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TNF-α alters estrogen metabolism and homeostasis in endometrial cells
phosphorylation of ERK which in turn inhibits transcription and could promote apoptosis. 2-methoxy estradiol had no effect on vasodilation as there was no change in endothelial nitric oxide synthase expression as compared to control. Supported by: None. P-45 TNF-a ALTERS ESTROGEN METABOLISM AND HOMEOSTASIS IN ENDOMETRIAL CELLS. S. M. Salih, M. Kamel, T. D. Veenstra, A. Concepcion, R. Kumar, S. Salama. Obstetrics and Gynecology, University of Wisconsin, Madison, WI; Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, TX; SAIC-Frederick, INC, Frederick, MD. OBJECTIVE: Perturbed estrogen (E2) hemostasis lead to a variety of women health disorders such as abnormal uterine bleeding, endometrial hyperplasia, and endometrial cancer (EC). In this respect, the in situ estrogen bioavailability in endometrial tissues depends on the expression and activity of estrogen metabolizing enzymes as well the ratios of various endogenous estrogen metabolites. Here we investigate the effect of proinflammatory mediators on estrogen metabolism. DESIGN: The current study was undertaken to test the hypothesis that the transcription factor NF-kB, a hallmark of inflammatory responses, contributes to the development of abnormal estrogen environment by altering the balanced expression of enzymes involved E2 biosynthesis and metabolism in endometrial cells. MATERIALS AND METHODS: We used HPLC/electrospray ionizationtandem mass spectrometry analysis to determine cellular E2/E2 metabolites. Western blot analysis, reporter gene assay, and real time RT-PCR were used to assess the effect of TNF-a on the expression of the key enzymes involved in E2 biosynthesis and metabolism. RESULTS: Our data demonstrated that treatment of endometrial cells TNF-a (5 ng/ml) increased the de novo biosynthesis of estrogen. TNF- a increased in the level of total estrogen/estrogen metabolites by 4-fold compared with treatment with E2 alone. In addition, TNF-a directed the metabolism of into more hormonally active estrogenic metabolites. The rate of conversion of the hormonally less active estrone (E1) into the hormonally potent E2 was significantly increased by treatment with TNF-a. In addition, our data clearly indicated that the TNF-a significantly increased the oxidative metabolism of E2/E1 into carcinogenic metabolites 4-hydroxyestardiol; 16a hydroxyestrone; and 17-epiestriol, with concomitant inhibition of the formation of antiestrogenic metabolites 2-methoxyestradiol. Gene expression analysis revealed that TNF- a induced the expression of the key gene involved in E2 biosynthesis (Steroidogenic factor 1, Aromatase; 17beta-hydroxysteroid dehydrogenase type 1), and bioactivation (cytochrome P450-1B1), with concomitant repression of genes involved in inactivation of estrogen (Catechol-O-methyltransferase, and NADPH-quinone oxidoreduactase). CONCLUSIONS: In conclusion, our study suggested that the inflammatory mediator TNF- a affect estrogen metabolism and homeostasis. This may contribute, at least in part, to the risk of the inflammation-associated increased risk for EC and other endometrial pathology. Supported by: None. P-46 THE EFFECT OF FEMALE AGE ON GRANULOSA/THECA CELL HORMONAL PRODUCTION. A. M. Zamah, M. P. Rosen, S. H. Sohn, M. I. Cedars. Obstetrics, Gynecology and Reproductive Sciences, University of California San Francisco, San Francisco, CA. OBJECTIVE: It is well established that the genetic and metabolic potentials of the oocyte decline with female age. Less is known about how aging affects the function of the supporting somatic cell compartment of the follicle. These somatic cells undergo rapid proliferation, induction of metabolic activity and steroid production during folliculogenesis and corpus luteum formation. Data suggest that with advancing female age, serum levels of sex hormones decline in natural menstrual cycles, and clinically, evidence of luteal phase deficiency becomes more common. Whether these changes are due to decreased numbers of metabolically active follicles, changes in ovarian vascularity, or deficiencies within the granulosa and theca cells themselves remains unclear. We have compared follicular fluid levels of sex hormones from a cohort of women age 35 years or younger and over age 40 undergoing ovarian stimulation for IVF in order to address this question. DESIGN: A retrospective study involving patients undergoing ART by standard ovarian stimulation protocols at an urban academic medical center.
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Abstracts
MATERIALS AND METHODS: 304 patients were divided into two groups by age: Group I age % 35 years and Group II age R 40. A total of 429 individual follicular fluid (FF) aspirates were collected at the time of oocyte retrieval. Each patient had 1 or 2 individual follicle aspirates collected. Concentrations of FSH, hCG, estradiol, progesterone, and testosterone were determined by automated chemiluminescent immunoassays on the DPC-Immulite 2000 (Diagnostic Products, Los Angeles, CA). Linear regression was initially performed to evaluate the difference in hormone production in women that are younger than 35 compared to those that are older than 40 years. Multivariate analyses were used to adjust for the FF FSH and hCG concentrations and to account for the clustered nature of the data. RESULTS: There is no significant difference in intrafollicular estradiol, between the two age groups, after adjusting for the follicular FSH and hCG levels. The change in follicular estradiol per unit change in FSH is no different in younger versus older women. The estrogen/testosterone ratio and the follicular progesterone levels are also not significantly different in the two age groups after adjusting for follicular FSH and hCG levels. CONCLUSIONS: The somatic cell compartment appears to retain its function in terms of steroidogenic capacity and ability to respond to intrafollicular gonadotropins with aging. Supported by: None.
OBESITY AND METABOLISM P-47 THE INFLUENCE OF BODY MASS INDEX ON OVARIAN STIMULATION AND ICSI OUTCOMES ON WOMEN WITH NORMAL AND OVERWEIGHT. R. C. Ferreira, G. Halpern, C. T. Tanil, F. F. Pasqualotto, A. Iaconelli, Jr, E. Borges, Jr. Sapientiae Institute, Sao Paulo, Brazil; Fertility - Assisted Fertilization Center, Sao Paulo, Brazil; Institute of Biotechnology, University of Caxias do Sul, Caxias do Sul, Brazil. OBJECTIVE: Obesity have been becomes a worldwide epidemic and it serious impact on the various aspects of health. It is known that reproductive potential in obese women is decreased and to be associated with sub-optimal outcomes after assisted reproductive technologies. The aim of the present study was to evaluate the effects of increased body mass index (BMI) on clinical outcomes of ICSI cycles. DESIGN: Case-control study. MATERIALS AND METHODS: It was analyzed 412 ICSI cycles at a private assisted reproduction center. Controlled ovarian stimulation was achieved by a long pituitary down regulation followed by ovarian stimulation with recombinant-FSH and ovum pick up 35 hours after recombinanthCG trigger. The cycles were classified according to BMI: normal-BMI (19.0-24.9 kg/m2, n¼313) and overweight-obesity (R25 kg/m, n¼99). A Pearson correlation was also calculated to establish the association between BMI and maternal age. Regression analysis was done to model the probability of BMI influences on dose of FSH administered, ovarian response (number of MII oocytes retrieved), implantation, pregnancy and miscarriage rates, adjusted to maternal age, in each group. P<0.05 were considered significant. RESULTS: There was a positive correlation between BMI and maternal age (Coef.¼ 0.150; p¼0.001). The linear regression analysis showed that BMI influence the dose of FSH administered on overweight-obesity group (Coef.¼ 42.23; p¼0.043), but not on the normal group (Coef.¼ 18.19; p¼0.442). It was not observed any influence of the BMI on the number of MII recovered (normal-BMI: Coef.¼ -0.284; p¼0.180, overweight-obesity: Coef.¼ 0.129; p¼0.489), implantation rate (normal-BMI: Coef.¼ 1.331; p¼0.365, overweight-obesity: Coef.¼ -0.139; p¼0.871), pregnancy rate (normal-BMI: Coef.¼ -0.038; p¼0.644, overweight-obesity: Coef.¼ -0.062; p¼0.452), or miscarriage rate (normal-BMI: Coef.¼ -0.128; p¼0.615, overweight-obesity: Coef.¼ 0.066; p¼0.782) on both groups. CONCLUSIONS: Our finding showed that overweight and obese patients had a higher dose of FSH administered during controlled ovarian stimulation. Nevertheless, the ovarian response and clinical outcomes were not influenced by the BMI, probably because of higher FSH dose administered compensated the further outcomes. Also, the positive correlation between maternal age and BMI suggest that sub-optimal results on overweight and obese described by other authors, may be actually due to maternal age. Supported by: None.
Vol. 90, Suppl 1, September 2008
S126
Abstracts
MATERIALS AND METHODS: 304 patients were divided into two groups by age: Group I age % 35 years and Group II age R 40. A total of 429 individual follicular fluid (FF) aspirates were collected at the time of oocyte retrieval. Each patient had 1 or 2 individual follicle aspirates collected. Concentrations of FSH, hCG, estradiol, progesterone, and testosterone were determined by automated chemiluminescent immunoassays on the DPC-Immulite 2000 (Diagnostic Products, Los Angeles, CA). Linear regression was initially performed to evaluate the difference in hormone production in women that are younger than 35 compared to those that are older than 40 years. Multivariate analyses were used to adjust for the FF FSH and hCG concentrations and to account for the clustered nature of the data. RESULTS: There is no significant difference in intrafollicular estradiol, between the two age groups, after adjusting for the follicular FSH and hCG levels. The change in follicular estradiol per unit change in FSH is no different in younger versus older women. The estrogen/testosterone ratio and the follicular progesterone levels are also not significantly different in the two age groups after adjusting for follicular FSH and hCG levels. CONCLUSIONS: The somatic cell compartment appears to retain its function in terms of steroidogenic capacity and ability to respond to intrafollicular gonadotropins with aging. Supported by: None.
OBESITY AND METABOLISM P-47 THE INFLUENCE OF BODY MASS INDEX ON OVARIAN STIMULATION AND ICSI OUTCOMES ON WOMEN WITH NORMAL AND OVERWEIGHT. R. C. Ferreira, G. Halpern, C. T. Tanil, F. F. Pasqualotto, A. Iaconelli, Jr, E. Borges, Jr. Sapientiae Institute, Sao Paulo, Brazil; Fertility - Assisted Fertilization Center, Sao Paulo, Brazil; Institute of Biotechnology, University of Caxias do Sul, Caxias do Sul, Brazil. OBJECTIVE: Obesity have been becomes a worldwide epidemic and it serious impact on the various aspects of health. It is known that reproductive potential in obese women is decreased and to be associated with sub-optimal outcomes after assisted reproductive technologies. The aim of the present study was to evaluate the effects of increased body mass index (BMI) on clinical outcomes of ICSI cycles. DESIGN: Case-control study. MATERIALS AND METHODS: It was analyzed 412 ICSI cycles at a private assisted reproduction center. Controlled ovarian stimulation was achieved by a long pituitary down regulation followed by ovarian stimulation with recombinant-FSH and ovum pick up 35 hours after recombinanthCG trigger. The cycles were classified according to BMI: normal-BMI (19.0-24.9 kg/m2, n¼313) and overweight-obesity (R25 kg/m, n¼99). A Pearson correlation was also calculated to establish the association between BMI and maternal age. Regression analysis was done to model the probability of BMI influences on dose of FSH administered, ovarian response (number of MII oocytes retrieved), implantation, pregnancy and miscarriage rates, adjusted to maternal age, in each group. P<0.05 were considered significant. RESULTS: There was a positive correlation between BMI and maternal age (Coef.¼ 0.150; p¼0.001). The linear regression analysis showed that BMI influence the dose of FSH administered on overweight-obesity group (Coef.¼ 42.23; p¼0.043), but not on the normal group (Coef.¼ 18.19; p¼0.442). It was not observed any influence of the BMI on the number of MII recovered (normal-BMI: Coef.¼ -0.284; p¼0.180, overweight-obesity: Coef.¼ 0.129; p¼0.489), implantation rate (normal-BMI: Coef.¼ 1.331; p¼0.365, overweight-obesity: Coef.¼ -0.139; p¼0.871), pregnancy rate (normal-BMI: Coef.¼ -0.038; p¼0.644, overweight-obesity: Coef.¼ -0.062; p¼0.452), or miscarriage rate (normal-BMI: Coef.¼ -0.128; p¼0.615, overweight-obesity: Coef.¼ 0.066; p¼0.782) on both groups. CONCLUSIONS: Our finding showed that overweight and obese patients had a higher dose of FSH administered during controlled ovarian stimulation. Nevertheless, the ovarian response and clinical outcomes were not influenced by the BMI, probably because of higher FSH dose administered compensated the further outcomes. Also, the positive correlation between maternal age and BMI suggest that sub-optimal results on overweight and obese described by other authors, may be actually due to maternal age. Supported by: None.
Vol. 90, Suppl 1, September 2008