ABSTRACT
BACKGROUND: Hyperandrogenic conditions have been associated with a high prevalence of endometrial pathologies related to cell survival. However, the action of androgens on proliferation and apoptosis in endometrial cells is poorly understood. Therefore, the aim of the present study was to evaluate the effect of androstenedione on cell proliferation, cell death and expression of estrogen receptor (ER) isoforms and proteins related to apoptosis in endometrial cells using two in vitro experimental approaches. METHODS: The endometrial tissue was obtained from 20 eumenorrheic women [28.7 (25 - 35) years] during the early secretory phase. We analyzed cell proliferation (immunohistochemistry of Ki-67 and spectrophotometric assay); apoptosis (DNA fragmentation (TUNEL) and Annexin V-FITC binding); ER-alpha, ER-beta bcl-2 and bax mRNA abundance (RT-PCR) in explants and isolated endometrial epithelial (EEC) and stromal cells (ESC) incubated with androstenedione 1 micro mol/l (A4) or A4 plus hydroxyflutamide 10 micro mol/l (F) for 24 h. RESULTS: In explants, A4 induced an increase of cell proliferation and a decrease on apoptosis in the stromal compartment (p < 0.05). In isolated ESC, proliferation augmented with A4 (p < 0.05), whereas, no significant modifications in the expression of ER-alpha, ER-beta bcl-2 and bax nor in the apoptotic index were observed. In EEC, A4 increase the ER-beta mRNA abundance (p < 0.05) and a decrease of the bcl-2/bax ratio (p < 0.05), without an increase in the apoptotic index. Hydroxyflutamide reverted the effect of androstenedione on the parameters described. CONCLUSIONS: These results indicate that androstenedione may modulate cell survival, expression of ER-beta and proteins related to apoptosis, suggesting a potential mechanism that associates the effect of hyperandrogenemia on the endometrial tissue.
Subject(s)
Androstenedione/pharmacology , Apoptosis/drug effects , Cell Proliferation/drug effects , Endometrium/cytology , Endometrium/drug effects , Stromal Cells/drug effects , Actins/biosynthesis , Adult , Cells, Cultured , Estrogen Receptor alpha/biosynthesis , Estrogen Receptor beta/biosynthesis , Female , Flutamide/analogs & derivatives , Flutamide/pharmacology , Humans , In Situ Nick-End Labeling , Proto-Oncogene Proteins c-bcl-2/biosynthesis , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Stromal Cells/cytology , bcl-2-Associated X Protein/biosynthesisABSTRACT
OBJECTIVE: To investigate the role of nitric oxide (NO) and death regulatory genes, bcl-2 and bax, in human endometria apoptosis. DESIGN: Expression of bcl-2, bax, NO synthases (NOS), and the apoptotic effect of L-arginine on endometrial explants in vitro. SETTING: Prospective study. PATIENT(S): Thirty-seven eumenorrheic women. INTERVENTION(S): Endometrial samples were obtained with Pipelle suction curette after women signed institutional informed consent forms. MAIN OUTCOME MEASURE(S): DNA fragmentation (TUNEL), immunohistochemistry, and reverse transcription polymerase chain reaction. RESULT(S): Apoptosis was detected in mid and late secretory endometria. L-arginine induced an increase in apoptosis in stroma (threefold), glands (eightfold), and surface epithelia (fourfold) in proliferative but not secretory endometria explants. Immunostaining of Bcl-2 was almost absent in the secretory endometria, whereas Bax increased in the stroma at the end of the menstrual cycle, coincident to the decrease in the bcl-2/bax mRNA relative ratio (P<.05) observed in secretory endometria. CONCLUSION(S): The induction of DNA fragmentation by L-arginine on proliferative endometria suggests that NO may be involved in the endometrial apoptotic process, whose control may be related predominantly to the changes of Bcl-2 expression.
Subject(s)
Apoptosis/physiology , Endometrium/physiology , Genes, bcl-2 , Nitric Oxide/physiology , Proto-Oncogene Proteins c-bcl-2 , Proto-Oncogene Proteins/physiology , Adult , Analysis of Variance , Apoptosis/genetics , Endometrium/cytology , Female , Humans , Immunohistochemistry , In Situ Nick-End Labeling , Multigene Family , Organ Culture Techniques , Reference Values , Reverse Transcriptase Polymerase Chain Reaction , bcl-2-Associated X ProteinABSTRACT
Presentamos nuestra experiencia en inducción de ovulación y fertilidad en pacientes con amenorrea hipotalámica (AH) tratadas con Hormona liberadora de gonadotrofina (GNRH) en infusión pulsátil (GnRHp). Se trataron 13 pacientes ciomifeno negativo, de las cuales 10 tenían una AH primaria y 3 una AH secundaria. Ocho del total de las pacientes estaban interesadas en fertilidad. El tratamiento se realizó en forma ambulatorio con bomba de infusión pulsátil (ZykiomatO Alemania) programada para administrar 5 Wg de GNRH cada 90 minutos por vía endovenosa. El desarrollo folicular fue monitorizado mediante ultrasonografía y determinaciones hormonales seriadas. Se indujeron un total de 19 ciclos, 14 fueron ovulatorios con folículo único (73,7 por ciento). Trece inducciones se llevaron a cabo en las 8 pacientes expuestas a embarazo, 7 de los cuales fueron concepcionales (tasa de fecundidad: 61,5 por ciento, tasa de embarazo: 85,5 por ciento). Sólo una paciente abortó espontáneamente. No se observaron complicaciones mayores. En conclusión, la inducción de ovulación con GNRH demostró ser un método de inducción de ovulación efectivo y seguro en pacientes con AH ciomifeno resistentes. Debido a que se conservan los mecanismos de retrocontrol entre ovario e hipótesis, los embarazos múltiples y el síndrome de hiperestimulación ovárica son infrecuentes, lo que permite una monitorización menos estricta de la paciente