Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 22
Filtrar
1.
Mutagenesis ; 38(1): 64-70, 2023 02 03.
Artigo em Inglês | MEDLINE | ID: mdl-36082793

RESUMO

An increase of reactive oxygen species in the placenta and oxidative disbalance has been recognized as a significant factor contributing to pregnancy complications. Dietary intake of food rich in antioxidants during pregnancy could exert a protective role in the prevention of adverse outcomes such as preeclampsia, miscarriage, and others. Flavonoid taxifolin has shown numerous health-promoting effects in a large number of studies conducted on animals, as well as various human cell types in vitro. However, its effects on human placental cells-trophoblasts-have yet to be determined. Therefore, cytoprotective and genoprotective effects of taxifolin on trophoblast cell line HTR-8/SVneo under induced oxidative stress were explored in this study. Cytotoxicity of a range of taxifolin concentrations (1-150 µM) was evaluated using the MTT and crystal violet assays. A model of oxidative stress was achieved by exposing HTR-8/SVneo cells to H2O2. To determine cytoprotective and antigenotoxic effects, the cells were pre-incubated with three concentrations of taxifolin (10, 50, and 100 µM) and then exposed to H2O2. Taxifolin in concentrations of 1, 5, 10, 25, 50, and 100 µM showed no cytotoxic effects on HTR-8/SVneo cells, but 150 µM of taxifolin caused a significant decrease in adherent cell number, as detected by crystal violet assay. Pretreatment with the chosen concentrations of taxifolin showed a significant cytoprotective effect on H2O2-induced cytotoxicity, as determined by the MTT assay. Furthermore, taxifolin showed a significant reduction in H2O2-induced DNA damage, measured by comet assay. This study showed protective effects of taxifolin on human trophoblast cells exposed to oxidative damage. Further studies are needed to explore the underlying mechanisms.


Assuntos
Placenta , Trofoblastos , Humanos , Gravidez , Feminino , Trofoblastos/metabolismo , Placenta/metabolismo , Peróxido de Hidrogênio/farmacologia , Violeta Genciana/metabolismo , Violeta Genciana/farmacologia , Linhagem Celular , Estresse Oxidativo
2.
Mutagenesis ; 38(3): 169-181, 2023 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-37228020

RESUMO

Dry olive leaf extract (DOLE) and its active component oleuropein (OLE) were applied as reducing and stabilizing agents to prepare colloidal 20-25 nm silver nanoparticles (Ag NPs). The Ag NPs were characterized using transmission electron microscopy, X-ray diffraction analysis, and absorption spectroscopy. The cytotoxic actions of coated Ag NPs, and their inorganic and organic components, were examined against trophoblast cells and human peripheral blood lymphocytes (PBLs), Gram-positive, Gram-negative bacteria, and yeast. The genotoxic potential was evaluated in PBLs in vitro with the comet assay. Ag/DOLE and Ag/OLE induced cytotoxic effects in both types of cells after 24 h exposure when silver concentrations were 0.025-0.2 mM. However, the most pronounced cytotoxicity exhibits Ag/OLE. Both colloids also caused reduced ROS production in both cell types at 0.1 mM and 0.2 mM, while bare Ag NPs did not alter ROS levels at any of the conditions. Functionalized Ag/DOLE and Ag/OLE did not show genotoxic effects in PBLs, while bare AgNPs increased DNA damage significantly only at 0.2 mM. Regarding the antimicrobial effects, the Ag/OLE had MIC values for all evaluated microorganisms from 0.0625 to less than 0.0312 mM. Also, the antimicrobial effect of Ag/DOLE was significantly higher on Gram-negative bacteria and yeast than on Gram-positive bacteria. Obtained results indicate that Ag/OLE induced the most pronounced biological effects, beneficial for its application as an antimicrobial agent, but with potential risks from exposure to high concentrations that could induce cytotoxicity in healthy human cells.


Assuntos
Anti-Infecciosos , Nanopartículas Metálicas , Humanos , Prata/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Nanopartículas Metálicas/toxicidade , Nanopartículas Metálicas/química , Saccharomyces cerevisiae/metabolismo , Trofoblastos/metabolismo , Anti-Infecciosos/toxicidade , Anti-Infecciosos/química , Linfócitos/metabolismo
3.
Int J Mol Sci ; 25(1)2023 Dec 29.
Artigo em Inglês | MEDLINE | ID: mdl-38203672

RESUMO

Successful pregnancy establishment requires highly synchronized cross talk between the invasive trophoblast cells and the receptive maternal endometrium. Any disturbances in this tightly regulated process may lead to pregnancy complications. Local factors such as nutrients, hormones, cytokines and reactive oxygen species modulate the invasion of extravillous trophoblasts through critical signaling cascades. Epidemiological studies strongly indicate that a Mediterranean diet can significantly impact molecular pathways during placentation. Therefore, the aim of the current study was to examine whether oleuropein (OLE), one of the main compounds of the Mediterranean diet, may influence trophoblast cell adhesion and migration, as well as the expression of invasion-associated molecular markers and inflammatory pathways fostering these processes. HTR-8/SVneo cells were incubated with OLE at selected concentrations of 10 and 100 µM for 24 h. Results showed that OLE did not affect trophoblast cell viability, proliferation and adhesion after 24 h in in vitro treatment. The mRNA expression of integrin subunits α1, α5 and ß1, as well as matrix-degrading enzymes MMP-2 and -9, was significantly increased after treatment with 10 µM OLE. Furthermore, OLE at a concentration of 10 µM significantly increased the protein expression of integrin subunits α1 and ß1. Also, OLE inhibited the activation of JNK and reduced the protein expression of COX-2. Finally, a lower concentration of OLE 10 µM significantly stimulated migration of HTR-8/SVneo cells. In conclusion, the obtained results demonstrate the effects of OLE on the function of trophoblast cells by promoting cell migration and stimulating the expression of invasion markers. As suggested from results, these effects may be mediated via inhibition of the JNK signaling pathway.


Assuntos
Glucosídeos Iridoides , Trofoblastos , Feminino , Gravidez , Humanos , Glucosídeos Iridoides/farmacologia , Trofoblastos Extravilosos , Integrinas
4.
Int J Mol Sci ; 23(23)2022 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-36498901

RESUMO

Interleukin-6 (IL-6) is an acknowledged inflammatory cytokine with a pleiotropic action, mediating innate and adaptive immunity and multiple physiological processes, including protective and regenerative ones. IL-8 is a pro-inflammatory CXC chemokine with a primary function in attracting and activating neutrophils, but also implicated in a variety of other cellular processes. These two ILs are abundantly expressed at the feto-maternal interface over the course of a pregnancy and have been shown to participate in numerous pregnancy-related events. In this review, we summarize the literature data regarding their role in healthy and pathological pregnancies. The general information related to IL-6 and IL-8 functions is followed by an overview of their overall expression in cycling endometrium and at the feto-maternal interface. Further, we provide an overview of their involvement in pregnancy establishment and parturition. Finally, the implication of IL-6 and IL-8 in pregnancy-associated pathological conditions, such as pregnancy loss, preeclampsia, gestational diabetes mellitus and infection/inflammation is discussed.


Assuntos
Interleucina-6 , Pré-Eclâmpsia , Gravidez , Feminino , Humanos , Interleucina-8/genética , Citocinas , Parto
5.
Int J Mol Sci ; 23(1)2021 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-35008499

RESUMO

Galectins are a family of conserved soluble proteins defined by an affinity for ß-galactoside structures present on various glycoconjugates. Over the past few decades, galectins have been recognized as important factors for successful implantation and maintenance of pregnancy. An increasing number of studies have demonstrated their involvement in trophoblast cell function and placental development. In addition, several lines of evidence suggest their important roles in feto-maternal immune tolerance regulation and angiogenesis. Changed or dysregulated galectin expression is also described in pregnancy-related disorders. Although the data regarding galectins' clinical relevance are still at an early stage, evidence suggests that some galectin family members are promising candidates for better understanding pregnancy-related pathologies, as well as predicting biomarkers. In this review, we aim to summarize current knowledge of galectins in early pregnancy as well as in pregnancy-related pathologies.


Assuntos
Galectinas/metabolismo , Complicações na Gravidez/metabolismo , Animais , Feminino , Humanos , Placenta/metabolismo , Placentação/fisiologia , Gravidez , Trofoblastos/metabolismo
6.
Reprod Fertil Dev ; 2020 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-33323165

RESUMO

Extravillous trophoblasts are specific placental cells that invade the uterine stroma and spiral arteries modifying and adjusting them to pregnancy. Many pregnancy pathologies are associated with impairment of this process, including preeclampsia and intrauterine growth restriction, among others. Macrophage migration inhibitory factor (MIF) is a pleiotropic cytokine that is abundant at the fetomaternal interface. Previous results from our group showed that MIF participates in trophoblast invasion and modulates the expression of molecules known to mediate stromal and endovascular trophoblast invasion. In this study we investigated the possibility that MIF could act as a regulator of cytokines known to modulate trophoblast invasion using the normal extravillous trophoblast-derived cell line HTR-8/SVneo. Expression of trophoblast MIF was attenuated by MIF mRNA-specific small interfering RNAs. Cytokine expression was assessed at the mRNA and protein levels using real-time quantitative polymerase chain reaction and flow cytometry respectively. Knockdown of MIF led to a significant decrease in mRNA for IL-1ß (IL1B) and IL-8 (CXCL8) and interleukin (IL)-8 protein. The addition of recombinant human MIF to cell culture medium increased IL-6 after 24h treatment and IL-6 and IL-8 after 72h treatment. Cell viability was not affected by MIF silencing or rhMIF treatment. The results of this study imply that at least some of the effects of MIF on trophoblast invasion could be mediated through autocrine or paracrine modulation of trophoblast cytokine production.

7.
Histochem Cell Biol ; 142(5): 541-53, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24854997

RESUMO

Mucins are multifunctional highly glycosylated proteins expressed by the female reproductive tract. Differential expression of MUC1 and MUC15 has been shown in trophoblast. This study was undertaken to establish the distribution of mucin(s) in cytotrophoblast cell cultures using anti-bovine submaxillary mucin (BSM) and to investigate the possibility of MUC1/mucin(s) being a binding partner of trophoblast galectin-1. MUC1 is demonstrated here using immunocytochemistry on isolated cytotrophoblast and the HTR-8/SVneo extravillous trophoblast cell line but detection of additional trophoblast mucins cannot be excluded. Western blot analysis showed similar bands ranging from 30 to >200 kDa with anti-BSM and the well-known mucin antibodies HMFG1 and B72.3. Immunocytochemistry and cell-based ELISA data were found to support that all of the antibodies used are reactive with BSM, suggesting the presence of shared epitopes between BSM and trophoblast mucin(s). Binding of galectin-1 to trophoblast MUC1/mucin(s) was analyzed using a solid-phase assay and co-immunoprecipitation. Recombinant galectin-1 binding to isolated trophoblast mucin in solid-phase assay was sensitive to lactose, a carbohydrate inhibitor of galectin binding. In whole HTR-8/SVneo lysates, ~200 kDa mucin was detected in galectin-1 immunoprecipitates, while endogenous galectin-1 was present in BSM-immunoprecipitates. Furthermore, double fluorescence immunocytochemistry showed overlap of galectin-1 and trophoblast mucins at the plasma membrane of HTR-8/SVneo cells. These results suggest that trophoblast mucin(s) could act as binding partners of galectin-1, in a carbohydrate-dependent manner.


Assuntos
Galectina 1/metabolismo , Mucinas/metabolismo , Trofoblastos/metabolismo , Células Cultivadas , Galectina 1/química , Humanos , Imuno-Histoquímica , Mucinas/química , Ligação Proteica , Trofoblastos/citologia
8.
Reprod Toxicol ; 126: 108585, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38574953

RESUMO

Oxidative stress has been implicated in numerous pregnancy-related disorders. Biologically active plant secondary metabolites, which are present in everyday diet, could prove effective therapeutic agents in preventing these disorders. This study evaluated effects of taxifolin (dihydroquercetin) on ROS production, markers of oxidative damage to lipids and proteins, activity of antioxidant enzymes and production of pro-inflammatory cytokines in H2O2-induced oxidative stress in trophoblast HTR-8/SVneo cells. Taxifolin in 10 µM and 100 µM concentrations attenuated oxidative damage to lipids and proteins, as evidenced by a decrease in MDA content, extracellular LDH activity, carbonyl groups and nitrite contents. A reduction in the activity of antioxidant enzymes SOD, CAT and GPx in cells pre-treated with taxifolin, prior to H2O2 exposure, was also observed, along with a reduction in intracellular ROS production. Both evaluated concentrations of taxifolin showed anti-inflammatory activity in trophoblast cells, by reducing production of pro-inflammatory cytokines IL-1ß and IL-6. In this model of H2O2-induced oxidative stress, taxifolin showed marked antioxidative and anti-inflammatory activities in trophoblast cells, adding further evidence of its protective effects and showing potential as a therapeutic agent in preventing adverse pregnancy outcomes.


Assuntos
Anti-Inflamatórios , Antioxidantes , Peróxido de Hidrogênio , Estresse Oxidativo , Quercetina , Espécies Reativas de Oxigênio , Trofoblastos , Quercetina/análogos & derivados , Quercetina/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Peróxido de Hidrogênio/toxicidade , Humanos , Trofoblastos/efeitos dos fármacos , Trofoblastos/metabolismo , Antioxidantes/farmacologia , Linhagem Celular , Anti-Inflamatórios/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Catalase/metabolismo
9.
Pharmaceutics ; 16(6)2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38931863

RESUMO

The aims of the present research include (1) optimization of extraction from Vaccinium myrtillus leaf waste via investigation of plant material:medium ratio, extraction medium, and extraction period, employing extractions at room and high temperatures, or using ultrasound and microwaves (M, HAE, UAE, and MAE, respectively), (2) physicochemical characterization, and (3) investigation of extract biological potential. The statistical analysis revealed that optimal levels of parameters for the greatest polyphenolic yield were a proportion of 1:30 g/mL, ethyl alcohol 50% (v/v) during 2 min of microwave irradiation. By LC-MS analysis, 29 phenolic components were detected; HAE showed the highest richness of almost all determined polyphenols, while chlorogenic acid and quercetin 3-O-glucuronide were dominant. All extracts showed a high inhibition of Staphylococcus aureus growth. The effect of different parameters on extracts' antioxidant capacity depended on the used tests. The extracts also showed a stimulative influence on keratinocyte viability and anti-inflammatory activity (proven in cell-based ELISA and erythrocyte stabilization assays). The extraction procedure significantly affected the extraction yield (MAE ≥ maceration ≥ UAE ≥ HAE), whereas conductivity, density, surface tension, and viscosity varied in a narrow range. The presented research provides evidence on the optimal extraction conditions and technique, chemical composition, and antioxidant, antimicrobial, anti-inflammatory, and keratinocyte viability properties of bilberry extracts for potential applications in pharmacy and cosmetics.

10.
Plants (Basel) ; 12(14)2023 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-37514358

RESUMO

In the study, the optimization of the extraction from Aloe vera leaf waste was performed via varying solid-to-solvent ratio, solvent type, extraction time, and technique (maceration, heat-, ultrasound-, and microwave-assisted extractions-HAE, UAE, and MAE, respectively). The optimal extraction conditions for achieving the highest polyphenol content are a 1:30 ratio, 70% ethanol, and 30 min of HAE. Total flavonoid and protein contents were significantly higher in the extract from MAE, while total condensed tannin content was the highest in HAE. LC-MS analysis quantified 13 anthraquinone and chromone compounds. The variations in the FT-IR spectra of the extracts obtained by different extraction procedures are minor. The influence of extraction conditions on the antioxidant ability of the extracts depended on applied antioxidant assays. The extracts possessed medium inhibition properties against Staphylococcus aureus and weak inhibitory activity against Enterococcus feacalis. The extracts had stimulative effect on HaCaT cell viability. Regarding the extraction yield, there was a significant difference between the used extraction techniques (MAE > HAE > maceration and UAE). The presented study is an initial step in the production of polyphenol-rich extracts from A. vera leaf waste aimed to be used for the potential preparation of pharmaceutical and cosmetic formulations for the skin.

11.
Biomedicines ; 11(10)2023 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-37893055

RESUMO

Antiphospholipid syndrome (APS) is a complex thrombo-inflammatory autoimmune disease characterized by the presence of antiphospholipid antibodies (aPL). Women with APS are at high risk of recurrent early pregnancy loss as well as late obstetrical complications-premature birth due to placental insufficiency or severe preeclampsia. Accumulating evidence implies that vascular thrombosis is not the only pathogenic mechanism in obstetric APS, and that the direct negative effect of aPL on the placental cells, trophoblast, plays a major role. In this review, we summarize the current findings regarding the potential mechanisms involved in aPL-induced trophoblast dysfunction. Introduction on the APS and aPL is followed by an overview of the effects of aPL on trophoblast-survival, cell function and aPL internalization. Finally, the implication of several non-coding RNAs in pathogenesis of obstetric APS is discussed, with special emphasis of their possible role in trophoblast dysfunction and the associated mechanisms.

12.
Nutrients ; 14(24)2022 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-36558404

RESUMO

Polyphenols are a group of phytochemicals with extensive biological functions and health-promoting potential. These compounds are present in most foods of plant origin and their increased widespread availability through the intake of nutritional supplements, fortified foods, and beverages, has also led to increased exposure throughout gestation. In this narrative review, we focus on the role of polyphenols in both healthy and pathological pregnancy. General information related to their classification and function is followed by an overview of their known effects in early-pregnancy events, including the current insights into molecular mechanisms involved. Further, we provide an overview of their involvement in some of the most common pregnancy-associated pathological conditions, such as preeclampsia and gestational diabetes mellitus. Additionally, we also discuss the estimated possible risk of polyphenol consumption on pregnancy outcomes. The consumption of dietary polyphenols during pregnancy needs particular attention considering the possible effects of polyphenols on the mechanisms involved in maternal adaptation and fetal development. Further studies are strongly needed to unravel the in vivo effects of polyphenol metabolites during pregnancy, as well as their role on advanced maternal age, prenatal nutrition, and metabolic risk of the offspring.


Assuntos
Suplementos Nutricionais , Polifenóis , Gravidez , Feminino , Humanos , Polifenóis/farmacologia , Fenômenos Fisiológicos da Nutrição Pré-Natal , Desenvolvimento Fetal , Alimentos Fortificados
13.
Food Chem Toxicol ; 163: 112993, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35398184

RESUMO

Caffeic acid is highlighted as one of the major phenolic compounds present in foods with known antioxidant activity. This phenolic is among commonly consumed substances in everyday diet of pregnant women. However, there is not enough information on its effects during pregnancy, especially the most vulnerable early stage. Extravillous trophoblast cells are specific cells of the placenta that come in direct contact with maternal uterine tissue. Through this study we investigated the cytoprotective effects of caffeic acid on H2O2-induced oxidative damage in first trimester extravillous trophoblast cell line HTR-8/SVneo. Investigated concentrations (1-100 µM) of caffeic acid showed neither cytotoxic nor genotoxic effects on HTR-8/SVneo cells. The treatment with caffeic acid 100 µM significantly increased the percentage of cells in G2/M phase of the cell cycle, compared to non-treated cells. Pretreatment with caffeic acid (10 and 100 µM) attenuated oxidative DNA damage significantly, reduced cytotoxicity, protein and lipid peroxidation, and restored antioxidant capacity in trophoblast cells following H2O2 exposure. This beneficial outcome is probably mediated by the augmentation of GSH and effective ROS scavenging by caffeic acid. These promising results require further investigations to reveal the additional mechanisms/pathways and confirmation through studies in vivo.


Assuntos
Peróxido de Hidrogênio , Trofoblastos , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Ácidos Cafeicos , Linhagem Celular , Movimento Celular , Dano ao DNA , Feminino , Humanos , Peróxido de Hidrogênio/metabolismo , Estresse Oxidativo , Placenta , Gravidez
14.
Nanomaterials (Basel) ; 12(18)2022 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-36144978

RESUMO

The antibacterial performance and cytotoxic examination of in situ prepared silver nanoparticles (Ag NPs), on inorganic-organic hybrid nanopowder consisting of zirconium dioxide nanoparticles (ZrO2 NPs) and dihydroquercetin (DHQ), was performed against Gram (-) bacteria Escherichia coli and Gram (+) bacteria Staphylococcus aureus, as well as against human cervical cancer cells HeLa and healthy MRC-5 human cells. The surface modification of ZrO2 NPs, synthesized by the sol-gel method, with DHQ leads to the interfacial charge transfer (ICT) complex formation indicated by the appearance of absorption in the visible spectral range. The prepared samples were thoroughly characterized (TEM, XRD, reflection spectroscopy), and, in addition, the spectroscopic observations are supported by the density functional theory (DFT) calculations using a cluster model. The concentration- and time-dependent antibacterial tests indicated a complete reduction of bacterial species, E. coli and S. aureus, for all investigated concentrations of silver (0.10, 0.25, and 0.50 mg/mL) after 24 h of contact. On the other side, the functionalized ZrO2 NPs with DHQ, before and after deposition of Ag NPs, do not display a significant decrease in the viability of HeLa MRC-5 cells in any of the used concentrations compared to the control.

15.
Placenta ; 116: 51-57, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-33612316

RESUMO

Macrophage migration inhibitory factor (MIF) is a versatile cytokine acting as an important regulator of innate and adaptive immunity and implicated in many physiological and pathological processes. It is abundantly expressed at the feto-maternal interface and proposed to have a role in establishing and maintaining a healthy pregnancy. This review presents the current literature data regarding the MIF role in early pregnancy events and its association with some of the placental pathological conditions, including infection, preeclampsia, gestational diabetes mellitus and choriocarcinoma. General information regarding MIF structure and function is followed by an overview of its expression in reproductive tissues and in pregnancy. Futher, we discuss MIF's involvement in the survival of decidual stromal cells, placenta of the first trimester of pregnancy, and in trophoblast cell functions studied in vitro. Current findings associating this cytokine to placental infection, preeclampsia, gestational diabetes mellitus and choriocarcinoma are presented in the final part.


Assuntos
Fatores Inibidores da Migração de Macrófagos/metabolismo , Doenças Placentárias/metabolismo , Placenta/metabolismo , Coriocarcinoma/metabolismo , Coriocarcinoma/patologia , Feminino , Humanos , Placenta/patologia , Doenças Placentárias/patologia , Gravidez , Trofoblastos/metabolismo , Trofoblastos/patologia , Neoplasias Uterinas/metabolismo , Neoplasias Uterinas/patologia
16.
Chem Biol Interact ; 347: 109618, 2021 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-34364836

RESUMO

The toxicity of hybrid nanoparticles, consisting of non-toxic components, zirconium dioxide nanoparticles (ZrO2 NPs), and caffeic acid (CA), was examined against four different cell lines (HTR-8 SV/Neo, JEG-3, JAR, and HeLa). Stable aqueous ZrO2 sol, synthesized by forced hydrolysis, consists of 3-4 nm in size primary particles organized in 30-60 nm in size snowflake-like particles, as determined by transmission electron microscopy and direct light scattering measurements. The surface modification of ZrO2 NPs with CA leads to the formation of an interfacial charge transfer (ICT) complex followed by the appearance of absorption in the visible spectral range. The spectroscopic observations are complemented with the density functional theory calculations using a cluster model. The ZrO2 NPs and CA are non-toxic against four different cell lines in investigated concentration range. Also, ZrO2 NPs promote the proliferation of HTR-8 SV/Neo, JAR, and HeLa cells. On the other hand, hybrid ZrO2/CA NPs induced a significant reduction of the viability of the JEG-3 cells (39 %) for the high concentration of components (1.6 mM ZrO2 and 0.4 mM CA).


Assuntos
Ácidos Cafeicos/toxicidade , Nanopartículas Metálicas/toxicidade , Placenta/efeitos dos fármacos , Zircônio/toxicidade , Ácidos Cafeicos/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Teoria da Densidade Funcional , Feminino , Humanos , Nanopartículas Metálicas/química , Modelos Químicos , Tamanho da Partícula , Placenta/patologia , Gravidez , Testes de Toxicidade , Zircônio/química
17.
EXCLI J ; 18: Doc1007, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31762725

RESUMO

Macrophage migration inhibitory factor (MIF) is a multifunctional cytokine abundantly present at the feto-maternal interface proposed to play a role in establishment of pregnancy. We have previously shown that pharmacological inhibition of enzymatic activity of MIF decreases extravillous trophoblast invasion and migration in vitro. This study aimed to further elucidate potential role of endogenous trophoblast MIF, and to assess its importance for endovascular trophoblast cell function in particular. Attenuation of MIF by siRNA reduced HTR-8/SVneo cell invasion through Matrigel (59 % of control), expression of integrin α1 (86 % of control) and levels of MMP2 and MMP9 (87 % and 57 % of control, respectively). MIF specific siRNA reduced the ability of HTR-8/SVneo to differentiate in to endothelial-like phenotype, as determined by Matrigel tube formation assay. The total tube length was decreased to 68.6 %, while the number of branching points was reduced to 57.8 % of control. HTR-8/SVneo cell capacity to integrate into HUVEC monolayers was reduced by knock-down of MIF. This could be partly caused by reduced N-cadherin expression to 63 % of control, which decreased with knock-down of MIF, as the expression of this protein was recently shown essential for trophoblast-endothelial interaction. These novel findings indicate a novel role for trophoblast MIF in spiral artery remodeling process.

18.
Immunobiology ; 223(10): 544-548, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29960807

RESUMO

Women with antiphospholipid syndrome (APS) experience pregnancy complications mostly due to impaired trophoblast cell functions. Antiphospholipid antibodies (aPL) affect extravillous trophoblast in vivo and in culture, but the mechanisms are still poorly understood. Previously, syncytiotrophoblast was shown to bind and internalize aPL, which was not replicated for extravillous cytotrophoblast in short term culture. Here, aPL binding and time dependent internalization was demonstrated with exposure to aPL in the extravillous cell line HTR-8/SVneo and isolated first trimester of pregnancy cytotrophoblast (CT) using immunocytochemistry and flow cytometry. Intracellular aPL were detectable from 2 h of culture, reaching 30.7 ±â€¯3.1% (p < 0.001) positive cells in CT and 24.8 ±â€¯7% (p < 0.01) in HTR-8/SVneo cells at 24 h and 33 ±â€¯4.2% (p < 0.01) at 48 h. The data presented show that extravillous trophoblast cells internalize aPL in a time-dependent manner significantly more than control immunoglobulins after 24 h of exposure.


Assuntos
Anticorpos Antifosfolipídeos/sangue , Síndrome Antifosfolipídica/imunologia , Complicações na Gravidez/imunologia , Primeiro Trimestre da Gravidez/imunologia , Trofoblastos/imunologia , Síndrome Antifosfolipídica/sangue , Linhagem Celular , Vilosidades Coriônicas/imunologia , Feminino , Humanos , Gravidez , Complicações na Gravidez/sangue
19.
J Biochem ; 163(1): 39-50, 2018 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-28992109

RESUMO

Interaction of sugar binding proteins-galectins, with glycoconjugates is considered relevant for various reproductive processes. Galectin-1 (gal-1) is a molecule involved in trophoblast cell invasion, which is accomplished through interaction with cell surface and/or extracellular matrix glycoproteins. A possibility of interaction of endogenous gal-1 and trophoblast ß1 integrins, both previously shown relevant for trophoblast invasion, was investigated. Confocal microscopy showed overlap in gal-1 and ß1 integrin localization at the plasma membrane of isolated cytotrophoblast, HTR-8/SVneo extravillous trophoblast cell line and JAr choriocarcinoma cells. Immunoprecipitation confirmed an interaction of gal-1 with integrin ß1, but not with α1 or α5 integrin subunits. Nondenaturing electrophoresis and subcellular fractionation suggested association of gal-1 with ß1 integrin in intracellular and plasma membrane compartments of HTR-8/SVneo cells. Gal-1/ß1 integrin complex was sensitive to chemical and enzyme treatments, indicating carbohydrate dependent interaction. Down-regulation of gal-1 by siRNA, however, had no effect on level or distribution of ß1 integrin, as determined by qPCR and flow cytometry. These results suggest complex lectin type interaction of gal-1 with ß1 integrin at the trophoblast cell membrane, which could influence trophoblast cell adhesion, migration and invasion.


Assuntos
Galectina 1/metabolismo , Integrina beta1/metabolismo , Trofoblastos/metabolismo , Células Cultivadas , Galectina 1/química , Galectina 1/genética , Humanos , Integrina beta1/química , Modelos Moleculares , Trofoblastos/citologia
20.
J Matern Fetal Neonatal Med ; 30(18): 2232-2236, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27690725

RESUMO

PURPOSE: Gal-3, which can regulate immune responses upon infection and inflammation, was not studied so far in intrauterine infection leading to preterm prelabor rupture of the membranes (PPROM), although gal-1 was reported to be implicated in the process. Gal-3 mRNA and protein expression in amnion and its changes during histological chorioamnionitis were studied here. MATERIALS AND METHODS: Fetal membranes were obtained from women with PPROM with (n =15) and without histological chorioamnionitis (n =15) during second and third trimester. Immunohistochemical reactivity was evaluated semiquantitatively and analyzed using t-test. Galectin profile of amniotic epithelia was determined by polymerase chain reaction (PCR) and change assessed in gal-3 in PPROM with (n =5) or without histological chorioamnionitis (n =5) by real-time PCR. RESULTS: Human amniotic epithelium was found to express gal-1, gal-3, gal-7 and gal-8 mRNA. Gal-3 mRNA and protein is increased in fetal membranes and in the amniotic epithelium in patients with chorionamnionitis. CONCLUSION: Histological chorioamnionitis is associated with increased gal-3 expression and strong immunoreactivity of the amnion. Gal-3 may participate in the regulation of the inflammatory responses to chorioamniotic infection and/or direct interaction with pathogens.


Assuntos
Âmnio/metabolismo , Corioamnionite/patologia , Ruptura Prematura de Membranas Fetais/metabolismo , Galectina 3/biossíntese , Âmnio/patologia , Proteínas Sanguíneas , Estudos de Casos e Controles , Corioamnionite/genética , Corioamnionite/metabolismo , Feminino , Ruptura Prematura de Membranas Fetais/genética , Galectinas/biossíntese , Humanos , Trabalho de Parto Prematuro/metabolismo , Gravidez , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA