RESUMEN
The migration, proliferation, and apoptosis of trophoblastic cells play a crucial role in ensuring the effective preservation of pregnancy at the maternal-fetal interface. Any deviations in the structure and function of these cells might potentially result in the development of numerous pregnancy-related disorders, including missed abortion (MA). This study involved the examination of semaphorin 4A (SEMA4A) expression in missed abortion (n = 18) and normal early pregnancy (n = 18) villus. The findings of this study indicate a statistically significant decrease in the expression of SEMA4A in the villi of individuals diagnosed with missed abortion, as compared to the control group. The results of our vitro study showed that SEMA4A promoted the migration and proliferation of trophoblast cells and inhibited their apoptosis. Subsequent studies have shown that SEMA4A may be involved in regulating p-STAT3/STAT3, MMP9, bcl-2, and BAX levels. In summary, the findings of this study indicate a correlation between the decreased level of SEMA4A in chorionic villi and missed abortion. These results offer novel theoretical insights into the proper implantation and development of SEMA4A embryos at the maternal-fetal interface.
Asunto(s)
Apoptosis , Proliferación Celular , Factor de Transcripción STAT3 , Semaforinas , Transducción de Señal , Trofoblastos , Humanos , Femenino , Trofoblastos/metabolismo , Embarazo , Semaforinas/metabolismo , Semaforinas/genética , Factor de Transcripción STAT3/metabolismo , Adulto , Movimiento Celular , Vellosidades Coriónicas/metabolismo , Aborto Retenido/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismoRESUMEN
The aim of this study was to evaluate the role of angiotensin-converting enzyme 1 (ACE1) in H2O2-induced trophoblast cell injury and the potential molecular mechanisms. Oxidative stress was modeled by exposing HTR-8/SVneo cells to 200 µM H2O2. Western blot and real-time quantitative PCR methods were used to detect protein and mRNA expression level of ACE1 in chorionic villus tissue and trophoblast HTR-8/SVneo cell. Inhibition of ACE1 expression was achieved by transfection with small interfering RNA. Then flow cytometry, Cell Counting Kit-8, and Transwell assay was used to assess apoptosis, viability, and migration ability of the cells. Reactive oxygen species (ROS) were detected by fluorescent probes, and malondialdehyde (MDA), superoxide dismutase (SOD), and reduced glutathione (GSH) activities were determined by corresponding detection kits. Angiotensin-converting enzyme 1 expression was upregulated in chorionic villus tissue of patients with missed abortion (MA) compared with individuals with normal early pregnancy abortion. H2O2 induced elevated ACE1 expression in HTR-8/SVneo cells, promoted apoptosis, and inhibited cell viability and migration. Knockdown of ACE1 expression inhibited H2O2-induced effects to enhance cell viability and migration and suppress apoptosis. Additionally, H2O2 stimulation caused increased levels of ROS and MDA and decreased SOD and GSH activity in the cells, whereas knockdown of ACE1 expression led to opposite changes of these oxidative stress indicators. Moreover, knockdown of ACE1 attenuated the inhibitory effect of H2O2 on the Nrf2/HO-1 pathway. Angiotensin-converting enzyme 1 was associated with MA, and it promoted H2O2-induced injury of trophoblast cells through inhibiting the Nrf2 pathway. Therefore, ACE1 may serve as a potential therapeutic target for MA.
Asunto(s)
Aborto Retenido , Peróxido de Hidrógeno , Peptidil-Dipeptidasa A , Trofoblastos , Humanos , Trofoblastos/metabolismo , Trofoblastos/efectos de los fármacos , Peróxido de Hidrógeno/farmacología , Peptidil-Dipeptidasa A/genética , Peptidil-Dipeptidasa A/metabolismo , Femenino , Embarazo , Aborto Retenido/genética , Aborto Retenido/metabolismo , Apoptosis/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Línea Celular , Especies Reactivas de Oxígeno/metabolismo , Supervivencia Celular/efectos de los fármacos , Adulto , Movimiento Celular/efectos de los fármacosRESUMEN
OBJECTIVE: To investigate the expression of Siglec10 and CD24 in normal early pregnancy and missed abortion, and their significance in the maternal-fetal interface. METHODS: For our research, we employed Q-PCR and WB techniques to evaluate the traits and expression of Siglec10 and CD24 in the nonpregnant endometrium, as well as in the villus and decidua of women in their 6-10 weeks of normal early pregnancy and those who experienced missed abortion. Additionally, we utilized ELISA to determine the levels of Siglec10 and CD24 in the peripheral blood of pregnancy, missed abortion, and non-pregnant individuals. T-test and ANOVA were used to compare groups. RESULTS: 1. Villous tissues in early pregnancy showed high expression of Siglec10 and CD24, with a significant increase in expression in the missed abortion group (P < 0.01).2. Nonpregnant endometrial tissue showed low expression of Siglec10 and CD24, while early pregnancy decidua showed high expression, with even higher expression in missed abortion (all P < 0.05).3. Serum levels of Siglec10 and CD24 in normal early pregnancy were significantly higher than non-pregnancy (P < 0.01). However, the missed abortion group showed significantly higher levels than normal pregnancy (P < 0.01).4. CD24 expression in serum of missed abortion increases with Siglec10 expression, indicating a significant positive correlation (r = 0.500, P < 0.01). CONCLUSION: Siglec10 and CD24 expression in villus, decidua, and peripheral blood are up-regulated in unexplained missed abortions than those of women with normal pregnancies. This suggests that the levels of serum Siglec10 and CD24 can be used as an effective predictor of missed abortion.
Asunto(s)
Aborto Retenido , Femenino , Humanos , Embarazo , Aborto Retenido/genética , Aborto Retenido/metabolismo , Antígeno CD24/genética , Antígeno CD24/metabolismo , Decidua/metabolismo , Endometrio/metabolismoRESUMEN
INTRODUCTION: Abnormal placental trophoblast function is the main cause of missed abortion (MA). Src kinase-associated phosphoprotein 2 (SKAP2) indirectly affects actin reunion, which is significantly associated with cell migration. METHODS: Twenty women with MA and 20 healthy women who underwent voluntarily induced abortion were included in this study. Immunohistochemistry, qRT-PCR, and western blotting were used to determine SKAP2, WAVE2, and ARP2 expression in the villous tissues. We investigated the effects of SKAP2 and the W336K mutant (blocked SKAP2 Src homology 3 function) on growth and migration in HTR8/SVneo cells using the CCK8 assay, flow cytometry, and transwell assay. The effects of SKAP2 on the WAVE2-ARP2/3 signaling pathway in HTR8/SVneo cells were evaluated by western blotting and immunofluorescence. RESULTS: Compared to the women in the voluntary abortion group, SKAP2 and WAVE2 expression levels were downregulated in the villous of patients with MA. In HTR8/SVneo cells, SKAP2 siRNA silencing regulated the growth and migration, while SKAP2 overexpression promoted growth and migration, and inhibited apoptosis. Additionally, SKAP2 regulated the expression of WAVE2 and ARP2, as well as the colocalization of actin with WAVE2. The SKAP2 W336K mutant could not alter WAVE2 and ARP2 expression, nor HTR8/SVneo cell growth and migration, with or without SKAP2 siRNA transfection. DISCUSSION: SKAP2 could activate the WAVE2-ARP2/3 pathway resulting in an increase of growth and migration in trophoblasts. SKAP2 probably played an important role in MA by affecting the growth and migration of trophoblasts.
Asunto(s)
Aborto Retenido , Trofoblastos , Aborto Retenido/metabolismo , Actinas/metabolismo , Movimiento Celular , Femenino , Humanos , Péptidos y Proteínas de Señalización Intracelular , Fosfoproteínas/metabolismo , Placenta/metabolismo , Embarazo , ARN Interferente Pequeño/genética , Transducción de Señal , Trofoblastos/metabolismo , Familia-src Quinasas/metabolismo , Familia-src Quinasas/farmacologíaRESUMEN
Missed abortion (MA) is a special form of spontaneous abortion that is increasing in incidence. However, the precise molecular mechanisms underlying MA, especially regarding the decidua, are poorly understood. Herein, we identified molecular signaling pathways related to MA by comparing the decidua of women experiencing normal pregnancy and MA using a quantitative proteomics approach based on HPLC-MS/MS and iTRAQ labeling. Integrated bioinformatics analysis of villi and decidua was performed to reveal potential crosstalk signals in closely related tissues. We identified 2277 proteins with high confidence in decidua, of which 232 were differentially expressed in MA samples. Specifically, we reported that integrated quantitative proteomic and bioinformatic analysis revealed altered proteins in MA and the mechanisms underpinning MA involved numerous pathways, especially ribosome and cellular metabolism signaling. Moreover, Importin 9, Cullin 1 and COX6C are critical for MA, and their altered expression might contribute to the pathophysiology of MA. In particular, COX6C was dramatically down-regulated in both decidua and villi of MA. COX6C was also found to be highly expressed in syncytiotrophoblastic and cytotrophoblastic cells in villi and widely expressed in decidua of the control group, but dramatically decreased in the MA group. Functional analysis showed that knockdown of COX6C inhibited apoptosis process in both HTR-8 and SiHa cells, suggesting that COX6C may play protective effects in MA. Thus, this study could help to map the regulatory protein network related to MA and contribute to the pathophysiological mechanisms of MA.
Asunto(s)
Aborto Retenido , Aborto Retenido/metabolismo , Vellosidades Coriónicas/metabolismo , Decidua/metabolismo , Femenino , Humanos , Embarazo , Proteómica , Espectrometría de Masas en TándemRESUMEN
Missed abortion, one of the leading causes of maternal and perinatal mortality, is associated with impaired trophoblast function. Opa interacting protein 5 (OIP5) interacts with outer membrane proteins, Opa, to play an important role in mitosis and tumorigenesis. The role of OIP5 in missed abortion was investigated in this study. Firstly, the expression of OIP5 in villous samples from patients with missed abortion was compared with women with normal pregnancies. Result showed that OIP5 was down-regulated in the placental villi from patients with missed abortion. Secondly, human first-trimester extravillous trophoblast-derived cell line (HTR-8/SVneo) was transfected with shRNA targeting OIP5 (shOIP5) or pcDNA-OIP5 (OIP5). Data from MTT and flow cytometry assays demonstrated that knockdown of OIP5 reduced number of viable cells in HTR-8/SVneo, and promoted the cell apoptosis. However, over-expression of OIP5 increased the number of viable cells and suppressed the cell apoptosis in HTR-8/SVneo. Moreover, cell migration of HTR-8/SVneo was inhibited by silencing of OIP5, and OIP5 over-expression enhanced protein expression of matrix metallopeptidase (MMP) 2/9. Lastly, OIP5 contributed to phosphorylation of STAT3 (signal transducer and activator of transcription 3) in HTR-8/SVneo. Inhibition of STAT3 attenuated OIP5 over-expression-induced increase in number of viable cells and migration in HTR-8/SVneo. In conclusion, OIP5 contributed to the proliferation and migration of trophoblast cell through activation of STAT3 signaling.
Asunto(s)
Aborto Retenido , Trofoblastos , Aborto Retenido/metabolismo , Movimiento Celular , Proliferación Celular , Femenino , Humanos , Placenta/metabolismo , Embarazo , Factor de Transcripción STAT3/genética , Factor de Transcripción STAT3/metabolismo , Trofoblastos/metabolismoRESUMEN
OBJECTIVE: To analyze the effects of the Human Chorionic Gonadotropin beta (ß-hCG) and the VEGF-MEK/ERK signaling pathway on villi angiogenesis in early missed abortion. METHODS: A total of 12 cases of women with missed abortion and 12 cases of women who had induced abortion voluntarily without any disease were included in the present study. The age, pregnancy time and gestation period in the control group corresponded to the missed abortion group. Wes Simple Western system and qRT-PCR were used to detect the expression of VEGF-MEK/ERK signaling pathway related proteins and genes in villous. Radioimmunoassay and Enzyme-linked immunosorbent assay were used to detect ß-hCG and VEGF levels in serum. The microvascular density (MVD) in villous tissue was analyzed by immunohistochemical staining. RESULTS: The levels of ß-hCG and VEGF in serum, the expression of VEGF-MEK/ERK signaling pathway and MVD in villous tissue of the missed abortion group were lower than those of the control group. In addition, compared with the control group, the layers of trophoblasts of the villous tissue in the missed abortion group became thinner significantly, the number of cells reduced, the cell structures were disorganized, and parts of the trophoblast cells were absent. Correlational analysis showed that the protein expression of ERK1/2 was positively correlated with MVD in missed abortion group. CONCLUSIONS: Our results reveal that decreased production of ß-hCG in early pregnant women could down-regulate the expression of VEGF-MEK/ERK signal pathway, then reduce angiogenesis and eventually leading to the abnormal angiogenesis of villous, which may be an important mechanism of missed abortion.
Asunto(s)
Aborto Retenido/genética , Gonadotropina Coriónica Humana de Subunidad beta/fisiología , Vellosidades Coriónicas/irrigación sanguínea , Sistema de Señalización de MAP Quinasas/fisiología , Factor A de Crecimiento Endotelial Vascular/fisiología , Aborto Inducido/efectos adversos , Aborto Retenido/metabolismo , Aborto Espontáneo/genética , Aborto Espontáneo/metabolismo , Aborto Espontáneo/patología , Adulto , Estudios de Casos y Controles , Vellosidades Coriónicas/metabolismo , Vellosidades Coriónicas/patología , Femenino , Humanos , Neovascularización Patológica/genética , Neovascularización Patológica/metabolismo , Embarazo , Primer Trimestre del Embarazo/genética , Primer Trimestre del Embarazo/metabolismo , Adulto JovenRESUMEN
OBJECTIVE: Human leukocyte antigen-G (HLA-G) is a molecule that was first known to confer protection to the fetus from destruction by the immune system of its mother. HLA-G expression is mainly restricted to the fetal-maternal interface on the extravillous cytotrophoblast, placenta, amnion.Methods: The purpose of this study is to investigate the HLA-G and KIR2DL4 expression in chorionic villous among 2 groups with missed abortion: group 1 - 27cases with normal karyotype and group 2 - 22 with fetal polyploidy. Criteria of inclusion: abortive material from two groups of women with missed abortion; 6-12 weeks gestational age, singleton pregnancy, cytogenetic of chorionic villous was obligatory - normal fetal karyotype and polyploidy of fetus.Results: During IHC investigations the average relative area of HLA-G expression in trophoblast was counted (in 1st group 33.9 ± 3.5 and in 2nd group 38.6 ± 2.8). Expression of HLA-G the most verified in extravillous chorion stroma. The average relative area of KIR2DL4 receptor was not statistically different among two groups (31.6 ± 2.4 and 32.2 ± 1.7). CONCLUSIONS: These results suggest the role of HLA-G for the progression in early reproductive losses. Low expression of HLA-G is associated with pregnancy complications and can be one of the reasons for spontaneous abortion.
Asunto(s)
Aborto Retenido/metabolismo , Vellosidades Coriónicas/metabolismo , Antígenos HLA-G/metabolismo , Receptores KIR2DL4/metabolismo , Aborto Habitual/metabolismo , Aborto Habitual/patología , Aborto Retenido/patología , Adulto , Vellosidades Coriónicas/patología , Femenino , Edad Gestacional , Humanos , Inmunohistoquímica , Placenta/metabolismo , Placenta/patología , Embarazo , Primer Trimestre del Embarazo/metabolismo , Trofoblastos/metabolismo , Trofoblastos/patologíaRESUMEN
Early non-chromosome-related missed abortion (MA) is commonly associated with an altered immunological environment during pregnancy. Human decidual natural killer (dNK) cells, the most abundant lymphocyte population within the first-trimester maternal-fetal interface, are vital maternal regulators of immune tolerance mediating successful embryo implantation and placentation. Previous studies have shown that dNK cells may play a role in MA. However, the gene expression status and specific altered manifestations of dNK cells in patients with early MA remain largely unknown. Here, we show that MA dNK cells have distinct mRNA and lncRNA expression profiles through RNA sequencing, with a total of 276 mRNAs and 67 lncRNAs being differentially expressed compared with controls. Protein-protein interaction analysis of differentially expressed mRNAs was performed to identify hub genes and key modules. An lncRNA-mRNA regulatory network characterized by the small-world property was constructed to reveal the regulation of mRNA transcription by differential hub lncRNAs. Functional annotation of differentially expressed mRNAs and lncRNAs was performed to disclose their potential roles in MA pathogenesis. Our data highlight several enriched biological processes (immune response, inflammatory response, cell adhesion, and extracellular matrix [ECM] organization) and signaling pathways (cytokine-cytokine receptor interaction, ECM-receptor interaction, Toll-like receptor signaling pathway, and phosphatidylinositol signaling system) that may influence MA. This study is the first to demonstrate the involvement of altered mRNA and lncRNA expression profiles in the dNK cell pathogenesis of early MA, facilitating a better understanding of the underlying molecular mechanisms and the development of novel MA therapeutic strategies targeting key mRNAs and lncRNAs.
Asunto(s)
Aborto Retenido/patología , Decidua/patología , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Células Asesinas Naturales/patología , ARN Largo no Codificante/genética , ARN Mensajero/metabolismo , Aborto Retenido/genética , Aborto Retenido/metabolismo , Adulto , Decidua/metabolismo , Femenino , Perfilación de la Expresión Génica , Humanos , Células Asesinas Naturales/metabolismo , MicroARNs/genética , Embarazo , Mapas de Interacción de Proteínas , ARN Mensajero/genética , Transducción de Señal , TranscriptomaRESUMEN
AIMS: Previous evidence has demonstrated that oxidative stress is related to the pathogenesis of missed abortion (MA), but the specific mechanism remains obscure. The adaptor protein APPL1 is one of the differential proteins in chorionic trophoblasts. Thus, this study aimed to assess the potential influence of APPL1 on oxidative stress responses as well the possible molecular mechanisms involving in MA. MAIN METHODS: In the present study, the chorionic trophoblasts and the HTR-8/SVneo cell line were researched in vitro. Small interfering RNA (siRNA) was used to suppress the expression of APPL1. The fluorescent probes DHE and DCFH-DA were used to assess the intracellular reactive oxidative species (ROS). The activity of superoxide dismutase (SOD) was determined. Apoptosis was detected by TUNEL and flow cytometry. Cell viability was determined using Cell Counting Kit-8. Protein expression was detected by immunohistochemistry, western blotting, and reverse transcription-quantitative PCR. KEY FINDINGS: The application of oxidant in normal chorionic trophoblasts induced cell death and overproduction of ROS, which was consistent with MA. In addition, knockdown of APPL1 in HTR-8/SVneo cells resulted in increased ROS and apoptosis, which could be rescued by pretreatment with antioxidants. Mechanistically, we report that overproduction of ROS in trophoblasts and disturbed SOD, APPL1 and Nrf2/HO-1 antioxidant responses constitute important contributors to apoptosis. SIGNIFICANCE: Our results suggest that APPL1 has antioxidant properties that suppress oxidative stress and apoptosis via the Nrf2/HO-1 pathway. Moreover, antioxidant N-acetylcysteine (NAC) effectively restored the impaired antioxidative defense system elicited by excess ROS, as a potential therapeutic reagent for MA.
Asunto(s)
Aborto Retenido/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Apoptosis/fisiología , Hemo-Oxigenasa 1/metabolismo , Factor 2 Relacionado con NF-E2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/fisiología , Proteínas Adaptadoras Transductoras de Señales/genética , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Femenino , Técnicas de Silenciamiento del Gen , Humanos , Embarazo , ARN Interferente Pequeño/farmacología , Superóxido Dismutasa/metabolismo , Trofoblastos/metabolismoRESUMEN
The endometrium is a highly complex tissue that is vulnerable to subtle gene expression changes and is the first point of contact for an implanting blastocyst. Talin1 has previously been identified to regulate cytoskeleton and cell motility, however it has not been investigated in association with infertility. Herein, we presented that Talin1 dysregulation in the missed abortion endometrium would negatively influence endometrial adhesive capacity. Mechanistically, intracellular Talin1 inhibited the nuclear transportation of LIM and SH3 protein 1 (LASP1) and restored the expression of adhesion-associated protein. Moreover, extracellular Talin1 enforces endometrial epithelial cell adhesive capacity by interacting with Vitronectin (VTN) and activating the FAK/Src/ERK signalling pathway. This finding provides a novel insight into the potential use of Talin1 for managing endometrial epithelia cell adhesion. This study represents the first demonstration of Talin1 function in endometrial epithelial cell adhesion and endometrial receptivity. Our findings indicate that re-expression of Talin1 might represent a useful strategy for preventing and treating early pregnancy failure and infertility.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Adhesión Celular/fisiología , Proteínas del Citoesqueleto/metabolismo , Endometrio/citología , Células Epiteliales/metabolismo , Proteínas con Dominio LIM/metabolismo , Talina/metabolismo , Vitronectina/metabolismo , Aborto Retenido/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Adulto , Proteínas del Citoesqueleto/genética , Regulación hacia Abajo , Femenino , Regulación de la Expresión Génica/fisiología , Humanos , Proteínas con Dominio LIM/genética , Embarazo , Talina/genética , Vitronectina/genéticaRESUMEN
Missed abortion (MA) is a common disease in obstetrics and gynecology. More and more studies have focused on the relationship between miRNAs and pregnancy maintenance and its related diseases. The aim of this article is to explore the relationship between miRNA and MA. The expression of miR-98 were detected by in situ hybridization and real-time PCR. Cell proliferation, activity and migration were measured via Edu, MTT, and transwell assays. The target genes of miR-98 are identified by dual-luciferase activity assay. And the expression levels of target genes were determined by Western blot, real-time PCR and immunohistochemistry. miR-98 was significantly up-regulated in placental villi from over 35 years old MA patients compared with the age-matched normal pregnant women. Up-regulation of miR-98 suppressed the proliferation, activity and migration of the human trophoblast HTR-8/SVneo cell in vitro. miR-98 could bind to GDF6 and FAPP2 mRNA 3'-UTR and negatively regulate their expression. The downregulation of miR-98 promoted cell proliferation, then knockdown of GDF6 or FAPP2 inhibited miR-98-mediated cell proliferation. GDF6 and FAPP2 expression in the placental villi from MA patients were decreased compared to normal placental tissues. The expression of miR-98 in MA had an opposite relationship with the expression of GDF6 and FAPP2. Overexpression of miR-98 is associated with the occurrence of MA. miR-98 prevents proliferation, viability and migration of trophoblast cells partially through targeting GDF6 and FAPP2.
Asunto(s)
Aborto Retenido/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Movimiento Celular/fisiología , Proliferación Celular/fisiología , Factor 6 de Diferenciación de Crecimiento/metabolismo , MicroARNs/metabolismo , Trofoblastos/metabolismo , Aborto Retenido/genética , Proteínas Adaptadoras Transductoras de Señales/genética , Adulto , Línea Celular , Femenino , Factor 6 de Diferenciación de Crecimiento/genética , Humanos , MicroARNs/genética , Placenta/metabolismo , Embarazo , Regulación hacia ArribaRESUMEN
The contemporary world despite its enough developed medicine and generally highly enlightened population faces a great problem of vitamin, micro-element and nutrient deficiency turning to become the XXI century pandemic. Along with that significant growth of interest can be seen towards vitamin D importance for reproductive physiology. The fact is that vitamin D receptors (VDR) have been detected in women's ovarium tissue, fallopian tubes, decidua and placenta. Some recent years studies have proven that vitamin D may act as immune regulator during implantation. During early pregnancy the trophoblast release vitamin D, which produces anti-inflammatory reaction and also induce decidual tissue growth for successive pregnancy. It was a comparison between the expression of Vitamin D and VDR in chorionic villous in cases of normal pregnancy and missed abortion groups. 64 samples of chorionic villous were taken: 32 from missed abortion and 32 from the induced abortion group. Abortive material was taken from two groups of women residing in North-West region of Russia: missed abortion and pregnancy terminated at woman's wish (induced abortion); 6-12 weeks of gestation, singleton pregnancy. Immune histochemical examination showed homogenous distribution of vitamin D and VDR expression in syncytiotrophoblasts, cytotrophoblasts and chorion villus stroma.Vitamin D expression relative area was 10,3% which is statistically different from the induced abortion group - 15,4% (p<0,01). VDR expression analysis showed its homogenous distribution in chorionic villus structures in both groups. High VDR expression was detected in chorion villus stromal components. In missed abortion group, the morphometry results showed distinctly lower relative area of vitamin D expression against the comparison group (35,9 ± 1,8; 56,1 ± 2,4 p < 0,01). Also in missed abortion group, positively significant correlation has been determined between the level of vitamin D in blood and VDR relative area expression (r = 0,412). In missed abortion group, definite vitamin D and VDR expression decrease was detected compared to the induced abortion group. The results witness vitamin D importance for pregnancy progress.
Asunto(s)
Aborto Retenido/metabolismo , Vellosidades Coriónicas/metabolismo , Receptores de Calcitriol/metabolismo , Vitamina D/metabolismo , Aborto Inducido , Aborto Retenido/sangre , Adulto , Estudios de Casos y Controles , Corion/metabolismo , Corion/patología , Vellosidades Coriónicas/patología , Femenino , Humanos , Inmunohistoquímica , Placenta/metabolismo , Placenta/patología , Embarazo , Primer Trimestre del Embarazo/metabolismo , Trofoblastos/metabolismo , Trofoblastos/patología , Vitamina D/sangre , Deficiencia de Vitamina D/sangre , Deficiencia de Vitamina D/complicaciones , Deficiencia de Vitamina D/metabolismoRESUMEN
We hypothesize that progesterone causes tolerogenic maturation of myeloid dendritic cells (DCs) in human decidua of threatened miscarriage or missed abortion characterized by a distinct phenotype and cytokine production, including reduction of the main NK cell proliferation and cytotoxic factor interleukin (IL)-15. During DC/NK cell interaction, progesterone-shaped DCs cannot efficiently multiply or equip NK cells with the cytotoxic mediators peforin and granulysin, which might harm trophoblasts and induce abortion. We propose that the presence, and maturation stage of decidual myeloid DCs be investigated using semi-quantitative immunohistological analyses and/or double-color immuno-fluorescent labeling of DC lineage and activation markers. The spatial arrangement of granulysin+â¯cells, NKp46+â¯NK cells, DCs, and trophoblasts might provide information about their mutual interactions in vivo. Multiple flow cytometry analyses of NK-receptors would provide insight into NK cell activation status. NK cell activation status could be also assessed by cytotoxicity assays against trophoblast cell lines, or isolated cognate extra-villous trophoblast cells. A correlation between decidual progesterone concentration or IL-15 expression, and the degree of DC maturation or the frequency of granulysin+â¯cells, might help to elucidate the mechanism of abortion retention in utero.
Asunto(s)
Aborto Retenido/metabolismo , Células Dendríticas/citología , Fertilización , Progesterona/metabolismo , Comunicación Celular , Proliferación Celular , Citocinas/metabolismo , Decidua/citología , Femenino , Citometría de Flujo , Humanos , Inmunohistoquímica , Interleucina-15/metabolismo , Células Asesinas Naturales/citología , Activación de Linfocitos , Modelos Teóricos , Células Mieloides/citología , Fenotipo , Embarazo , Trofoblastos/citologíaRESUMEN
Missed abortion is a special form of spontaneous abortion and its incidence shows a rising trend. Immunological factor is one of the most common reasons. Tumor suppressor gene programmed cell death 4 ( PDCD4) also participates in some immune-mediated inflammation, such as atherosclerosis, and so on, but the role of PDCD4 in missed abortion remains unclear. Here, the expression of PDCD4 was detected in decidual and chorionic tissues, as well as peripheral blood mononuclear cells from patients with missed abortion and healthy controls using quantitative real-time polymerase chain reaction (qRT-PCR), Western blot, and immunohistochemistry. The expression of cytokines was also detected in decidual tissues using qRT-PCR. The levels of serum estradiol and progesterone were measured by radioimmunoassay. In addition, the correlations of PDCD4 expression with cytokines and hormones were analyzed. The results demonstrated that PDCD4 expression was reduced in decidual tissues from the missed abortion group compared with the control group. The levels of tumor necrosis factor α were significantly higher in decidual tissues of missed abortion patients than those in normal controls. We also found serum estradiol and progesterone levels were significantly lower in the missed abortion group than those in the control group, and serum progesterone level was inversely related to PDCD4 messenger RNA level. The data suggested that reduced PDCD4 expression may be involved in the occurrence of missed abortion. This may facilitate the potential development of novel diagnostic and therapeutic strategies for the treatment of missed abortion.
Asunto(s)
Aborto Retenido/genética , Aborto Retenido/metabolismo , Proteínas Reguladoras de la Apoptosis/biosíntesis , Proteínas Reguladoras de la Apoptosis/genética , Regulación del Desarrollo de la Expresión Génica , Proteínas de Unión al ARN/biosíntesis , Proteínas de Unión al ARN/genética , Aborto Retenido/patología , Adulto , Biomarcadores/metabolismo , Corion/metabolismo , Corion/patología , Decidua/metabolismo , Decidua/patología , Femenino , Humanos , Leucocitos Mononucleares/metabolismo , Leucocitos Mononucleares/patología , Embarazo , Progesterona/sangreRESUMEN
Objective: To compare the distribution of proopiomelanocortin (POMC) in decidua and placenta samples from missed abortion and voluntary termination cases in order to research the effects in the etiology of missed abortion. Study Design: Decidual materials were collected from patients who were diagnosed with missed abortion (n=19) and legal voluntary termination cases (n=15) under 10 gestational weeks. Materials were divided into 2 groups for examination. For all samples, POMC primary antibody was performed by immunohistochemical staining. The number of stained cells was calculated by using the H-score technique. Results: In the missed abortion group the mean age was 28.7 (1841), and in the control group the mean age was 27.5 (2137). POMC immunoreactivity was determined to be lower in the parenchyma and placenta of the missed abortion group than those of the control group. POMC immunoreactivities were found to be higher in both the syncytiotrophoblast and cytotrophoblast cells of the missed abortion group than those of the control group (p<0.005). Conclusion: POMC has become a paradigmatic polypeptide precursor and has a role in the parturition process. Local production of POMC in placenta and decidua may influence pregnancy and may have a role in missed abortion pathogenesis.
Asunto(s)
Aborto Retenido/metabolismo , Aborto Espontáneo/metabolismo , Decidua/metabolismo , Proopiomelanocortina/metabolismo , Aborto Espontáneo/prevención & control , Adulto , Estudios de Casos y Controles , Femenino , Humanos , Inmunohistoquímica , Neovascularización Patológica/metabolismo , Placenta/patología , Embarazo , Primer Trimestre del Embarazo/metabolismo , Trofoblastos/patología , Adulto JovenRESUMEN
OBJECTIVE: To investigate the function and underlying mechanism of transforming growth factorbeta (TGF-ß)/bone morphogenetic protein (BMP) signaling pathway in early unexplained miscarriage. STUDY DESIGN: Expression profiles of genes involved in TGF-ß/BMP signaling pathway were compared between placental villous tissue samples from 2 women with missed abortion and those from 2 women with induced abortion by microarray assay. The protein expression level of the most downregulated geneLEFTY1was further measured using western blotting in another 8 women with missed abortion and 7 women with induced abortion. RESULTS: A total of 24 genes showed differential expression level between the 2 groups. Their functions were further investigated, of which 6 of 13 upregulated genes were TGF-ß responsive genes. The most reduced gene is LEFTY1, an antagonist of TGF-ß ligand. The protein expression level of LEFTY1 was confirmed to show the same trend as microarray using western blotting. CONCLUSION: A reduced expression of LEFTY1 in women with missed abortion was identified as com-pared with women with induced abortion, which may result in a dysregulation of TGF-ß signaling and may be the underlying mechanism of missed abortion.
Asunto(s)
Aborto Retenido/metabolismo , Vellosidades Coriónicas , Factores de Determinación Derecha-Izquierda , Adulto , Vellosidades Coriónicas/química , Vellosidades Coriónicas/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Humanos , Factores de Determinación Derecha-Izquierda/análisis , Factores de Determinación Derecha-Izquierda/genética , Factores de Determinación Derecha-Izquierda/metabolismo , EmbarazoRESUMEN
INTRODUCTION: The invasion of extravillous cytotrophoblasts (EVTs) into the maternal uterine decidua and vasculature is critical for human placenta development and pregnancy maintenance. The imprinted gene MEST/PEG1 has been implicated in trophoblast development; however, the role of MEST in EVT invasion and the accompanying early pregnancy complications are not fully understood. METHODS: Western blot, immunofluorescence and immunohistochemistry were used to detect MEST protein expression and localization by using antibodies recognize 2 reported isoforms. Specific small interference RNA (siRNA) targeting both of the MEST isoforms was applied to silence MEST expression in extravillous explants and HTR8/SVneo cells. Cell invasion and migration were assessed using the Matrigel invasion, Transwell migration assay and the xCELLigence system. Promoter DNA methylation was examined using bisulfite-sequencing polymerase chain reaction (BSP). RESULTS: MEST protein was highly expressed in EVTs in the first trimester placenta and in the invasive EVT cell lines HTR-8/Svneo and HPT-8. Weak MEST expression was found in cytotrophoblasts (CTBs) and the choriocarcinoma-derived CTB cell line JEG-3. The specific siRNA knockdown of MEST expression significantly reduced HTR-8/Svneo cell invasion and migration as well as extravillous explant outgrowth, which were associated with the downregulation of Twist, N-cadherin and Vimentin. Decreased MEST protein expression with isoform 2 promoter hypermethylation was observed in the placentas of missed abortions, suggesting a possible pathological mechanism of missed abortion. CONCLUSIONS: Suppressed expression of MEST was associated with its isoform 2 promoter hypermethylation ex vivo placenta tissues and in vitro cultured EVT cell lines. The present results provide a possible pathological mechanism of missed abortion.
Asunto(s)
Aborto Retenido/metabolismo , Proteínas/metabolismo , Trofoblastos/metabolismo , Secuencia de Bases , Cadherinas/metabolismo , Metilación de ADN , Femenino , Humanos , Embarazo , Primer Trimestre del Embarazo/metabolismo , Regiones Promotoras Genéticas , Vimentina/metabolismoRESUMEN
Background/Aim: Recurrent or habitual missed abortions (RMA) are defined as three or more consecutive abortions. In the first trimester of pregnancy habitual missed abortions occur in about 1% of population. The aim of this immunohistochemical study of decidua in RMA of unknown etiology was to identify subpopulations of decidual lymphocytes in recurrent miscarriages and compare the distribution of immunocompetent cells in artificial abortions and RMA. Methods. The study included 30 women with at least 2 consecutive miscarriages in the first trimester of pregnancy. Curettements of the third missed abortion were immunohistochemically analyzed. The control group consisted of 20 women without loaded reproductive anamnesis, with the abortion for social reasons. Criteria for exclusion from the study were diagnosed uterine anomalies, positive screening for thrombophilia and women who suffered from diabetes mellitus and disorders in the function of the thyroid gland. Immunophenotyping was performed by immuno-alkaline phosphatase (APAAP) using monoclonal antibodies: CD 30, CD 45 RO, CD 56 and CD 57, CD 68. Methods: The number of missed abortions (1,223) was on the average 9.7% of all deliveriies during the test period. Among them RMA were registered in 52 (4.2%) patients and in 30 (57%) the exact etiology of abortions was not determined. RMA was most common in the 25-34 years of age group. The largest number of RMA showed the ultrasound characteristics of missed abortion in 60% of cases and was in nulliparous patients (76.7%). The number of NK CD56 positive cells did not differ significantly between the types of abortion. In the decidual tissue, a number of NK CD57 positive cells was significantly higher in missed abortions compared to artificial interruptions (p < 0.01). In artificial termination of pregnancy there was an absolute predominance of CD45RO lymphocyte subpopulations, whereas in the RMA group there was slightly greater predominance of CD30 positive cells. The completed analysis showed a significantly higher number of CD68 positive macrophages in a decidual tissue of RMA pregnancy (p < 0.01). Results: The number and phenotypic structure of NK cells are significantly different in normal pregnancy decidua and in RMA. The NK cell dominance is present in the RMA group, in favor of CD56+ and CD 57 of subpopulations with increased CD30 of T lymphocyte subpopulations. Macrophages are more numerous in the decidua of pregnancies ended in abortion, so the cause to RMA of unknown etiology in a number of cases could be disregulation of immunocompetent cells.
Asunto(s)
Aborto Retenido/inmunología , Decidua/inmunología , Decidua/metabolismo , Células Asesinas Naturales/inmunología , Aborto Retenido/metabolismo , Adulto , Antígeno CD56 , Antígenos CD57 , Femenino , Humanos , Inmunohistoquímica , Inmunofenotipificación , Antígeno Ki-1 , Células Asesinas Naturales/clasificación , Células Asesinas Naturales/metabolismo , EmbarazoRESUMEN
BACKGROUND: Forkhead transcription factors 3a (FOXO3a) has pleiotropic biological functions in the female reproductive tract. FOXO3a has a function in decidualization, in placental development, and also in inhibition of apoptosis. This study aims to investigate a possible role of FOXO3a in missed abortion. MATERIALS AND METHODS: Decidual and placental tissue samples were obtained from the women with unwanted pregnancy as the control group and with missed abortion as the patient group. Immunohistochemistry technique was utilized to compare FOXO3a expression of the decidual cells in uterine decidual stroma and cytotrophoblast-syncytiotrophoblast cells in placental villous stroma. Immunohistochemistry was evaluated semiquantitatively utilizing the H-score technique. Results: It was demonstrated that H-Scores of FOXO3a expression in both uterine decidual stroma were increased in the missed abortion group (255.83 +/- 12.41) than in the normal pregnancy group (133.33 +/- 17.43). It was also shown that there was no difference between non-decidual area of the endometrium of the normal pregnancy and the missed abortion group (30.33 +/- 4.32; 39.66 +/- 14.30, respectively) and placental villous stroma (13.00 +/- 1.89; 13.00 +/- 1.67, respectively). However, the immunoreactivity of cytotrophoblast and syncytiotrophoblast cells significantly increased in the missed abortion group (18.83+1.47; 322.00 +/- 6.06, respectively) than in the normal pregnancy group (11.00 +/- 1.26; 254.00 +/- 8.17, respectively) (p < 0.05). CONCLUSION: These data support the hypothesis that increased FOXO3a expression in missed abortion may prevent the discharge of dead fetus to maintain decidualization, prevention of oxidative stress, immunomodulation, and inhibition of apoptosis.