RESUMEN
BACKGROUND: Trypanosoma cruzi crosses the placental barrier and produces the congenital transmission of Chagas disease (CD). Structural alterations of the chorionic villi by this parasite have been described in vitro, but little is known about trophoblast turnover in placentas from women with CD. OBJECTIVE: To analyze the proliferation and fusion processes in placentas from women with CD. METHODS: Archived human term placenta paraffin-embedded blocks were used, from women with CD (CDP), and no pathology (NP). Immunohistochemistry tests were performed for Ki67 to calculate the proliferation index (PI) of cytotrophoblast (CTB) and Syncytin-1, a fusion marker of syncytiotrophoblast (STB). Hematoxylin/Eosin stained sections were employed to analyze STB percentages, STB detachment areas and syncytial knots quantity. Non parametric Student's t-tests were performed (p < 0.05). RESULTS: Syncytial knots and STB detachment significantly increased in placental villi from the CDP group. STB percentage was significantly lower in the CDP group as well as the PI and Syncytin-1 expression significantly decreased in these placentas, compared with control (NP). CONCLUSION: Dynamic of trophoblast turnover is altered in placentas from women with CD. These changes may lead into a gap in the placental barrier possibly allowing the parasite entry into the chorionic villi.
Asunto(s)
Enfermedad de Chagas , Trypanosoma cruzi , Femenino , Humanos , Embarazo , Vellosidades Coriónicas/metabolismo , Vellosidades Coriónicas/parasitología , Vellosidades Coriónicas/patología , PlacentaRESUMEN
Congenital toxoplasmosis is represented by the transplacental passage of Toxoplasma gondii from the mother to the fetus. Our studies demonstrated that T. gondii developed mechanisms to evade of the host immune response, such as cyclooxygenase (COX)-2 and prostaglandin E2 (PGE2) induction, and these mediators can be produced/stored in lipid droplets (LDs). The aim of this study was to evaluate the role of COX-2 and LDs during T. gondii infection in human trophoblast cells and villous explants. Our data demonstrated that COX-2 inhibitors decreased T. gondii replication in trophoblast cells and villous. In BeWo cells, the COX-2 inhibitors induced an increase of pro-inflammatory cytokines (IL-6 and MIF), and a decrease in anti-inflammatory cytokines (IL-4 and IL-10). In HTR-8/SVneo cells, the COX-2 inhibitors induced an increase of IL-6 and nitrite and decreased IL-4 and TGF-ß1. In villous explants, the COX-2 inhibitors increased MIF and decreased TNF-α and IL-10. Furthermore, T. gondii induced an increase in LDs in BeWo and HTR-8/SVneo, but COX-2 inhibitors reduced LDs in both cells type. We highlighted that COX-2 is a key factor to T. gondii proliferation in human trophoblast cells, since its inhibition induced a pro-inflammatory response capable of controlling parasitism and leading to a decrease in the availability of LDs, which are essentials for parasite growth.
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Vellosidades Coriónicas/parasitología , Ciclooxigenasa 2/metabolismo , Gotas Lipídicas/metabolismo , Toxoplasma/crecimiento & desarrollo , Trofoblastos/parasitología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Vellosidades Coriónicas/inmunología , Vellosidades Coriónicas/metabolismo , Inhibidores de la Ciclooxigenasa 2/farmacología , Proteínas de la Matriz Extracelular/metabolismo , Interacciones Huésped-Parásitos , Humanos , Interleucinas/metabolismo , Factores Inhibidores de la Migración de Macrófagos/metabolismo , Nitritos/metabolismo , Toxoplasma/inmunología , Factor de Crecimiento Transformador beta/metabolismo , Trofoblastos/inmunología , Trofoblastos/metabolismoRESUMEN
New evidence suggests that glycan expression in placental cells of women with invasive disorders of pregnancy differs from that in normal pregnant women. Hypothesizing that modifications of glycan expression could account for the course of preeclampsia, we established placental villous histocultures and compared glycan expression in women with preeclampsia with that in normal pregnant women and also in syncytialized BeWo cells, and we tested the effect of glycan expression on the functional phenotypes of circulating natural killer (NK) cells. Histocultures of third-trimester placentae from women with preeclampsia and full-term placentae from healthy pregnant women and BeWo choriocarcinoma cells were assessed for the expression of terminal glycans by lectin-binding assays. Circulating NK cells from nonpregnant healthy donors were tested in vitro for their cytotoxic activity and intracellular cytokine content. Histocultures from women with preeclampsia expressed significantly more mannose than did those from healthy pregnant women. Both histocultures and BeWo cells expressed terminal fucose, mannose, sialic acid, and N -acetylgalactosamine, although mean fluorescence intensity (MFI) expression was lower in choriocarcinoma cells than in cells from histocultures. Cocultures of circulating NK cells with K562 target cells resulted in a dose-dependent cytotoxicity effect, but the use of BeWo cells as target reduced cytotoxic activity; this reduction was not affected by syncytialization. Histocultures of placental villous tissue of women with preeclampsia expressed high levels of terminal mannose. We proposethat placental glycans may modulate the functional activity of circulating NK cells in the context of systemic inflammatory response in preeclampsia.
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Vellosidades Coriónicas/patología , Células Asesinas Naturales/inmunología , Polisacáridos/metabolismo , Preeclampsia/inmunología , Adolescente , Adulto , Estudios de Casos y Controles , Comunicación Celular/inmunología , Línea Celular , Vellosidades Coriónicas/inmunología , Vellosidades Coriónicas/metabolismo , Femenino , Glicosilación , Voluntarios Sanos , Humanos , Células Asesinas Naturales/metabolismo , Preeclampsia/sangre , Preeclampsia/patología , Embarazo , Tercer Trimestre del Embarazo , Estudios Prospectivos , Trofoblastos/inmunología , Trofoblastos/metabolismo , Adulto JovenRESUMEN
The use of immunosuppressive drugs guarantees the vitality of the graft and allows gestation in spite of intercurrences such as prematurity and intrauterine growth restriction. However, little is known about the direct effects of immunosuppressive drugs on placental cells. We investigated the effects of immunosuppressive drugs in the chorionic villous explants from human term placentas of healthy gestations. Human placental explants from term gestations (37-39 week gestational age, n = 12) were exposed to cyclosporine A (CSA, 0, 62.5, 125, 1250 ng/mL) or azathioprine (AZA, 0, 5, 10, 100 ng/mL) separately or, in combination for up to 48 hours. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays showed a significant decrease in the explant metabolic activity between AZA and the control group (24 hours, 100 ng/mL, 48 hours, all concentrations, P < .005). Cyclosporin A (CsA) reduced cell activity when associated with AZA (48 hours, P < .005). Fibrinoid deposits increased in AZA-treated explants alone (5 ng/mL, 48 hours; 10 ng/mL, 24-48 hours; P < .005) or when associated with CsA (10 AZA/125 CsA, P < .05), whereas in CsA treatment alone, there was an augment in syncytial knots (24-48 hours, P < .005). The sFLT1 gene (24 hours, P < .05) and protein (P < .005) expression increased in AZA and CsA-treatments separately or in combination (P < .05). Placental growth factor increased in AZA (24 hours, 10 ng/mL) and CsA (125 ng/mL; P < .05). In conclusion, our data indicate that AZA primarily acts on the villous metabolism, perturbing placental homeostasis. Since these drugs may alter the balance of angiogenic factors in its selection for clinical application, their impact on the behavior of placental villous should be considered.
Asunto(s)
Azatioprina/farmacología , Ciclosporina/farmacología , Inmunosupresores/farmacología , Placenta/efectos de los fármacos , Vellosidades Coriónicas/efectos de los fármacos , Vellosidades Coriónicas/metabolismo , Femenino , Humanos , Placenta/metabolismo , Factor de Crecimiento Placentario/metabolismo , Embarazo , Receptor 1 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Circular RNAs (circRNAs) have recently been shown to exert their effects on multiple pathological processes by acting as microRNA (miRNA) sponges. However, the roles of circRNAs in gestational diabetes mellitus (GDM) are largely unknown. This study aimed to identify the circRNAs involved in GDM and predict their potential biological functions. We first performed next-generation sequencing (NGS) to generate unbiased placental villi circRNA expression profiles of GDM and normal controls. In total, 48,270 circRNAs from the placental villi of the two groups were sequenced. Of these, 227 circRNAs were significantly up-regulated and 255 circRNAs were significantly down-regulated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) biological pathway analyses demonstrated that glycometabolism and lipometabolism processes, which are important in GDM development, were significantly enriched. Further analysis showed that most of the circRNAs harbored miRNA binding sites, and some were associated with GDM. These results showed that circRNAs are aberrantly expressed in the placental villi of GDM patients and play potential roles in the development of GDM.
Asunto(s)
Vellosidades Coriónicas/metabolismo , Diabetes Gestacional/genética , Perfilación de la Expresión Génica , ARN/genética , Adulto , Vellosidades Coriónicas/patología , Diabetes Gestacional/patología , Regulación hacia Abajo , Femenino , Ontología de Genes , Redes Reguladoras de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , MicroARNs/genética , Embarazo , ARN/análisis , ARN Circular , Regulación hacia ArribaRESUMEN
Placental barrier regulates maternal-fetal interchange protecting the baby from damage caused by substances found in the uterine environment or circulating in the vascular system. Organophosphate (OP) pesticides are a paramount group of environmental pollutants used in intensive agriculture for protection against diseases and pests. While many studies have reported an increased risk of pregnancy alterations in pregnant women exposed to OPs, few have analyzed the effects caused by these pesticides in the placenta. Herein, we evaluated the effects of chlorpyrifos (CPF), one of the most widely used OP insecticides, on human placenta using in vitro and ex vivo exposure models. Villous cytotrophoblast cells isolated from normal human term placentas maintained their cell viability, differentiated into syncytiotrophoblast-like structures, and increased the expression of ß-hCG, ABCG2, and P-gp in the presence of CPF at concentrations of 10 to 100µM. The same doses of CPF induced marked changes in chorionic villi samples. Indeed, CPF exposure increased stroma cell apoptosis, altered villi matrix composition, basement membrane thickness, and trophoblastic layer integrity. Histomorphological and ultrastructural alterations are compatible with those found in placentas where maternal-placenta injury is chronic and able to impair the placental barrier function and nutrient transport from mother to the fetus. Our study shows that placental ex vivo exposure to CPF produces tissue alterations and suggest that human placenta is a potential target of CPF toxicity. In addition, it highlights the importance of using different models to assess the effects of a toxic on human placenta.
Asunto(s)
Cloropirifos/toxicidad , Inhibidores de la Colinesterasa/toxicidad , Vellosidades Coriónicas/efectos de los fármacos , Insecticidas/toxicidad , Trofoblastos/efectos de los fármacos , Subfamilia B de Transportador de Casetes de Unión a ATP/metabolismo , Transportador de Casetes de Unión a ATP, Subfamilia G, Miembro 2/metabolismo , Apoptosis/efectos de los fármacos , Membrana Basal/efectos de los fármacos , Membrana Basal/ultraestructura , Bioensayo , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Gonadotropina Coriónica Humana de Subunidad beta/metabolismo , Vellosidades Coriónicas/metabolismo , Vellosidades Coriónicas/ultraestructura , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Proteínas de Neoplasias/metabolismo , Embarazo , Reproducibilidad de los Resultados , Medición de Riesgo , Células del Estroma/efectos de los fármacos , Células del Estroma/ultraestructura , Factores de Tiempo , Técnicas de Cultivo de Tejidos , Pruebas de Toxicidad/métodos , Trofoblastos/metabolismo , Trofoblastos/ultraestructuraRESUMEN
This chapter discusses the relationship between failure in placentation and the subsequent alterations in the normal structure of the placenta. Interstitial Cajal-like cells (ICLC) were observed for the first time in the human placenta in 2007 and later were named telocytes. Strong evidence confirms that in the placental chorionic villi, TC are located strategically between the smooth muscle cells (SMC) of the fetal blood vessel wall and the stromal myofibroblasts. As the placenta is a non-innervated organ and considering the strategic position of telocytes in chorionic villi, it has been postulated that their function would be related to signal transduction mechanisms involved in the regulation of the blood flow in the fetal vessels, as well as in the shortening/lengthening of the chorionic villi providing the necessary rhythmicity to the process of maternal/fetal metabolic exchange. In this context, telocytes represent part of a functional triad: "SMC of fetal blood vessel-telocyte-myofibroblast." This triad takes part in the regulation of fetal growth and development via transport of nutrients and gases. This chapter also discusses the alterations in the metabolic maternal-fetal exchange, leading to intrauterine growth retardation and preeclampsia. Additionally, the apoptosis undergoing in the preeclamptic hypoxic placenta affects all the chorionic villi cells, including telocytes and myofibroblast, and not only trophoblast, as it has been so far considered. In consequence, we proposed that apoptosis affects the triad structure and alters the placental function, subsequently affecting the normal fetal growth and development.
Asunto(s)
Retardo del Crecimiento Fetal/patología , Hipoxia/patología , Miocitos del Músculo Liso/patología , Miofibroblastos/patología , Preeclampsia/patología , Telocitos/patología , Apoptosis , Vellosidades Coriónicas/metabolismo , Vellosidades Coriónicas/patología , Femenino , Retardo del Crecimiento Fetal/metabolismo , Humanos , Hipoxia/metabolismo , Intercambio Materno-Fetal , Miocitos del Músculo Liso/metabolismo , Miofibroblastos/metabolismo , Preeclampsia/metabolismo , Embarazo , Transducción de Señal , Telocitos/metabolismo , Trofoblastos/metabolismo , Trofoblastos/patologíaRESUMEN
Habitual abortion is associated with the altered expression of multiple genes. This study was carried out to investigate the relationship between expression of Toll-like receptor 4 (TLR4) and monocyte chemotactic protein 2 (MCP2 or CCL8) and habitual abortion. This was done by detecting and comparing their relative expression in peripheral blood and placental villi of patients and healthy fertile women. Based on our previous research, 85 subjects with habitual abortion (study group) and 40 healthy fertile women (control group), who were admitted to our hospital between June 2013 and December 2014, were enrolled in this study. After these subjects signed written informed consent, peripheral blood samples and villous tissues were collected, from which the total RNA was extracted. The expression of TLR4 and MCP2 was detected with quantitative reverse transcription-polymerase chain reaction, using GAPDH as a reference control. The expression of TLR4 and MCP2 in the peripheral blood and villous tissues of the study group was significantly higher than that of the control group (P < 0.05). A positive correlation was also observed between the changes in expression levels of TLR4 and MCP2. In conclusion, TLR4 and MCP2 expression correlated with the occurrence of habitual abortion. Detecting expression changes in TLR4 and MCP2 in the peripheral blood is a feasible method for predicting the occurrence of abortion in women of child-bearing age.
Asunto(s)
Aborto Habitual/genética , Quimiocina CCL8/biosíntesis , Receptor Toll-Like 4/biosíntesis , Aborto Habitual/patología , Adulto , Quimiocina CCL8/genética , Vellosidades Coriónicas/metabolismo , Femenino , Regulación de la Expresión Génica , Humanos , Embarazo , Receptor Toll-Like 4/genéticaRESUMEN
Omentin-1 is an adipocytokine with anti-inflammatory activity that has been associated with different metabolic disorders. The aim of this study is to investigate the serum profiles of omentin-1 throughout human and rat pregnancy. Serum omentin-1 levels were determined by ELISA in a prospective cohort study of healthy pregnant women (n=40) during the three trimesters of pregnancy and in twenty healthy non-pregnant women during the follicular and luteal phase of the menstrual cycle. In addition, serum omentin-1 levels were measured in rats during different periods of pregnancy (gestational days 8, 12, 16, 19, and 21) and in an age-matched control (virgin) group of rats (n=12rats/group). Finally, immunohistochemistry was used to demonstrate the presence of omentin-1 protein in human and rat placenta. Omentin-1 immunoreactivity was detected in cytotrophoblasts, syncytiotrophoblasts, sparse Hofbauer cells, and endothelial cells of the stem villi of human placenta. Additionally, it was detected in the labyrinthine trophoblast and yolk sac layer of the rat placenta. Human and rat serum omentin-1 levels were significantly lower in the late gestational period when compared with the non-pregnant women and virgin rats (p<0.05). Serum omentin-1 changes were not significant throughout the gestation in both species (p>0.05). Human serum omentin-1 levels have an inverse relationship with triglyceride levels during pregnancy. Our findings have not determined the exact role of omentin-1 during pregnancy, concerning the metabolic control of triglycerides and other energy sources. Whether omentin-1 decrease implies a regulatory function is still not clear. Further studies are needed to address this issue and determine the role of omentin-1 in metabolic adaptations during normal human and rat pregnancy.
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Citocinas/sangre , Regulación del Desarrollo de la Expresión Génica , Lectinas/sangre , Adulto , Animales , Vellosidades Coriónicas/metabolismo , Modelos Animales de Enfermedad , Células Endoteliales/metabolismo , Ensayo de Inmunoadsorción Enzimática , Femenino , Proteínas Ligadas a GPI/sangre , Humanos , Inmunohistoquímica , Embarazo , Preñez , Ratas , Ratas Sprague-Dawley , Factores de Tiempo , Triglicéridos/metabolismo , Trofoblastos/metabolismo , Saco Vitelino/metabolismo , Adulto JovenRESUMEN
Cell-cell fusion is an essential event during life. Throughout human pregnancy, the syncytiotrophoblast (STB) layer of the placenta is formed by continuous fusion of the underlying villous cytotrophoblasts, thus maintaining placental functionality. Defects in this process are associated with pathologies like pre-eclampsia and intrauterine growth restriction. Krüppel-like factor 6 (KLF6) is a transcription factor highly expressed in human and murine placenta. However, KLF6 functions in trophoblast cells remain largely unexplored. The aim of this work was to address the role of KLF6 during STB formation. KLF6 knockdown through small interfering RNA experiments hindered cell-cell fusion revealed by immunofluorescence microscopy in human primary villous cytotrophoblast as well as in the human placental-derived BeWo cell line. Furthermore, KLF6 silencing led to a decrease in the expression of the fusogenic protein Syncytin-1 and the cell cycle regulator p21 CIP1/WAF1: measured by quantitative RT-PCR and western blot assays. On the contrary, transcript levels of genes that encode for proteins involved in STB formation such as Syncytin-1, Syncytin-2, Connexin-43 and Zonula Occludens-1 increased when KLF6 was overexpressed in differentiating villous cytotrophoblasts and in non-fusing placental-derived JEG-3 cells. Interestingly, the expression of two trophoblast biochemical differentiation markers, ßhCG and PSG3, were not reduced after KLF6 silencing in differentiating trophoblast cells. Present results support the notion that KLF6 is a relevant participant in cytotrophoblast fusion.
Asunto(s)
Vellosidades Coriónicas/metabolismo , Factores de Transcripción de Tipo Kruppel/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Trofoblastos/metabolismo , Adulto , Fusión Celular , Gonadotropina Coriónica Humana de Subunidad beta/genética , Gonadotropina Coriónica Humana de Subunidad beta/metabolismo , Vellosidades Coriónicas/crecimiento & desarrollo , Conexina 43/genética , Conexina 43/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Femenino , Regulación de la Expresión Génica , Productos del Gen env/genética , Productos del Gen env/metabolismo , Humanos , Factor 6 Similar a Kruppel , Factores de Transcripción de Tipo Kruppel/antagonistas & inhibidores , Factores de Transcripción de Tipo Kruppel/genética , Embarazo , Proteínas Gestacionales/genética , Proteínas Gestacionales/metabolismo , Cultivo Primario de Células , Proteínas Proto-Oncogénicas/antagonistas & inhibidores , Proteínas Proto-Oncogénicas/genética , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Trofoblastos/citología , Proteína de la Zonula Occludens-1/genética , Proteína de la Zonula Occludens-1/metabolismoRESUMEN
With the advancement of ultrasonic technology in recent years, sonography has become a common medical diagnostic tool, as it has elevated output sonic intensity and elongated exposure time. This study investigates the effect of ultrasound on human chorionic villus cell apoptosis during early pregnancy. Transvaginal ultrasound was performed for a total of 60 women who had undergone induced abortion at our hospital. They were randomly divided into the control, short ultrasound (10 min), and long ultrasound (20 min) groups (N = 20 each). Twenty-four hours after ultrasonic exposure, chorionic villus tissues were extracted during induced abortion, and were tested for cell apoptosis using flow cytometry. Bax and B cell lymphoma-2 (Bcl-2) protein levels were also quantified by immunohistochemistry. We found that the long ultrasound group had significantly higher cell apoptosis rates compared to the short ultrasound group, which in turn had higher rates compared to the control group (P < 0.05 in both cases). Bax protein levels were elevated in both the long and short ultrasound groups (P < 0.05). Bcl-2 proteins in two ultrasound groups, however, were downregulated as compared to those in the control group (P < 0.05). It is therefore possible that transvaginal sonography can potentiate the apoptosis of human chorionic villus cells by increasing the Bax/Bcl-2 protein ratio.
Asunto(s)
Apoptosis/efectos de la radiación , Vellosidades Coriónicas/diagnóstico por imagen , Proteínas Proto-Oncogénicas c-bcl-2/genética , Ondas Ultrasónicas/efectos adversos , Proteína X Asociada a bcl-2/genética , Aborto Inducido , Adulto , Vellosidades Coriónicas/metabolismo , Femenino , Expresión Génica , Humanos , Inmunohistoquímica , Embarazo , Primer Trimestre del Embarazo , Proteínas Proto-Oncogénicas c-bcl-2/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Factores de Tiempo , Ultrasonografía , Vagina/diagnóstico por imagen , Proteína X Asociada a bcl-2/agonistas , Proteína X Asociada a bcl-2/metabolismoRESUMEN
INTRODUCTION: ICAM-1 expression on the villous syncytiotrophoblast (ST) is believed to participate in migration of maternal cells into the inflamed villi regardless of villitis etiology. However, its expression on immune cells in chronic villitis (CV) has yet to be analyzed. ICAM-1 induces cell-cell adhesion allowing intercellular communication, T cell-mediated defense mechanism, and inflammatory response. MATERIAL AND METHODS: 21 cases of CV (all without an identifiable etiologic agent) and 3 control placentas were analyzed using ICAM-1, and for immune cells CD45, CD3 and CD68. These cells were subdivided according to their location in inflamed villi: a) within the inflamed villi and b) outside forming perivillous aggregates. RESULTS: Large amounts of CD45, CD3 and CD68 were found within the inflamed villi and forming perivillous aggregates attached to areas of trophoblastic loss. Inflamed villi usually showed ICAM-1+ ST. The majority of immune cells surrounding areas of trophoblastic rupture presented marked expression of ICAM-1. In contrast, a small number of immune cells within the inflamed villi exhibited ICAM-1 expression. Only some (<5%) inflamed villi without trophoblastic rupture and with ICAM-1+ ST presented adherence of immune cells. DISCUSSION: In inflamed villi of chronic villitis, the level of ICAM-1 expression on immune cells depends on their location: high in number of cells in the perivillous region and low within the villi. The strongest expression of ICAM-1 on immune cells attached to areas of trophoblastic rupture suggests that the loss of trophoblast can lead to an amplification of the inflammatory response.
Asunto(s)
Vellosidades Coriónicas/metabolismo , Inflamación/metabolismo , Molécula 1 de Adhesión Intercelular/metabolismo , Enfermedades Placentarias/metabolismo , Vellosidades Coriónicas/inmunología , Femenino , Humanos , Inflamación/inmunología , Linfocitos/inmunología , Linfocitos/metabolismo , Macrófagos/inmunología , Macrófagos/metabolismo , Monocitos/inmunología , Monocitos/metabolismo , Enfermedades Placentarias/inmunología , EmbarazoRESUMEN
Programs for the prevention of mother-to-child transmission of HIV have reduced the transmission rate of perinatal HIV infection and have thereby increased the number of HIV-exposed uninfected (HEU) infants. Natural immunity to HIV-1 infection in both mothers and newborns needs to be further explored. In this study, we compared the expression of antiviral restricting factors in HIV-infected pregnant mothers treated with antiretroviral therapy (ART) in pregnancy (n=23) and in cord blood (CB) (n=16), placental tissues (n=10-13) and colostrum (n=5-6) samples and compared them to expression in samples from uninfected (UN) pregnant mothers (n=21). Mononuclear cells (MNCs) were prepared from maternal and CB samples following deliveries by cesarean section. Maternal (decidua) and fetal (chorionic villus) placental tissues were obtained, and colostrum was collected 24 h after delivery. The mRNA and protein expression levels of antiviral factors were then evaluated. We observed a significant increase in the mRNA expression levels of antiviral factors in MNCs from HIV-infected mothers and CB, including the apolipoprotein B mRNA-editing enzyme 3G (A3G), A3F, tripartite motif family-5α (TRIM-5α), TRIM-22, myxovirus resistance protein A (MxA), stimulator of interferon (IFN) genes (STING) and IFN-ß, compared with the levels detected in uninfected (UN) mother-CB pairs. Moreover, A3G transcript and protein levels and α-defensin transcript levels were decreased in the decidua of HIV-infected mothers. Decreased TRIM-5α protein levels in the villi and increased STING mRNA expression in both placental tissues were also observed in HIV-infected mothers compared with uninfected (UN) mothers. Additionally, colostrum cells from infected mothers showed increased tetherin and IFN-ß mRNA levels and CXCL9 protein levels. The data presented here indicate that antiviral restricting factor expression can be induced in utero in HIV-infected mothers. Future studies are warranted to determine whether this upregulation of antiviral factors during the perinatal period has a protective effect against HIV-1 infection.
Asunto(s)
Sangre Fetal/metabolismo , Regulación de la Expresión Génica/inmunología , Infecciones por VIH/sangre , Infecciones por VIH/inmunología , Inmunidad Innata/inmunología , Viremia/prevención & control , Desaminasa APOBEC-3G , Factores de Restricción Antivirales , Western Blotting , Brasil , Proteínas Portadoras/metabolismo , Vellosidades Coriónicas/metabolismo , Calostro/metabolismo , Citidina Desaminasa/metabolismo , Cartilla de ADN/genética , Decidua/metabolismo , Femenino , Perfilación de la Expresión Génica , Humanos , Interferón beta/economía , Interferón beta/metabolismo , Leucocitos Mononucleares/metabolismo , Proteínas de la Membrana/metabolismo , Antígenos de Histocompatibilidad Menor , Madres , Proteínas de Resistencia a Mixovirus/metabolismo , Embarazo , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteínas Represoras/metabolismo , Estadísticas no Paramétricas , Proteínas de Motivos Tripartitos , Ubiquitina-Proteína Ligasas , Viremia/metabolismoRESUMEN
The aim of this study is to determine the concentrations of lidocaine and its metabolite, monoethylglycine xylidide (MEGX), and of the enantiomers of bupivacaine in maternal and fetal compartments. Ten healthy pregnant women were submitted to epidural anesthesia. Drug concentrations were determined in the maternal vein, fetal umbilical artery and vein, and the placental intervillous space. The highest concentrations of the bupivacaine enantiomers lidocaine and of lidocaine and of its MEGX metabolite were detected in maternal plasma and in the placental intervillous space. The placental transfer was 33% for the (+)-(R)-bupivacaine enantiomer and 31% for the (-)-(S)-bupivacaine enantiomer. For lidocaine and its MEGX metabolite, respective placental transfers were 60% and 43%. Lidocaine concentration in the fetal umbilical vein was 1.46 times higher than in the fetal umbilical artery. The highest concentrations of lidocaine and its metabolite and of the enantiomers of bupivacaine were detected in the placental intervillous space. The higher lidocaine concentrations in the fetal umbilical vein than in the fetal umbilical artery suggest that there was tissue uptake of the drug or drug metabolization by the fetus.
Asunto(s)
Anestésicos Locales/farmacocinética , Bupivacaína/farmacocinética , Lidocaína/análogos & derivados , Lidocaína/farmacocinética , Adulto , Anestesia Epidural/métodos , Anestesia Obstétrica/métodos , Anestésicos Locales/química , Bupivacaína/química , Vellosidades Coriónicas/metabolismo , Femenino , Humanos , Placenta/metabolismo , Embarazo , Estereoisomerismo , Distribución Tisular , Arterias Umbilicales/metabolismo , Venas Umbilicales/metabolismo , Adulto JovenRESUMEN
BACKGROUND: Fentanyl is used in obstetrical practice to promote analgesia and anesthesia during labor and in cesarean delivery, with rapid and short-term effects. OBJECTIVE: To determine fentanyl concentrations in maternal plasma, in the placental intervillous space, and in the umbilical artery and vein in term pregnant women. PATIENTS AND METHODS: Ten healthy pregnant women underwent epidural anesthesia with fentanyl plus bupivacaine and lidocaine, and fentanyl concentrations were determined in the various maternal and fetal compartments, including the placental intervillous space, which has not been previously studied in the literature. RESULTS: The ratios of fentanyl concentrations in the various maternal and fetal compartments revealed an 86% rate of placental fentanyl transfer. The highest fentanyl concentrations were detected in the placental intervillous space, being 2.19 times higher than in maternal plasma, 2.8 times higher than in the umbilical vein and 3.6 times higher than in the umbilical artery, with no significant differences between the umbilical vein and artery, demonstrating that there was no drug uptake by fetal tissues nor metabolism of the drug by the fetus despite the high rates of placental transfer. CONCLUSION: The present study demonstrated that the placental intervillous space acted as a site of fentanyl deposit, a fact that may be explained by two hypotheses: (1) the blood collected from the placental intervillous space is arterial and, according to some investigators, the arterial plasma concentrations of the drugs administered to patients undergoing epidural anesthesia are higher than the venous concentrations, and (2) a possible role of P-glycoprotein (P-gp).
Asunto(s)
Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Analgésicos Opioides/farmacocinética , Anestésicos Intravenosos/farmacocinética , Fentanilo/farmacocinética , Feto/metabolismo , Placenta/metabolismo , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/efectos de los fármacos , Adulto , Analgesia Obstétrica/métodos , Analgésicos Opioides/administración & dosificación , Analgésicos Opioides/sangre , Anestesia Obstétrica/métodos , Anestésicos Intravenosos/administración & dosificación , Anestésicos Intravenosos/sangre , Vellosidades Coriónicas/metabolismo , Femenino , Fentanilo/administración & dosificación , Fentanilo/sangre , Humanos , EmbarazoRESUMEN
It is known that human syncytiotrophoblast (hSCT) actively transports more than 80% of the Ca2+ that goes from maternal to fetal circulation. Transepithelial transport of Ca2+ is carried out through channels, transporters and exchangers located in both microvillous (MVM) and basal (BM) plasma membranes. The plasma membrane Ca-ATPase (PMCA) is the most important mechanism of Ca2+ homeostasis control in the human placenta. In this work, we reexamined the distribution of PMCA in isolated hSCT of term placenta. The PMCA activity was determined in isolated hSCT plasma membranes. A partial characterization of the PMCA activity was performed, including an evaluation of the sensitivity of this enzyme to an in vitro induced lipid peroxidation. Expression of the PMCA in hSCT plasma membranes and tissue sections was investigated using Western blots and immunohistochemistry, respectively. Our study demonstrates, for the first time, a correlation between the activity and structural distribution of PMCA in both MVM and BM of hSCT. It also demonstrates a higher PMCA activity and expression in MVM as compared to BM. Finally, PMCA4 seems to be preferentially distributed in both hSCT plasma membranes, while PMCA1 is shown to be present in the hSCT homogenate. However, the membrane fractions did not show any PMCA1 labeling. Our results must be taken into account in order to propose a new model for the transport of calcium across the hSCT.
Asunto(s)
Membrana Celular/metabolismo , Vellosidades Coriónicas/metabolismo , ATPasas Transportadoras de Calcio de la Membrana Plasmática/metabolismo , Nacimiento a Término/metabolismo , Trofoblastos/metabolismo , Transporte Biológico/fisiología , Calcio/metabolismo , Separación Celular , Vellosidades Coriónicas/enzimología , Vellosidades Coriónicas/ultraestructura , Femenino , Humanos , Isoenzimas/metabolismo , Microvellosidades/metabolismo , ATPasas Transportadoras de Calcio de la Membrana Plasmática/química , ATPasas Transportadoras de Calcio de la Membrana Plasmática/fisiología , Embarazo , Distribución Tisular , Extractos de Tejidos/química , Extractos de Tejidos/metabolismo , Trofoblastos/enzimología , Trofoblastos/ultraestructuraRESUMEN
BACKGROUND: Several factors determine the risk of HIV mother-to-child transmission (MTCT), such as coinfections in placentas from HIV-1 positive mothers with other pathogens. Chagas' disease is one of the most endemic zoonoses in Latin America, caused by the protozoan Trypanosoma cruzi. The purpose of the study was to determine whether T. cruzi modifies HIV infection of the placenta at the tissue or cellular level. RESULTS: Simple and double infections were carried out on a placental histoculture system (chorionic villi isolated from term placentas from HIV and Chagas negative mothers) and on the choriocarcinoma BeWo cell line. Trypomastigotes of T. cruzi (VD lethal strain), either purified from mouse blood or from Vero cell cultures, 24 h-supernatants of blood and cellular trypomastigotes, and the VSV-G pseudotyped HIV-1 reporter virus were used for the coinfections. Viral transduction was evaluated by quantification of luciferase activity. Coinfection with whole trypomastigotes, either from mouse blood or from cell cultures, decreased viral pseudotype luciferase activity in placental histocultures. Similar results were obtained from BeWo cells. Supernatants of stimulated histocultures were used for the simultaneous determination of 29 cytokines and chemokines with the Luminex technology. In histocultures infected with trypomastigotes, as well as in coinfected tissues, IL-6, IL-8, IP-10 and MCP-1 production was significantly lower than in controls or HIV-1 transducted tissue. A similar decrease was observed in histocultures treated with 24 h-supernatants of blood trypomastigotes, but not in coinfected tissues. CONCLUSION: Our results demonstrated that the presence of an intracellular pathogen, such as T. cruzi, is able to impair HIV-1 transduction in an in vitro system of human placental histoculture. Direct effects of the parasite on cellular structures as well as on cellular/viral proteins essential for HIV-1 replication might influence viral transduction in this model. Nonetheless, additional mechanisms including modulation of cytokines/chemokines at placental level could not be excluded in the inhibition observed. Further experiments need to be conducted in order to elucidate the mechanism(s) involved in this phenomenon. Therefore, coinfection with T. cruzi may have a deleterious effect on HIV-1 transduction and thus could play an important role in viral outcome at the placental level.
Asunto(s)
Enfermedad de Chagas/virología , Vellosidades Coriónicas/virología , VIH-1/fisiología , Trypanosoma cruzi/fisiología , Animales , Línea Celular , Enfermedad de Chagas/patología , Enfermedad de Chagas/fisiopatología , Vellosidades Coriónicas/anatomía & histología , Vellosidades Coriónicas/metabolismo , Femenino , VIH-1/metabolismo , VIH-1/patogenicidad , Humanos , Placenta/inmunología , Placenta/virología , Replicación Viral/efectos de los fármacosRESUMEN
In an effort to define the varied expression of three vasoactive markers in the clinical models of normal placenta/ normal invasion (n = 11), preeclampsia/restricted trophoblast invasion (n = 15), and placenta accreta/exaggerated invasion (n = 6), we performed semiquantitative immunohistochemistry for kallikrein, bradykinin B2 receptor, and endothelial nitric oxide synthase (eNOS). In the floating villi, the syncytiotrophoblast expressed more kallikrein in placenta accreta (p < 0.05), than in normal and preeclamptic placentas, while the bradykinin B2 receptor and eNOS were similarly expressed in all groups; in the fetal endothelium, the bradykinin B2 receptor was enhanced in placenta accreta (p < 0.005), but kallikrein and eNOS were similarly expressed in the other two groups. In the extravillous trophoblast, both kallikrein and eNOS expression were higher in placenta accreta (p < 0.001), while the bradykinin B2 receptor signal was only enhanced in preeclampsia (p < 0.05). The presence and localization of kallikrein, the bradykinin B2 receptor, and eNOS in the fetomaternal interface in the three study conditions supports a local role for interrelated vasodilatory/antiaggregating systems. This first report of the variations observed in kallikrein and eNOS in a condition of exaggerated trophoblast invasion supports the participation of vasodilatation in trophoblast migration.
Asunto(s)
Calicreínas/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Placenta Accreta/metabolismo , Placenta/metabolismo , Preeclampsia/metabolismo , Receptor de Bradiquinina B2/metabolismo , Vellosidades Coriónicas/metabolismo , Células Endoteliales/metabolismo , Femenino , Feto/metabolismo , Expresión Génica , Edad Gestacional , Humanos , Calicreínas/orina , Intercambio Materno-Fetal , Modelos Biológicos , Placenta/irrigación sanguínea , Embarazo , Complicaciones Hematológicas del Embarazo/metabolismo , Resultado del Embarazo , Trofoblastos/metabolismoRESUMEN
Although an in vitro study has hypothesized that expression of ICAM-1 by villous trophoblasts could be important for the influx of maternal immune cells in villitis, it remains to be shown whether the same phenomenon occurs in human villitis. To investigate the expression of ICAM-1 by villous trophoblasts, its relationship with rupture of the trophoblastic barrier and influx of immune cells into the villi, we analysed 18 paraffin-embedded placentas with placentitis (5 by Toxoplasma gondii, 3 by Trypanosoma cruzi, 2 by Paracoccidioides brasiliensis and 8 of unknown aetiology - VUE) and 8 control placentas for detection of ICAM-1 by immunohistochemistry. All cases but one of placentitis showed trophoblast overexpression of ICAM-1 in the inflamed villi, located almost exclusively next to the areas of trophoblastic rupture. The villitis cases (caused by T. cruzi, T. gondii and VUE) presented leukocyte adherence in the areas of trophoblastic rupture. When the inflammatory reaction was situated in the intervillous space (placentitis by P. brasiliensis), in spite of the trophoblastic rupture and ICAM-1 overexpression there was no leukocyte influx into villi. None of the control placentas showed ICAM-1 expression by the trophoblast. We concluded that overexpression of ICAM-1 by villous trophoblasts occurs during placentitis characterized by accumulation of leukocytes in the villous or intervillous space and probably plays an important role in the rupture of the trophoblastic barrier. The influx of immune cells into the villi appears to be mediated by ICAM-1 but the location of the antigen within villous stroma is certainly a crucial factor for its occurrence.
Asunto(s)
Vellosidades Coriónicas/metabolismo , Molécula 1 de Adhesión Intercelular/metabolismo , Enfermedades Placentarias/metabolismo , Trofoblastos/metabolismo , Adulto , Biomarcadores/metabolismo , Enfermedad de Chagas/complicaciones , Enfermedad de Chagas/metabolismo , Enfermedad de Chagas/patología , Vellosidades Coriónicas/patología , Femenino , Humanos , Inmunohistoquímica , Inflamación/metabolismo , Inflamación/patología , Leucocitos/metabolismo , Leucocitos/patología , Enfermedades Placentarias/parasitología , Enfermedades Placentarias/patología , Embarazo , Toxoplasmosis/complicaciones , Toxoplasmosis/metabolismo , Toxoplasmosis/patología , Trofoblastos/patologíaRESUMEN
Maternal infection of Trypanosoma cruzi is associated with premature births, abortions and placentitis. A decrease in EGF levels has been suggested to occur in animals infected by T. cruzi, but there is no research about the levels of EGF in human patients with Chagas' disease. We evaluated serum EGF levels in pregnant women with and without the disease, and with immunological methods detected EGF receptors and EGF in both groups of placentae and in cultures of normal placental villi with and without parasites. PLAP in placentae from those women was also immunologically detected, since EGF can induce the release of PLAP from the trophoblast surface and PLAP is suggested to be a receptor allowing parasite invasion of the placenta. Plasma from women with Chagas' disease contained lower level of EGF when compared to plasma of healthy women. Placentae from women with Chagas' disease showed lower PLAP expression but same level of detectable EGF receptors and EGF when compared with placentae from women without the disease. Culture with parasites did not reduce EGFr level. Results suggest a lower availability of EGF in women with Chagas' disease, which could explain several malfunctions of the placenta associated with maternal Chagas' disease.