RESUMO
Understanding the regulatory mechanisms that affect obesogenic genes expression in newborns is essential for early prevention efforts, but they remain unclear. Our study aimed to explore whether the maternal p-BMI and GWG were associated with regulatory single-locus DNA methylation in selected obesogenic genes. For this purpose, DNA methylation was assayed by Methylation-Sensitive High Resolution Melting (MS-HRM) technique and Sanger allele-bisulfite sequencing in fifty samples of umbilical vein to evaluate glucosamine-6-phosphate deaminase 2 (GNPDA2), peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC1α), and leptin receptor (LEPR) genes. Correlations between DNA methylation levels and indicators of maternal nutritional status were carried out. Western blotting was used to evaluate protein expression in extracts of the same samples. Results indicated that GNPDA2 and PGC1α genes have the same level of DNA methylation in all samples; however, a differential DNA methylation of LEPR gene promoter was found, correlating it with GWG and this correlation is unaffected by maternal age or unhealthy habits. Furthermore, leptin receptor (Lep-Rb) was upregulated in samples that showed the lowest levels of DNA methylation. This study highlights the association between poor GWG and adjustments on obesogenic genes expression in newborn tissues with potential consequences for development of obesity in the future.
Assuntos
Aldose-Cetose Isomerases/metabolismo , Metilação de DNA , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Receptores para Leptina/metabolismo , Veias Umbilicais/metabolismo , Adolescente , Adulto , Aldose-Cetose Isomerases/genética , Índice de Massa Corporal , Feminino , Ganho de Peso na Gestação , Humanos , Estado Nutricional , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/genética , Gravidez , Primeiro Trimestre da Gravidez , Regiões Promotoras Genéticas , Receptores para Leptina/genética , Adulto JovemRESUMO
DNA methylation status of RXRα gene promoter has been correlated with maternal diet during early pregnancy, and associated with offspring's adiposity and bone mineral content. In adult life, increased methylation of RXRα promoter region is associated with myocardium pathologies. Early growth response proteins (EGR) are zinc finger transcription factors associated with several cellular pathways such as inflammation, apoptosis, and cardiopathies. DNA-binding sequences of EGR proteins have been reported in the RXRα gene promoter using chromatin immunoprecipitation methods. Here, we used correlations between the maternal pre-pregnancy body mass index (p-BMI), gestational weight gain (GWG), and birth weight (BW) as indirect indicators of the maternal nutritional status as modifier of DNA methylation in the offspring. DNA methylation status from newborns' umbilical vein blood in full-term pregnancy was evaluated in a short sequence (116 pb) of the RXRα gene promoter that contains the elements of response sequence for EGR proteins. Fifty-three bisulfite-modified DNA samples were assessed through methyl-sensitive high-resolution melting (MS-HRM) analysis. To validate the results, we directly sequenced MS-HRM samples to confirm the presence of CpG-methylated positions. In addition, the RXRα protein levels in extracts of umbilical vein blood were evaluated by western blot. We found differential methylation in a specific locus of the RXRα promoter surrounding the EGR-binding sequence; however, no correlation was found with the level of RXRα protein expression. Variability in the methylation status of the RXRα promoter near the EGR transcription factor binding site in newborn cord blood provides controversial epigenetic insights into RXRα regulation via EGR proteins.
Assuntos
Metilação de DNA , Sangue Fetal , Loci Gênicos , Elementos de Resposta , Receptor X Retinoide alfa/genética , Adolescente , Adulto , Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Receptor X Retinoide alfa/metabolismoRESUMO
BACKGROUND: The activity of matrix degrading enzymes plays a leading role in the rupture of the fetal membranes under normal and pathological human labor, and matrix metalloproteinase-9 (MMP-9) it is considered a biomarker of this event. To gain further insight into local MMP-9 origin and activation, in this study we analyzed the contribution of human placental leukocytes to MMP-9 secretion and explored the local mechanisms of the pro-enzyme activation. METHODS: Placental blood leukocytes were obtained from women at term gestation without labor and maintained in culture up to 72 h. MMP-9 activity in the culture supernatants was determined by zymography and using a specific substrate. The presence of a potential pro-MMP-9 activator in the culture supernatants was monitored using a recombinant biotin-labeled human pro-MMP-9. To characterize the endogenous pro-MMP-9 activator, MMP-1, -3, -7 and -9 were measured by multiplex assay in the supernatants, and an inhibition assay of MMP-9 activation was performed using an anti-human MMP-3 and a specific MMP-3 inhibitor. Finally, production of MMP-9 and MMP-3 in placental leukocytes obtained from term pregnancies with and without labor was assessed by immunofluorescence. RESULTS: Placental leukocytes spontaneously secreted pro-MMP-9 after 24 h of culture, increasing significantly at 48 h (P≤0.05), when the active form of MMP-9 was detected. Culture supernatants activated the recombinant pro-MMP-9 showing that placental leukocytes secrete the activator. A significant increase in MMP-3 secretion by placental leukocytes was observed since 48 h in culture (P≤0.05) and up to 72 h (P≤0.001), when concentration reached its maximum value. Specific activity of MMP-9 decreased significantly (P≤0.005) when an anti-MMP-3 antibody or a specific MMP-3 inhibitor were added to the culture media. Placental leukocytes from term labor produced more MMP-9 and MMP-3 compared to term non-labor cells. CONCLUSIONS: In this work we confirm that placental leukocytes from human term pregnancies are able to secrete large amounts of MMP-9, and that the production of the enzyme it is enhanced by labor. We also demonstrate for the first time that endogenous MMP-3 plays a major role in MMP-9 activation process. These findings support the contribution of placental leukocytes to create the collagenolytic microenvironment that induces the rupture of the fetal membranes during human labor.
Assuntos
Trabalho de Parto/sangue , Trabalho de Parto/metabolismo , Leucócitos/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Placenta/citologia , Sobrevivência Celular , Colágeno/metabolismo , Ativação Enzimática , Precursores Enzimáticos/metabolismo , Feminino , Humanos , Leucócitos/citologia , Leucócitos/enzimologia , GravidezRESUMO
Preterm birth is a public health issue of global significance, which may result in mortality during the perinatal period or may lead to major health and financial consequences due to lifelong impacts. Even though several risk factors for preterm birth have been identified, prevention efforts have failed to halt the increasing rates of preterm birth. Epidemiological studies have identified air pollution as an emerging potential risk factor for preterm birth. However, many studies were limited by study design and inadequate exposure assessment. Due to the ubiquitous nature of ambient air pollution and the potential public health significance of any role in causing preterm birth, a novel focus investigating possible causal mechanisms influenced by air pollution is therefore a global health priority. We hypothesize that air pollution may act together with other biological factors to induce systemic inflammation and influence the duration of pregnancy. Evaluation and testing of this hypothesis is currently being conducted in a prospective cohort study in Mexico City and will provide an understanding of the pathways that mediate the effects of air pollution on preterm birth. The important public health implication is that crucial steps in this mechanistic pathway can potentially be acted on early in pregnancy to reduce the risk of preterm birth.
Assuntos
Poluentes Atmosféricos/análise , Poluição do Ar/efeitos adversos , Exposição Materna/efeitos adversos , Complicações na Gravidez/diagnóstico , Citocinas/metabolismo , Dieta , Feminino , Humanos , Inflamação/complicações , México , Modelos Teóricos , Obesidade/complicações , Trabalho de Parto Prematuro/etiologia , Estresse Oxidativo , Material Particulado , Gravidez , Nascimento Prematuro , Fatores de RiscoRESUMO
PROBLEM: Human parturition is associated with an intrauterine pro-inflammatory environment in the choriodecidua. Evidence that some mediators of this signaling cascade also elicit responses leading to labor prompted us to characterize the cellular sources of these mediators in the human choriodecidua. METHOD OF STUDY: Leukocyte-enriched preparations from human choriodecidua (ChL) and intervillous placental blood leukocytes (PL) were maintained in culture. Secretions of inflammatory cytokines, chemokines, and MMP-9 were documented. Leukocyte phenotype of ChL and PL was determined by flow cytometry using specific fluorochrome-conjugated antibodies. RESULTS AND CONCLUSIONS: ChL showed a distinct pro-inflammatory secretion pattern of cytokines and chemokines when compared with PL, including higher amounts of TNF-α and IL-6, and decreased secretions of IL-4 and IL-1ra. ChL also secreted more MIP-1α and MCP-1 and MMP-9 than PL. No significant differences were found in leukocytes subsets between compartments. Based on our findings, we propose that ChL isolated from fetal membranes at term are functionally different from PL and may collaborate to modulate the microenvironment linked to induction and progression of human labor.
Assuntos
Córion/imunologia , Decídua/imunologia , Leucócitos Mononucleares/imunologia , Subpopulações de Linfócitos/imunologia , Placenta/imunologia , Células Cultivadas , Microambiente Celular , Citocinas/metabolismo , Feminino , Humanos , Imunofenotipagem , Mediadores da Inflamação/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Gravidez , Nascimento a Termo/imunologiaRESUMO
BACKGROUND: Twin pregnancies (TP) have shown a greater risk of adverse perinatal outcomes than singletons have. However, there is still no consensus about whether the TP achieved by assisted reproduction techniques (ART) have worse perinatal outcomes compared with TP achieved spontaneously. OBJECTIVE: Compare the incidence of adverse perinatal outcomes (preterm birth, premature rupture of membranes, gestational diabetes, pregnancy-induced hypertension, anemia, weight and destination of the newborn) in Mexican women with TP achieved spontaneously vs those with TP achieved by ART. METHODS: Historical cohort study with two sample groups: group 1, women with TP achieved spontaneously, and group 2, women with TP achieved by ART. Women with TP achieved by TRA were matched 1:1 with women with EG achieved spontaneously for age, weeks of gestation, chorionicity and body mass index at admission to prenatal care. Adverse perinatal outcomes were compared between the two groups and the odds ratio (OR) had a 95% confidence interval. RESULTS: There were 57 women per group. Baseline characteristics were similar in both groups except for nulliparity (38.6% in group 1 vs 82.5% in group 2 (p < 0.0001)). In group 2 there was a higher incidence of pregnancy-induced hypertension (group 1 (19.3%) vs. group 2 (42.1%), OR 2.5 (95% CI 1.07-5.8)). Newborns in group 2 were admitted to nursery more often than those in group 1 (49.1% vs. 35.1% OR 1.7 IC 95% 1.04-3.04). There was no difference in other adverse perinatal outcomes. CONCLUSIONS: Mexican women with TP achieved by ART had higher risk of pregnancy-induced hypertension compared to women with TP achieved spontaneously.
Assuntos
Gravidez de Gêmeos , Técnicas de Reprodução Assistida/efeitos adversos , Adulto , Feminino , Humanos , México , Gravidez , Resultado da GravidezRESUMO
In the present pilot study, we evaluated the effect of maternal adiposity on the plasma concentration of adipocytokines in pregnant women and their newborns. Twenty patients with term gestations without labour were initially selected by pregestational BMI and then classified into two study groups (n 10 each), according to their median value of adiposity (total body fat). Concentrations of TNF-α, IL-1ß, IL-6, leptin and adiponectin in plasma of maternal peripheral blood and fetal cord blood were measured and correlated to maternal adiposity. Maternal adiposity showed a significant negative correlation with fetal adiponectin (r - 0·587, P = 0·01) and IL-6 (r - 0·466, P = 0·05), a significant positive correlation with maternal leptin (r 0·527, P = 0·02) and no correlation with TNF-α or IL-1ß. Adiponectin was higher in fetal plasma than in maternal plasma (P = 0·043), but significantly lower in newborns from women with high adiposity than in newborns from women with low adiposity (P = 0·040). Our results suggest that fetuses from obese women may be less able to control inflammation, due to lower circulating anti-inflammatory adipocytokines, which could limit their optimal development or even increase the risk of abortion or preterm labour.
Assuntos
Adipocinas/sangue , Adiposidade/fisiologia , Citocinas/sangue , Inflamação/sangue , Adiponectina/sangue , Adulto , Biomarcadores/sangue , Peso ao Nascer , Feminino , Sangue Fetal/química , Idade Gestacional , Humanos , Recém-Nascido , Leptina/sangue , Masculino , Idade Materna , México , Projetos Piloto , Gravidez , Adulto JovemRESUMO
Rupture of the fetal membranes during human labor is associated with an inflammatory process localized to the maternal-fetal interface. There is evidence that specific leukocytes subsets are attracted to the choriodecidua, and that after homing they condition a local inflammatory microenvironment, possibly being directly involved in rupture of the membranes. In this study our aim was to compare the phenotypes and function of leukocytes located in the placental intervillous blood with peripheral leukocytes obtained before or after labor, including expression of modulators of inflammation in these cells. Flow cytometry revealed that the proportion of CD14(+) cells is increased in intervillous blood, suggesting the participation of monocytes/macrophages during labor. Real time qRT-PCR showed that at term gestation and particularly during labor, placental blood leukocytes adopt a different expression pattern of pro-inflammatory cytokines than leukocytes in peripheral blood, including IL-1beta and IL-1RA. During labor, both placental and peripheral leukocytes increase their secretion of matrix metalloproteinase-9. Moreover, we showed that placental leukocytes respond differently than peripheral leukocytes to bacterial lipopolysaccharide, secreting differential amounts of TNF-alpha, IL-1beta and IL-6. Finally, a preliminary proteomic characterization of placental leukocytes revealed a significantly higher number of individual proteins than in peripheral leukocytes. Our results support the existence of selective subsets of leukocytes recruited to the maternal-fetal interface that may participate in the triggering of parturition.
Assuntos
Trabalho de Parto/imunologia , Leucócitos/imunologia , Placenta/imunologia , Membranas Extraembrionárias/imunologia , Feminino , Humanos , Inflamação/imunologia , Proteína Antagonista do Receptor de Interleucina 1/imunologia , Interleucina-1beta/imunologia , Interleucina-6/imunologia , Receptores de Lipopolissacarídeos/imunologia , Macrófagos/imunologia , Metaloproteinase 9 da Matriz/imunologia , Monócitos/imunologia , Polissacarídeos Bacterianos/imunologia , Gravidez , Fator de Necrose Tumoral alfa/imunologiaRESUMO
BACKGROUND: The human labor is an inflammatory process invading leukocytes modulated by gestational tissues. The local increase of cell adhesion molecules (CAMs) promotes the permanence of these leukocytes in the coriodecidua. Gestational tissues express ICAM-1, while circulating leukocytes expressing its ligand Mac-1. OBJECTIVE: To analyze, first, if the expression of CAMs in the fetal membranes is associated with progress of gestational age, and second, the expression of CAMs on circulating leukocytes in the uterus (placenta). MATERIAL AND METHOD: original and closed study conducted at the Instituto Nacional de Perinatologia Isidro Espinosa de los Reyes (Mexico City). We included samples from healthy women between 15 and 44 years of age with term pregnancies (> or =37 weeks gestation). RESULTS: Real time PCR analysis showed that the expression of CAMs in the fetal membranes remained constant before labor. ICAM3 and ICAM1 tended to increase during labor, while ICAM2, VCAM1, SELE and SELP decrease with advancing gestational age. Placental leukocytes showed a clear increase in the expression of ITGAM (Mac-1) during labor. CONCLUSIONS: These results show that the maternal-fetal interface expresses a specific combination of CAMs during labor, including ICAM1, ICAM-3 and Mac-1. The expression of these molecules could promote the retention of leukocytes in the local tissues to modulate the local inflammatory microenvironment during human labor.
Assuntos
Moléculas de Adesão Celular/metabolismo , Membranas Extraembrionárias/metabolismo , Trabalho de Parto/metabolismo , Leucócitos/metabolismo , Terceiro Trimestre da Gravidez/metabolismo , Adolescente , Adulto , Distinções e Prêmios , Células Sanguíneas/metabolismo , Quimiotaxia de Leucócito , Sistemas Computacionais , Feminino , Idade Gestacional , Ginecologia , Humanos , Inflamação , México , Obstetrícia , Placenta/citologia , Placenta/metabolismo , Reação em Cadeia da Polimerase , Gravidez , Adulto JovemRESUMO
OBJECTIVE: To evaluate the secretion of interleukin 6 (IL-6) in cervicovaginal fluid in a pseudocohort that emulates the evolution of the labor. MATERIAL AND METHOD: Samples of cervicovaginal fluid of patients with 20 to 40 weeks of gestation were taken, patients were classified in 5 strata of the development of the labor. Each stratum reflects the progressive activation of the uterine activity and the cervical changes. To each sample was made determination of IL-6 by means of the system Multiplex. The samples of patient with infection data were eliminated. RESULTS: 173 samples were included distributed in each one of the five strata. Basal secretion exists of IL-6 to the cervicovaginal fluid along the gestation, that doesn't modify until the active labor appears. Only when the uterine activity is manifested in form to regulate and effective, very significant increase is documented in the concentration of the IL-6 in the cervicovaginal fluid. CONCLUSIONS: The IL-6 it is a proinflammatory cytokine that increases in a specific way in the moment in that the effective uterine activity begins, thus, it is an excellent candidate to be evaluated as marker presage of the event of normal labor and preterm.