone production (P > 0.05). CONCLUSION: Leptin can directly inhibit hMG-stimulated estradiol production by human luteinized granulosa cells in vitro, but has no effect on progesterone production. Leptin may play an important role in follicle development and luteinization.

http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=11783356&dopt=Abstract estradiol




J Reprod Fertil Suppl. 2001;57:269-73.
Efficacy of the GnRH analogue deslorelin for suppression of oestrous cycles in cats.

Munson L, Bauman JE, Asa CS, Jochle W, Trigg TE.

Dept VM-PMI, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.

The aim of this study was to develop a method for long-term but reversible inhibition of oestrous cycles in female cats by downregulation of GnRH receptors with deslorelin released from a long-acting implant. In a blind study with mature cats (n = 20), a 6 mg deslorelin implant was administered s.c. to ten cats and a placebo implant was administered to ten cats. Occurrence of oestrus and general health were observed daily, and individual faecal samples were collected at 3 day intervals for 14 months and analysed for oestradiol content. All the placebo-treated queens continued to undergo normal oestrous cycles during the study. Oestrus was accompanied by peaks in oestradiol concentrations of > or = 20 ng g-1 faeces. Treatment with deslorelin initially stimulated oestradiol release, which accompanied treatment-induced ovulations. Thereafter, oestradiol concentrations decreased to 1-10 ng g-1 faeces and remained low for extended periods. Observations of small increases in oestradiol concentrations in one cat led to a second treatment with 6 mg deslorelin in five cats on day 155 after first treatment. Faecal oestradiol concentrations remained < 20 ng g-1 faeces in the five single treatment cats for 8.0, 8.5, 11.0 and 14.0 (two cats) months. Cats receiving two implants had the first oestradiol peak > 20 ng g-1 faeces after treatment at 7.5, 11.0 (two cats), 11.5 and 14.0 months. After 14 months, two cats had returned to normal cyclic activity, two had irregular small oestrogen peaks and six showed no cyclic activity. For months 2-5, 6-10 and 11-14, oestrogen values in treated cats were significantly different from control values (P < 0.001, 0.05 and 0.02, respectively). Differences in oestrogen concentration between contro




J Reprod Fertil Suppl. 2001;57:407-14.
A model for the study of cystic endometrial hyperplasia in bitches.

Chen YM, Wright PJ, Lee CS.

Department of Veterinary Science, University of Melbourne, 250 Princes Highway, Werribee, Victoria 3030, Australia.

The aims of this study were: (i) to establish a reliable model for the study of cystic endometrial hyperplasia in ovariectomized bitches; and (ii) to assess the roles of oestrogen and progesterone in the pathogenesis of irritant-induced cystic endometrial hyperplasia. Greyhound bitches (n = 15) were ovariectomized and divided into five groups (n = 3 per group). After 3-4 weeks, oestradiol benzoate (0.6-4.8 micrograms kg-1, i.m.) was administered twice a day for 12 days to the bitches in group 1, followed by progesterone (0.2-1.8 mg kg-1, i.m.) twice a day for 30-33 days. These dosages were chosen to mimic the plasma hormone concentrations of a normal oestrous cycle. A silk suture was inserted by laparotomy into the left uterine horn 12 days into the simulated dioestrus (determined by vaginal cytology) and necropsy was performed after a further 12 days. For groups 2-5, the silk suture was positioned at ovariectomy. After a further 3-4 weeks, these bitches were treated with progesterone (group 2: 1.8 mg kg-1 i.m. twice a day), oestradiol benzoate (group 3: 0.6-4.8 micrograms kg-1 i.m. twice a day), oestradiol benzoate and progesterone together (group 4: previous dosages) or vehicle (group 5). Necropsies were performed after 12-13 days of treatment. Cystic endometrial hyperplasia was induced in the suture-containing uterine horns of all bitches in groups 1 and 4, and in two bitches in group 2. Cystic endometrial hyperplasia did not develop in any control (no suture) uterine horns, or in either uterine horn of the bitches treated with either oestradiol only or vehicle. These results indicate that progesterone is necessary for the development of irritant-induced cystic endometrial hyperplasia and that oestradiol potentiates the effects of progester