RESUMO
Hormonal changes are caused by the menstrual cycle phases, which influence resting metabolic rate and eating behavior. The aim of the study was to determine resting metabolic rate (RMR) and its association with dietary intake according to the menstrual cycle phase in lean and obese Chilean women. This cross-sectional analytical study included 30 adult women (15 lean and 15 with obesity). Body composition was measured with a tetrapolar bioelectrical impedance meter. Nutritional status was determined by adiposity. A 24-h recall of three nonconsecutive days verifies dietary intake. The RMR was measured by indirect calorimetry. All measurements were performed in both the follicular and luteal phases of the menstrual cycle. Statistical analyses were performed with STATA software at a significance level, which was α = 0.05. The RMR (ß = 121.6 kcal/d), temperature (ß = 0.36 °C), calorie intake (ß = 317.1 kcal/d), and intake of lipids (ß = 13.8 g/d) were associated with the luteal phase in lean women. Only extracellular water (ß = 1.11%) and carbohydrate consumption (ß = 45.2 g/d) were associated in women with obesity. Lean women showed increased RMR, caloric intake, and lipid intake during the luteal phase. For women with obesity, carbohydrate intake increased but not RMR.
Assuntos
Metabolismo Basal , Fase Luteal , Adulto , Carboidratos , Estudos Transversais , Metabolismo Energético , Feminino , Humanos , Fase Luteal/metabolismo , Obesidade/metabolismoRESUMO
STUDY QUESTION: What is the role of the endocannabinoid system (eCS) in the alterations of the endocrine system in a murine model of lipopolysaccharide (LPS)-induced miscarriage? SUMMARY ANSWER: In 7-days pregnant wild type, but not cannabinoid receptor type 1 knockout (CB1-KO) mice, LPS increased COX-2 expression and prostaglandin F2α (PGF2α) production in the uterus leading to lower expression of prolactin receptor in the ovary and a marked regression of corpora lutea (CL), suggesting that the eCS mediates the deleterious effects of LPS on reproductive events. WHAT IS KNOWN ALREADY: Appropriate systemic progesterone levels are critical for a successful pregnancy outcome. Precocious loss of luteal progesterone (P4) secretion leads to miscarriage in rodents. We have previously shown that LPS administration to pregnant mice induces embryonic resorption accompanied by a dramatic decrease in systemic progesterone levels in a murine model of inflammatory miscarriage, with the eCS mediating these LPS-induced deleterious effects. STUDY DESIGN SAMPLES/MATERIALS, METHODS: CD1 wild-type (WT) and CB1-KO mice were randomly allocated to Vehicle (saline; i.p.) or LPS (0.5 µg/g body weight; i.p.) treated groups: (WT-Vehicle; WT-LPS; CB1-KO-Vehicle and CB1-KO-LPS). A single injection was given on day 7 of pregnancy and tissues (blood, ovary, uterus) were collected 6, 12, 24 and 48 h later. P4 and PGF2α plasma levels were determined by radioimmunoassay. Cyclooxygenase-2 (COX-2) mRNA (RT-PCR) and protein (Western blot) content in uterus was assayed. COX-2 and prolactin receptor (PrlR) mRNA levels in the ovary were assayed by RT-PCR. Tissue morphology of the CL was assessed by haematoxylin-eosin staining. MAIN RESULTS AND THE ROLE OF CHANCE: Treatment of 7-day pregnant WT mice with LPS induced a P4 withdrawal (p < 0.05), increased in uterine COX-2 mRNA and protein expression (p < 0.05) as well as an increase in uterine PGF2α production (p < 0.05). These changes were absent in LPS-treated 7-day pregnant CB1-KO mice. In ovarian tissues, LPS treatment to 7-day pregnant WT mice induced a downregulation of PrlR mRNA expression (p < 0.05) together with an increase in COX-2 mRNA expression (p < 0.05) and PGF2α content (p < 0.05). These effects were absent in the CB1-KO mice. Collectively, our results suggest a role for the eCS mediating LPS-induced deleterious effects on reproductive tissues. LIMITATIONS, REASONS FOR CAUTION: An important caveat of this study is the endocrine differences between mice and humans during pregnancy (e.g. P4 is produced by the CL throughout pregnancy in mice, whereas this is not the case in humans), which limits the extrapolation of the results presented here. WIDER IMPLICATIONS OF THE FINDINGS: Our findings provide new insights in the role of the endocannabinoid system in the physiopathology of reproduction as well as the role of this endogenous system as a mediator of LPS deleterious effects on reproductive tissues. LARGE SCALE DATA: None. STUDY FUNDING AND COMPETING INTERESTS: Dr Ana María Franchi was funded by Agencia Nacional para la Promoción Científica y Tecnológica (PICT 2010/0813 and PICT 2013/0097) and by Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 2012/0061). The authors have no competing interests.
Assuntos
Aborto Espontâneo/tratamento farmacológico , Aborto Espontâneo/metabolismo , Endocanabinoides/metabolismo , Lipopolissacarídeos/toxicidade , Fase Luteal/metabolismo , Progesterona/metabolismo , Animais , Corpo Lúteo/metabolismo , Feminino , Luteólise/metabolismo , Camundongos , Camundongos Knockout , Gravidez , RadioimunoensaioRESUMO
The aim was to determine the progesterone profile after the introduction of bucks during the advanced luteal phase of does. Fourteen does received vaginal sponges impregnated with 40 mg fluorogestone acetate for 12 days, and a luteolytic dose of a prostaglandin analogue (75 μg of D-cloprostenol) 2 days before sponge removal. Fifteen days after sponge withdrawal one buck was introduced in one of the pens (BE group; n = 6), while the female goats in the other pen remained as controls (CON group; n = 8). The buck was replaced every 24 h, alternating their presence until the end of the experiment. Serum progesterone levels were used to monitor ovarian activity. Progesterone concentration from day 14 to 20 varied with time (P < 0.0001), and there was an interaction between treatment and day (P = 0.02). While progesterone concentration increased from day 15 to day 16 in BE does (P = 0.01), there were no changes in CON does on those days (P = 0.2). On the other hand, progesterone concentrations decreased in BE does from day 18 to day 19 (P = 0.02), without changes in CON does (P = 0.6). Finally, there was a sharp decrease from day 19 to day 20 in both BE (P = 0.0009) and CON (P < 0.0001) does. Overall, our results demonstrated that the introduction of bucks during the late luteal phase of isolated does can induce changes in the progesterone pattern, showing an early increase followed by a pronounced withdrawn.
Assuntos
Feminino , Animais , Cabras/anatomia & histologia , Cabras/fisiologia , Fase Luteal/metabolismo , Progesterona/análise , Ciclo Estral , LuteóliseRESUMO
The aim was to determine the progesterone profile after the introduction of bucks during the advanced luteal phase of does. Fourteen does received vaginal sponges impregnated with 40 mg fluorogestone acetate for 12 days, and a luteolytic dose of a prostaglandin analogue (75 μg of D-cloprostenol) 2 days before sponge removal. Fifteen days after sponge withdrawal one buck was introduced in one of the pens (BE group; n = 6), while the female goats in the other pen remained as controls (CON group; n = 8). The buck was replaced every 24 h, alternating their presence until the end of the experiment. Serum progesterone levels were used to monitor ovarian activity. Progesterone concentration from day 14 to 20 varied with time (P < 0.0001), and there was an interaction between treatment and day (P = 0.02). While progesterone concentration increased from day 15 to day 16 in BE does (P = 0.01), there were no changes in CON does on those days (P = 0.2). On the other hand, progesterone concentrations decreased in BE does from day 18 to day 19 (P = 0.02), without changes in CON does (P = 0.6). Finally, there was a sharp decrease from day 19 to day 20 in both BE (P = 0.0009) and CON (P < 0.0001) does. Overall, our results demonstrated that the introduction of bucks during the late luteal phase of isolated does can induce changes in the progesterone pattern, showing an early increase followed by a pronounced withdrawn.(AU)
Assuntos
Animais , Feminino , Cabras/anatomia & histologia , Cabras/fisiologia , Fase Luteal/metabolismo , Progesterona/análise , Luteólise , Ciclo EstralRESUMO
OBJECTIVE: To quantitate 2-methoxyestradiol (2-ME) in human corpus luteum (CL) of different ages and to determine the expression of cytochrome-P450-1A1 (CYP1A1) and catechol-O-methyl transferase (COMT) in CL and the action of 2-ME on P, vascular endothelial growth factor (VEGF) secretion, and luteal angiogenesis. DESIGN: Experimental study. SETTING: University division of reproductive endocrinology. PATIENT(S): Twenty-four women of reproductive age. INTERVENTION(S): CL was collected from 15 women during the minilaparotomy for tubal sterilization. Granulosa lutein cells were harvested 36 hours after hCG administration in patients undergoing IVF. MAIN OUTCOMES MEASURE(S): Levels of 2-ME were determined by high-performance liquid chromatography in CL. CYP1A1 and COMT were assessed by immunohistochemistry and Western blot. P and VEGF were measured by radioimmunoassay and ELISA. The angiogenic potential was analyzed using EA.hy926 cells. RESULT(S): Plasma levels of E2 decreased in the late luteal phase in association with an increase in luteal tissue of 2-ME concentrations. Concomitantly, there was a significant reduction of angiogenic activity in late CL. There was no significant variation in CYP1A1 and COMT expression in all CL. In physiological doses, 2-ME inhibited basal VEGF by granulosa lutein cells and diminished the angiogenic activity in conditioned media but did not prevent P and VEGF production stimulated by hCG. CONCLUSION(S): These data suggest the participation of 2-ME in physiological luteolysis by reducing angiogenesis. However, 2-ME did not prevent in vitro hCG stimulation of P biosynthesis, providing a mechanism for CL rescue in the cycle of conception.
Assuntos
Corpo Lúteo/irrigação sanguínea , Corpo Lúteo/metabolismo , Estradiol/análogos & derivados , Células Lúteas/metabolismo , Fase Luteal/metabolismo , Neovascularização Fisiológica , Progesterona/biossíntese , 2-Metoxiestradiol , Adulto , Catecol O-Metiltransferase/metabolismo , Linhagem Celular , Meios de Cultivo Condicionados/metabolismo , Citocromo P-450 CYP1A1/metabolismo , Estradiol/sangue , Estradiol/metabolismo , Feminino , Humanos , Fase Luteal/sangue , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
OBJECTIVE: To evaluate nuclear factor kappaB (NF-κB) activation and NF-κB-p65 subunit activation, immunolocalization, and expression in the endometrium of healthy women and endometriosis patients throughout the menstrual cycle. DESIGN: Prospective observational study. SETTING: Affiliated hospital and university research laboratory. PATIENT(S): Twenty-four healthy women and 24 endometriosis patients. INTERVENTION(S): Menstrual, proliferative, and secretory endometrial biopsies. MAIN OUTCOME MEASURE(S): Assessment of NF-κB and p65 activation by protein-DNA binding assays and p65 localization and expression by immunohistochemistry. RESULT(S): Total NF-κB-DNA binding was constitutive and variable in human endometrium accross the menstrual cycle. Healthy women (physiologic conditions) showed higher p65-DNA binding in proliferative than in menstrual and secretory endometrium. Conversely, in endometriosis patients, p65-DNA binding was higher in proliferative and secretory endometrium than in menstrual endometrium. Endometrial epithelial cells showed higher p65 expression level score than endometrial stromal cells. CONCLUSION(S): NF-κB activity is constitutive, physiologic, and variable in human endometrium. The physiologic cyclic p65 activation pattern was altered in endometriosis patients, showing no cyclic variation between the proliferative and secretory phase of the menstrual cycle. The absence of decreased p65 activity in secretory endometrium from endometriosis patients is concurrent with progesterone resistance and could participate in endometrial biologic alterations during the implantation window in endometriosis patients.
Assuntos
Endometriose/metabolismo , Endométrio/metabolismo , Ciclo Menstrual/metabolismo , NF-kappa B/metabolismo , Adulto , Sítios de Ligação , Biópsia , Estudos de Casos e Controles , Chile , DNA/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Endometriose/patologia , Endométrio/patologia , Células Epiteliais/metabolismo , Feminino , Fase Folicular/metabolismo , Humanos , Imuno-Histoquímica , Fase Luteal/metabolismo , Menstruação/metabolismo , Pessoa de Meia-Idade , Estudos Prospectivos , Células Estromais/metabolismo , Fator de Transcrição RelA/metabolismo , Adulto JovemRESUMO
OBJECTIVE: To investigate the participation of catecholamines in the association between peripheral innervation and luteal steroidogenesis. DESIGN: Animal study. SETTING: University animal laboratory. ANIMAL(S): Six to eight virgin adult Holtzman-strain female rats in control and experimental groups on diestrus days 1 and 2. INTERVENTION(S): Removal of the coeliac ganglion-superior ovarian nerve-ovary system, with catecholaminergic agonist or antagonist added in the ganglion compartment (experimental group only). The control group received no treatment. MAIN OUTCOME MEASURE(S): Ovarian neurotransmitters and their catabolites measured by reverse-phase high-pressure liquid chromatography, and A(2) measured by radioimmunoassay. RESULT(S): On day 1, dopamine and catabolite increased whereas norepinephrine decreased, and the noradrenergic neuronal activity index was higher. On day 2, dopamine levels decreased, norepinephrine increased, and dopaminergic neuronal activity was higher. The release of A(2) was decreased by addition of norepinephrine to the ganglions on day 1, but was increased by the norepinephrine antagonist on day 2. Hence, norepinephrine increased A(2) release, and propranolol diminished it. CONCLUSION(S): Ganglionic activity is modified by noradrenergic stimulus, leading to different ovarian A(2) release profiles. The peripheral nervous system is a modulator in these homeostatic mechanisms.
Assuntos
Androstenodiona/metabolismo , Plexo Celíaco/metabolismo , Fase Luteal/metabolismo , Norepinefrina/metabolismo , Ovário/metabolismo , Adrenérgicos/farmacologia , Animais , Plexo Celíaco/efeitos dos fármacos , Feminino , Fase Luteal/efeitos dos fármacos , Ovário/efeitos dos fármacos , Ratos , Ratos Sprague-DawleyRESUMO
To investigate the luteal phase endometrial expression of leukemia inhibitor factor (LIF), insulin-like growth factor 1 (IGF-1), progesterone receptor (PR), claudin 4 (CLDN4), vascular-endothelial growth factor receptor 3 (VEGFR-3), bone morphogenetic protein 4 (BMP-4) and citokeratin 7 (CK-7), we obtained luteal phase endometrial samples from 52 women. Samples were dated and integrated using a tissue microarray (TMA). Samples were immunostained for LIF, IGF-1, PR, CLDN4, VEGFR-3, BMP-4 and CK-7. Frequencies of positive expressions at the early, mid and late luteal phases were compared by two proportions test. Concomitant expression of these proteins was assessed with Chi-square or Fischer's test. The frequency of LIF was positively correlated to the frequency of IGF-1 (r = 0.99; p < 0.05) and PR (r = 0.99; p < 0.05), and the correlation between IGF-1 and PR tended to be significant (r = 0.98; p < 0.1). The expression of PR was associated with the absence of CLDN4 (p < 0.001). Thus, expression of LIF, IGF-1 and PR are correlated during the luteal phase, and immunohistochemistry for these proteins might be used to assist in the assessment of endometrial maturation. In addition, the expression of CLDN4 and PR was not concomitant, warranting further investigation on the relationship of their endometrial expression.
Assuntos
Endométrio/metabolismo , Fase Luteal/metabolismo , Adulto , Proteína Morfogenética Óssea 4/metabolismo , Distribuição de Qui-Quadrado , Claudina-4 , Feminino , Humanos , Imuno-Histoquímica , Fator de Crescimento Insulin-Like I/metabolismo , Queratina-7/metabolismo , Fator Inibidor de Leucemia/metabolismo , Proteínas de Membrana/metabolismo , Seleção de Pacientes , Análise Serial de Proteínas , Receptores de Progesterona/metabolismo , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/metabolismoRESUMO
BACKGROUND: Claudin-4 (CLDN4) is one of several proteins that act as molecular mediators of embryo implantation. Recently, we examined immunolabeling of leukemia inhibitory factor (LIF) in the endometrial tissue of 52 IVF patients, and found that LIF staining intensity was strongly correlated with successful pregnancy initiation. In the same set of patients, we have now examined endometrial CLDN4 expression, to see how expression intensity may vary with LIF. We examined CLDN4 in the luteal phase of the menstrual cycle, immediately preceding IVF treatment. Our aim was to compare expression of LIF and CLDN4 in the luteal phase, and document these patterns as putative biomarkers for pregnancy. METHODS: Endometrial tissue was collected from women undergoing IVF. Endometrial biopsies were obtained during the luteal phase preceding IVF, and were then used for tissue microarray (TMA) immunolabeling of CLDN4. Previously published LIF expression data were then combined with CLDN4 expression data, to determine CLDN4/LIF expression patterns. Associations between successful pregnancy after IVF and combined CLDN4/LIF expression patterns were evaluated. RESULTS: Four patterns of immunolabeling were observed in the endometrial samples: 16% showed weak CLDN4 and strong LIF (CLDN4-/LIF+); 20% showed strong CLDN4 and strong LIF (LIF+/CLDN4+); 28% showed strong CLDN4 and weak LIF (CLDN4+/LIF-); and 36% showed weak CLDN4 and weak LIF (CLDN4-/LIF-). Successful implantation after IVF was associated with CLDN4-/LIF+(p = 0.003). Patients showing this endometrial CLDN4-/LIF+ immunolabeling were also 6 times more likely to achieve pregnancy than patients with endometrial CLDN4+/LIF- immunolabeling (p = 0.007). CONCLUSION: The combined immunolabeling expression of CLDN4-/LIF+ in endometrial tissue is a potential biomarker for predicting successful pregnancy in IVF candidates.
Assuntos
Endométrio/metabolismo , Fertilização in vitro , Fator Inibidor de Leucemia/metabolismo , Proteínas de Membrana/metabolismo , Adulto , Biomarcadores/metabolismo , Coeficiente de Natalidade , Claudina-4 , Implantação do Embrião/fisiologia , Feminino , Humanos , Imuno-Histoquímica , Infertilidade/metabolismo , Fator Inibidor de Leucemia/genética , Fator Inibidor de Leucemia/fisiologia , Fase Luteal/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/fisiologia , Gravidez , Resultado do TratamentoRESUMO
OBJECTIVES: Fibroadenoma is the most common benign mammary condition among women aged 35 or younger. Expression of Ki-67 antigen has been used to compare proliferative activity of mammary fibroadenoma epithelium in the follicular and luteal phases of the menstrual cycle. MATERIALS AND METHODS: Ninety eumenorrheic women were selected for tumour excision; they were assigned to either of the two groups, according to their phase of menstrual cycle. At the end of the study, 75 patients with 87 masses were evaluated by epithelial cell Ki-67 expression, blind (no information given concerning group to which any lesion belonged). RESULTS: Both groups were found to be homogeneous relative to age, menarche, body mass index, previous gestation, parity, breastfeeding, number of fibroadenomas, family history of breast cancer and tabagism. Median tumour size was 2.0 cm and no relationship between proliferative activity and nodule diameter was observed. No typical pattern was observed in the expression of Ki-67 in distinct nodules of the same patient. Average values for expression of Ki-67 (per 1000 epithelial cells) in follicular and luteal phases were 27.88 and 37.88, respectively (P = 0.116). CONCLUSION: Our findings revealed that proliferative activities in the mammary fibroadenoma epithelium did not present a statistically significant difference in the follicular and luteal phases. The present study contributes to clarifying that fibroadenoma is a neoplasm and does not undergo any change in the proliferative activity during the menstrual cycle.