RESUMEN
Preeclampsia (PE) is a unique pregnancy disorder affecting women across the world. It is characterized by the new onset of hypertension with coexisting end-organ damage. Although the disease has been known for centuries, its exact pathophysiology and, most importantly, its prevention remain elusive. The basis of its associated molecular changes has been attributed to the placenta and the hormones regulating its function. One such hormone is chromogranin A (CgA). In the placenta, CgA is cleaved to form a variety of biologically active peptides, including catestatin (CST), known inter alia for its vasodilatory effects. Recent studies indicate that the CST protein level is diminished both in patients with hypertension and those with PE. Therefore, the aim of the present paper is to review the most recent and most relevant in vitro, in vivo, and clinical studies to provide an overview of the proposed impact of CST on the molecular processes of PE and to consider the possibilities for future experiments in this area.
Asunto(s)
Hipertensión , Preeclampsia , Humanos , Femenino , Cromogranina A/metabolismo , Fragmentos de Péptidos/metabolismoRESUMEN
Pregnancy is a state of physiological and hormonal changes. One of the endocrine factors involved in these processes is chromogranin A, an acidic protein produced, among others, by the placenta. Although it has been previously linked to pregnancy, no existing articles have ever managed to clarify the role of this protein regarding this subject. Therefore, the aim of the present study is to gather knowledge of chromogranin A's function with reference to gestation and parturition, clarify elusive information, and, most importantly, to formulate hypotheses for the future studies to verify.
Asunto(s)
Cromograninas , Sistema Endocrino , Embarazo , Femenino , Humanos , Cromogranina A/metabolismo , Cromograninas/metabolismo , Sistema Endocrino/metabolismo , Parto , Placenta/metabolismo , Fragmentos de Péptidos/metabolismoRESUMEN
One of the most dangerous complications of pregnancy is preeclampsia (PE), a disease associated with a high risk of maternal and fetal mortality and morbidity. Although its etiology remains unknown, the placenta is believed to be at the center of ongoing changes. One of the hormones produced by the placenta is chromogranin A (CgA). Thus far, its role in pregnancy and pregnancy-related disorders is enigmatic, yet it is known that both CgA and its derived peptide catestatin (CST) are involved in the majority of the processes that are disturbed in PE, such as blood pressure regulation or apoptosis. Therefore, in this study, the influence of the preeclamptic environment on the production of CgA using two cell lines, HTR-8/SVneo and BeWo, was investigated. Furthermore, the capacity of trophoblastic cells to secrete CST to the environment was tested, as well as the correlation between CST and apoptosis. This study provided the first evidence that CgA and CST proteins are produced by trophoblastic cell lines and that the PE environment has an impact on CST protein production. Furthermore, a strong negative correlation between CST protein level and apoptosis induction was found. Hence, both CgA and its derived peptide CST may play roles in the complex process of PE pathogenesis.
Asunto(s)
Preeclampsia , Embarazo , Femenino , Humanos , Preeclampsia/metabolismo , Trofoblastos/metabolismo , Placenta/metabolismo , Línea Celular , Presión SanguíneaRESUMEN
Preeclampsia (PE) is a pregnancy-specific disorder affecting 4-10% of all expectant women. It greatly increases the risk of maternal and foetal death. Although the main symptoms generally appear after week 20 of gestation, scientific studies indicate that the mechanism underpinning PE is initiated at the beginning of gestation. It is known that the pathomechanism of preeclampsia is strongly related to inflammation and oxidative stress, which influence placentation and provoke endothelial dysfunction in the mother. However, as of yet, no "key players" regulating all these processes have been discovered. This might be why current therapeutic strategies intended for prevention or treatment are not fully effective, and the only effective method to stop the disease is the premature induction of delivery, mostly by caesarean section. Therefore, there is a need for further research into new pharmacological strategies for the treatment and prevention of preeclampsia. This review presents new preventive methods and therapies for PE not yet recommended by obstetrical and gynaecological societies. As many of these therapies are in preclinical studies or under evaluation in clinical trials, this paper reports the molecular targets of the tested agents or methods.
Asunto(s)
Preeclampsia , Embarazo , Femenino , Humanos , Preeclampsia/prevención & control , Preeclampsia/diagnóstico , Cesárea , Placentación , Estrés Oxidativo , Receptor 1 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
In vivo studies on the pathology of gestation, including preeclampsia, often use small mammals such as rabbits or rodents, i.e., mice, rats, hamsters, and guinea pigs. The key advantage of these animals is their short reproductive cycle; in addition, similar to humans, they also develop a haemochorial placenta and present a similar transformation of maternal spiral arteries. Interestingly, pregnant dams also demonstrate a similar reaction to inflammatory factors and placentally derived antiangiogenic factors, i.e., soluble fms-like tyrosine kinase 1 (sFlt-1) or soluble endoglin-1 (sEng), as preeclamptic women: all animals present an increase in blood pressure and usually proteinuria. These constitute the classical duet that allows for the recognition of preeclampsia. However, the time of initiation of maternal vessel remodelling and the depth of trophoblast invasion differs between rabbits, rodents, and humans. Unfortunately, at present, no known animal replicates a human pregnancy exactly, and hence, the use of rabbit and rodent models is restricted to the investigation of individual aspects of human gestation only. This article compares the process of placentation in rodents, rabbits, and humans, which should be considered when planning experiments on preeclampsia; these aspects might determine the success, or failure, of the study. The report also reviews the rodent and rabbit models used to investigate certain aspects of the pathomechanism of human preeclampsia, especially those related to incorrect trophoblast invasion, placental hypoxia, inflammation, or maternal endothelial dysfunction.
Asunto(s)
Preeclampsia , Conejos , Femenino , Embarazo , Humanos , Ratones , Ratas , Cobayas , Animales , Receptor 1 de Factores de Crecimiento Endotelial Vascular , Placenta/irrigación sanguínea , Roedores , Reproducibilidad de los ResultadosRESUMEN
BACKGROUND: Although preeclampsia has long been recognized as a condition affecting late pregnancy, little is known of its pathogenesis or treatment. The placenta releases a number of hormones and molecules that influence the course of pregnancy, one of which is chromogranin A, a soluble protein secreted mainly from the chromaffin cells of the adrenal medulla. Its role in pregnancy and pregnancy-related disorders remains unclear. Therefore, the main aim of the proposed study is to determine whether chromogranin A is related with the occurrence of preeclampsia. METHODS: Placental samples were collected from 102 preeclamptic patients and 103 healthy controls, and Chromogranin A gene (CHGA) expression was measured using real-time RT-PCR, The RT-PCR results were verified on the protein level using ELISA. The normal distribution of the data was tested using the Shapiro-Wilk test. The clinical and personal characteristics of the groups were compared using the Student's t-test for normally-distributed data, and the χ2 test for categorical variables. The Mann-Whitney U test was used for non-normally distributed data. As the log- transformation was not suitable for the given outcomes, the Box- Cox Transformation was used to normalize data from ELISA tests and CHGA expression. Values of P < .05 were considered statistically significant. RESULTS: Chromogranin A gene expression was found to be significantly higher in the study group than in controls. Protein analyses showed that although the CgA concentration in placental samples did not differ significantly, the catestatin (CST) level was significantly lower in samples obtained from women with preeclampsia, according to the controls. CONCLUSIONS FOR PRACTICE: This study for the first time reveals that chromogranin A gene expression level is associated with preeclampsia. Moreover, the depletion in catestatin level, which plays a protective role in hypertension development, might be a marker of developing preeclampsia. Further studies may unravel role of Chromogranin A in the discussed disease.
Asunto(s)
Cromogranina A/metabolismo , Fragmentos de Péptidos/metabolismo , Placenta/metabolismo , Preeclampsia/metabolismo , Biomarcadores/metabolismo , Estudios de Casos y Controles , Cromogranina A/genética , Femenino , Expresión Génica , Humanos , Fragmentos de Péptidos/genética , Preeclampsia/genética , EmbarazoRESUMEN
Preeclampsia is a pregnancy disorder associated with shallow placentation, forcing placental cells to live in hypoxic conditions. This activates the transcription factor kappa B (NFκB) in maternal and placental cells. Although the role of NFκB in preeclampsia is well documented, its mechanism of activation in trophoblastic cells has been never studied. This study investigates the mechanism of NFκB activation in a first trimester trophoblastic cell line (HTR8/SVneo) stimulated by a medium containing serum from preeclamptic (PE) or normotensive (C) women in hypoxic (2% O2) or normoxic (8% O2) conditions. The results indicate that in HTR8/SVneo cells, the most widely studied NFκB pathways, i.e., canonical, non-canonical and atypical, are downregulated in environment PE 2% O2 in comparison to C 8% O2. Therefore, other pathways may be responsible for NFκB activation. One such pathway depends on the activation of NFκB by the p53/RSK1 complex through its phosphorylation at Serine 536 (pNFκB Ser536). The data generated by our study show that inhibition of the p53/RSK1 pathway by p53-targeted siRNA results in a depletion of pNFκB Ser536 in the nucleus, but only in cells incubated with PE serum at 2% O2. Thus, the p53/RSK1 complex might play a critical role in the activation of NFκB in trophoblastic cells and preeclamptic placentas.
Asunto(s)
FN-kappa B/metabolismo , Preeclampsia/patología , Proteínas Quinasas S6 Ribosómicas 90-kDa/metabolismo , Trofoblastos/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Hipoxia de la Célula/fisiología , Línea Celular , Activación Enzimática/genética , Femenino , Humanos , Placenta/patología , Embarazo , Interferencia de ARN , ARN Interferente Pequeño/genética , Especies Reactivas de Oxígeno/metabolismo , Proteína p53 Supresora de Tumor/genéticaRESUMEN
Although higher nuclear factor κB (NFκB) expression and activity is observed in preeclamptic placentas, its mechanism of activation is unknown. This is the first study to investigate whether the canonical, non-canonical, or atypical NFκB activation pathways may be responsible for the higher activation of NFκB observed in preeclamptic placentas. The study included 268 cases (130 preeclamptic women and 138 controls). We studied the expression of the genes coding for NFκB activators (NIK, IKKα, IKKß, and CK2α) and inhibitors (IκBα and IκBß) using RT-PCR in real time. The RT-PCR results were verified on the protein level using ELISA and Western blot. To determine the efficiency of the pathways, the ratios of activator(s) to one of the inhibitors (IκBα or IκBß) were calculated for each studied pathway. The preeclamptic placentas demonstrated significantly lower IKKα and CK2α but higher IκBα and IκBß protein levels. In addition, the calculated activator(s) to inhibitor (IκBα or IκBß) ratios suggested that all studied pathways might be downregulated in preeclamptic placentas. Our results indicate that preeclamptic placentas may demonstrate mechanisms of NFκB activation other than the canonical, non-canonical, and atypical forms. In these mechanisms, inhibitors of NFκB may play a key role. These observations broaden the existing knowledge regarding the molecular background of preeclampsia development.
Asunto(s)
Quinasa I-kappa B/genética , Preeclampsia/genética , Proteínas Serina-Treonina Quinasas/genética , Adulto , Núcleo Celular/genética , Femenino , Regulación de la Expresión Génica/genética , Humanos , FN-kappa B/genética , Placenta/metabolismo , Placenta/patología , Preeclampsia/patología , Embarazo , Transducción de Señal/genética , Quinasa de Factor Nuclear kappa BRESUMEN
A linkage of dichorionic (DC) twin pregnancies with selective intrauterine growth restriction (IUGR) to alterations in placental gene expression is unclear. The aim of the study was to identify placental genes related to hypoxia, adipogenesis and human growth which may contribute to IUGR development. The study group (IUGR/AGA) comprised dichorionic (DC) twin pregnancies, where the weight of the twins differed by > 15%; in addition, one twin was small for gestational age (< 10th percentile-SGA) (IUGR) while the other was appropriate for gestational age (> 10th percentile-AGA). In the control group (AGA/AGA), both fetuses were AGA and their weights differed by < 15%. In the first step (selection), placental expression of 260 genes was analysed by commercial PCR profiler array or qPCR primer assay between six pairs of IUGR/AGA twins. In the second stage (verification), the expression of 20 genes with fold change (FC) > 1.5 selected from the first stage was investigated for 75 DC pregnancies: 23 IUGR/AGA vs. 52 AGA/AGA. The expression of Angiopoetin 2, Leptin and Kruppel-like factor 4 was significantly higher, and Glis Family Zinc Finger 3 was lower, in placentas of SGA fetuses (FC = 3.3; 4.4; 1.6; and - 1.8, respectively; p < 0.05). The dysregulation of gene expression related to angiogenesis and growth factors in placentas of twins born from IUGR/AGA pregnancies suggest that these alternations might represent biological fetal adaptation to the uteral condition. Moreover, DC twin pregnancies may be a good model to identify the differences in placental gene expression between SGA and AGA fetuses.
Asunto(s)
Retardo del Crecimiento Fetal/genética , Perfilación de la Expresión Génica/métodos , Placenta/metabolismo , Embarazo Gemelar/genética , Proteínas de Unión al ADN , Femenino , Edad Gestacional , Humanos , Hipoxia , Recién Nacido , Recién Nacido Pequeño para la Edad Gestacional , Factor 4 Similar a Kruppel , Factores de Transcripción de Tipo Kruppel/genética , Masculino , Embarazo , Proteínas Represoras , Transactivadores , Factores de Transcripción/genéticaRESUMEN
PURPOSE OF REVIEW: Preeclampsia (PE) is a hypertensive disorder exclusive for pregnancy. It affects women all over the world and poses a great threat to life, both for mother and child. No definitive treatment exists and placenta delivery comprises the only known cure for PE. One of the most severe complications observed in preeclamptic women is the occurrence of cardiovascular diseases (CVDs) later in life. RECENT FINDINGS: Both PE and CVDs share some of their pathogenic pathways and gene variations. Thus far, a number of publications have examined those relationships; however, almost all of them focus only on common risk factors. The precise pathomechanism and genetic basis of PE and its associated cardiovascular complications remain unknown. Therefore, the aim of this review is to unify and clarify the current state of knowledge and provide direction for future studies, especially those regarding the genetic aspect.
Asunto(s)
Enfermedades Cardiovasculares , Enfermedades Cardiovasculares/genética , Enfermedades Cardiovasculares/metabolismo , Enfermedades Cardiovasculares/fisiopatología , Femenino , Antecedentes Genéticos , Humanos , Preeclampsia/genética , Preeclampsia/metabolismo , Preeclampsia/fisiopatología , Embarazo , Factores de RiesgoRESUMEN
BACKGROUND: The mechanism of preeclampsia and its way of inheritance are still a mystery. Biochemical and immunochemical studies reveal a substantial increase in tumor necrosis factor alpha, interleukin-1 beta, and interleukin-6 concentrations in the blood of women with preeclampsia. The level of these factors is regulated by nuclear facxtor-kappa B, whose activation in a classical pathway requires inhibitory kappa B kinase gamma (known as NEMO or IKBKG). Moreover, NEMO can schedule between cytoplasma and the nucleus. In the nucleus, IKBKG interacts with other proteins, and thus, it is implicated in the regulation of different gene expressions, which are related to cell cycle progression, proliferation, differentiation, and apoptosis. OBJECTIVE: This is the first study investigating the association between the level of NEMO gene expression and the presence of preeclampsia. We tested the hypothesis that the simultaneous increase in NEMO gene expression both in the mother and her fetus may be responsible for the preeclampsia development. Moreover, the relationships between clinical risk factors of preeclampsia and the levels of NEMO gene expression in blood, umbilical cord blood, and placentas were investigated. STUDY DESIGN: A total of 91 women (43 preeclamptic women and 48 controls) and their children were examined. Real-time reverse transcription-polymerase chain reaction was used to assess the amount total NEMO messenger ribonucleic acid (mRNA) content and the mRNA level of each NEMO transcript from exons 1A, 1B, and 1C in maternal blood, umbilical cord blood, and placentas. Univariate analyses and correlation tests were performed to examine the association between NEMO gene expression and preeclampsia. RESULTS: Newborn weight and height, maternal platelet number, and gestational age (week of delivery) were lower in the group of women with preeclampsia than controls. NEMO gene expression level was found to be almost 7 times higher in the group of women with preeclampsia than healthy controls. The correlation analysis found that a simultaneous increase in the expression level of total NEMO mRNA in maternal blood and the mRNA for total NEMO (Rs = 0.311, P < .05), transcripts 1A (Rs = 0.463, P < .01), 1B (Rs = 0.454, P < .01), and 1C (Rs = 0.563, P < .001) in fetal blood was observed in preeclamptic pregnancies. In addition, the mRNA levels for total NEMO and transcripts 1A, 1B, and 1C were lower in placentas derived from pregnancies complicated by preeclampsia. CONCLUSION: Simultaneous increase of NEMO gene expression in maternal and fetal blood seems to be relevant for preeclampsia development. The results of our study also suggest that a decreased NEMO gene expression level in preeclamptic placentas may be the main reason for their intensified apoptosis.
Asunto(s)
Quinasa I-kappa B/genética , Quinasa I-kappa B/metabolismo , Preeclampsia/sangre , Preeclampsia/genética , ARN Mensajero/metabolismo , Adulto , Biomarcadores/metabolismo , Estudios de Casos y Controles , Femenino , Sangre Fetal/metabolismo , Expresión Génica , Humanos , Placenta/metabolismo , Reacción en Cadena de la Polimerasa , Embarazo , Transcripción GenéticaRESUMEN
BACKGROUND: Essential hypertension (EH) is the most common cardiovascular disease worldwide, and it has a strong genetic component. Cortisol homeostasis is an important factor in controlling blood pressure, and the availability of this hormone is regulated by 11ßhydroxysteroid dehydrogenase type 1 enzyme (11ßHSD1), which converts cortisone into cortisol. MATERIALS AND METHODS: We investigated the correlation between EH and the single nucleotide polymorphism (SNP) ins4436A located on the hydroxysteroid (11-beta) dehydrogenase 1 gene among the Polish population. The study included a total of 268 patients with confirmed EH and 151 unrelated controls. All studied polymorphisms were detected using the restriction fragment length polymorphism (RFLP) method. RESULTS: The carriage of ins4436A (rs45487298) polymorphism in intron 3 of the HSD11B1 gene was more frequent among patients with EH than among controls (p=0.013). The analysis of association of ins4436A with the risk of EH indicated an odds ratio (OR) of 2.44 (95% confidential interval: 1.24-4.82). Moreover, essential hypertension occurred less frequently in males than in females. Results of multivariate analysis in the study group showed that ins4436A is a strong predictor of diabetes mellitus type 2 and ins4436A may lead to a decrease of the high-density lipoprotein (HDL) cholesterol level. DISCUSSION: The cause of essential hypertension has not been fully established, but genetic factors seem to play a very important role. In our study we found that ins4436A in the HSD11B1 gene was associated with essential hypertension in a Polish population. Nevertheless, the impact of ins4436A in the HSD11B1 gene on the occurrence of essential hypertension requires further investigations.
Asunto(s)
11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1/genética , Predisposición Genética a la Enfermedad , Hipertensión/genética , Polimorfismo de Nucleótido Simple , Población Blanca/genética , Anciano , HDL-Colesterol/sangre , Diabetes Mellitus Tipo 2/enzimología , Diabetes Mellitus Tipo 2/genética , Hipertensión Esencial , Femenino , Humanos , Hipertensión/enzimología , Masculino , Persona de Mediana Edad , Polonia , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
This study investigates the potential role of 17 chosen polymorphisms in 15 candidate genes and the risk of myocardial infarction in patients under 45 years of age. The study consists of 271 patients with myocardial infarction and 141 controls. The analysis of genetic polymorphisms was performed using the PCR-RFLP method. Of the chosen polymorphisms, two (Leu125Val PECAM1 and A1/A2 FVII) are related to myocardial infarction and two (C677T MTHFR and 5A/6A MMP3) to advanced stenosis in arterial vessels (> 75%). We also found that the frequency of some combinations among the analyzed genes and environmental factors varied between the patient and control groups.
Asunto(s)
Infarto del Miocardio/genética , Polimorfismo Genético , Adulto , Factores de Edad , Aterosclerosis/etiología , Aterosclerosis/genética , Femenino , Tamización de Portadores Genéticos , Humanos , Hipertensión/genética , Masculino , Metaloproteinasa 3 de la Matriz/genética , Metilenotetrahidrofolato Reductasa (NADPH2)/genética , Persona de Mediana Edad , Infarto del Miocardio/etiología , Factores de Riesgo , Fumar/genéticaRESUMEN
BACKGROUND: A quantitative history using Calgary syncope syndrome score (CSSS) is able to define the likely cause of syncope, but there is still a lack of diagnostic screening tests for vasovagal syncope (VVS). The aim of the present study was to develop a screening test for VVS on the basis of CSSS and the relationship between polymorphic variants of the G-system signaling protein genes and tilting results. METHODS AND RESULTS: From 730 syncopal patients, 307 consecutive subjects without structural and electrical abnormalities were genotyped and examined on blood pressure (BP) and tilt testing. Genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism in genes encoding Gsα-protein GNAS1 (rs7121), G-protein ß 3 subunit (rs5443) and the cardiac regulator of G-protein signaling RGS2 (rs4606). The control group consisted of 100 healthy volunteers with a negative history of syncope. From multivariate regression analysis, being a carrier of 393T GNAS1 (odds ratio [OR], 2.29) and systolic BP (OR, 0.98) remained as independent factors associated with positive tilt results. The resultant screening test for VVS consisted of the following: carrier of 393T GNAS1; systolic BP < 131 mm Hg (from the receiver operating characteristic [ROC] curve); and CSSS ≥-2. Using ROC curve analysis for systolic BP and CSSS, 2 final models for the screening test were constructed: highest sensitivity (89%) and highest specificity (99%). CONCLUSIONS: The novel screening test including the variation of Gsα protein gene seems to be helpful to identify tilt-induced vasovagal patients.
Asunto(s)
Subunidades alfa de la Proteína de Unión al GTP Gs/genética , Polimorfismo de Longitud del Fragmento de Restricción , Proteínas RGS/genética , Síncope Vasovagal/genética , Adulto , Presión Sanguínea/genética , Cromograninas , Humanos , Técnicas In Vitro , Masculino , Persona de Mediana Edad , Curva ROC , Análisis de Regresión , Síncope Vasovagal/fisiopatologíaRESUMEN
Stable gene integration and rapid selection of high-expressing clones are important when developing biopharmaceutical systems to produce a protein of interest. According to regulatory guidelines, the final production clones should be stable through multiple cell generations. To achieve long-term stable expression of Fab genes via recombinase-mediated cassette exchange (RMCE), we modified mutual configurations of the lox sequences. By inversion of the spacer orientation, we avoided the loss of the integrated gene after several dozen cycles of cell division. This feature also prevents reversible transgene integration. Although the RMCE allows us to generate transgenic lines rapidly relative to current methods, it remains difficult to obtain stable industrial cell lines for long-term culturing and for the initial development stage. In this study, we present an approach to shortening the timeline for therapeutic protein development. Our approach provides easy access to the same clonal cell line in the initial development phase, and also for the production of biopharmaceutical proteins.
Asunto(s)
Productos Biológicos , Línea Celular , Integrasas/genética , TransgenesRESUMEN
The influence of the glycosylation profile of IgG on biological activity is known, but it is not clear which glycoforms have the highest impact on the main mechanism of action. The aim of this study was to design a mathematical model for predicting the antibody-dependent cellular cytotoxicity (ADCC) activity and the Fc gamma IIIa receptors' (FcɣRIIIa) relative binding of rituximab drug products based on their glycosylation profile. An additional goal was to identify the glycoforms that have the greatest impact on these mechanisms of action. For these purposes, the glycosylation profile was examined by hydrophilic interaction ultra-performance liquid chromatography (HILIC-UPLC), ADCC was assessed using a Promega kit, and FcɣRIIIa's binding affinity was assessed by surface plasmon resonance (SPR) analysis of a group of >50 rituximab drug products. Based on the results, mathematical models for the ADCC and FcɣRIIIa binding affinity prediction were designed using JMP 13.2.0. The quality of the model and the influence of sample size and heterogeneity on the reliability were verified. The results allow for the evaluation of rituximab drug products' activity based on their glycosylation profile and show that with a sufficiently large and differentiated dataset, it is possible to generate models for different monoclonal antibodies.
Asunto(s)
Anticuerpos Monoclonales , Citotoxicidad Celular Dependiente de Anticuerpos , Glicosilación , Reproducibilidad de los Resultados , Rituximab/metabolismoRESUMEN
AIMS: G-proteins signal transduction pathways play a basic role in cardiovascular reflexes. We hypothesized that the predisposition to reflex-mediated syncope may be associated with genetic variations in G-protein genes. The aim of this study was to evaluate the effect of three single-nucleotide polymorphisms in G-protein genes on tilting outcome in syncopal patients. METHODS AND RESULTS: A total of 217 syncopal patients free from any other disease were genotyped and examined related to tilting results. Genotyping was performed by polymerase chain reaction followed by restriction fragment length polymorphism in gene encoding the Gs-protein alpha-subunit (polymorphism C393T), the G-protein beta 3 subunit--GNB3 (polymorphism C825T)--and for the cardiac regulator of G-protein signalling RGS2 (polymorphism C1114G). In multivariate logistic regression analysis, the homozygotes 825TT GNB3 (OR 0.37; 95% CI 0.14-0.97; P < 0.05) and body mass index (OR 0.87; 95% CI 0.78-0.97; P = 0.005) were independently associated with a lower chance of positive tilting results. No relationship was found between Vasovagal Syncope International Study type of syncope and the studied genotypes or the carriage of the polymorphic alleles. CONCLUSIONS: An association between tilting results and C825T GNB3 polymorphism in syncopal patients was found. The syncopal homozygotes 825TT GNB3 had a significantly lower chance of syncope during tilt testing.
Asunto(s)
Proteínas de Unión al GTP/genética , Polimorfismo de Nucleótido Simple/genética , Síncope/diagnóstico , Síncope/genética , Pruebas de Mesa Inclinada/estadística & datos numéricos , Adolescente , Adulto , Anciano , Femenino , Predisposición Genética a la Enfermedad/epidemiología , Predisposición Genética a la Enfermedad/genética , Estudio de Asociación del Genoma Completo , Humanos , Masculino , Persona de Mediana Edad , Polonia/epidemiología , Prevalencia , Reproducibilidad de los Resultados , Medición de Riesgo/métodos , Factores de Riesgo , Sensibilidad y Especificidad , Síncope/epidemiología , Adulto JovenRESUMEN
BACKGROUND: Preeclamptic pregnancies often present an intensified inflammatory state associated with the nuclear activity of NFκB. NEMO is an essential regulator of nuclear factor kappa B (NFκB) in cytoplasmic and nuclear cellular compartments. The aim of the present study is to examine the level and localization of the NEMO protein in preeclamptic and nonpreeclamptic placentas. METHODS: The study includes 97 preeclamptic cases and 88 controls. NEMO distribution was analyzed immunohistochemically. Its localization in the nuclear and cytoplasmic fractions, as well as in total homogenates of placental samples, was studied by western blot and ELISA. RESULTS: The western blot and ELISA results indicate a significant difference in NEMO concentration in the total and nuclear fractions between preeclamptic and control samples (p < 0.01 and p < 0.001, respectively). In the cytoplasmic complement, similar levels of NEMO were found in preeclamptic and control placentas. In addition, immunohistochemical staining revealed that the NEMO protein is mainly localized in the syncytiotrophoblast layer, with controls demonstrating a stronger reaction with NEMO antibodies. This study also shows that the placental level of NEMO depends on the sex of the fetus. CONCLUSIONS: The depletion of the NEMO protein in the cellular compartments of placental samples may activate one of the molecular pathways influencing the development of preeclampsia, especially in pregnancies with a female fetus. A reduction of the NEMO protein in the nuclear fraction of preeclamptic placentas may intensify the inflammatory state characteristic for preeclampsia and increase the level of apoptosis and necrosis within preeclamptic placentas.
Asunto(s)
Quinasa I-kappa B/genética , Placenta/metabolismo , Preeclampsia/genética , Adulto , Biomarcadores/metabolismo , Estudios de Casos y Controles , Femenino , Humanos , Quinasa I-kappa B/metabolismo , Recién Nacido , Masculino , Preeclampsia/metabolismo , Embarazo , Factores SexualesRESUMEN
BACKGROUND: Metalloproteinases (MMPs) play a pivotal role during the process of trophoblast invasion and placentation. The appearance of five functional single-nucleotide polymorphisms (SNP) in the genes of the metalloproteinases most commonly implicated in the implantation process may influence the development of preeclampsia. METHODS: Blood samples were collected from 86 mothers and 86 children after preeclampsia and 85 mothers and 85 children with uncomplicated pregnancies. The distribution of genotypes for -1607 1G/2G MMP1, -735 C/T MMP2, -1306 C/T MMP2, -1171 5A/6A MMP3, and -1562C/T MMP9 polymorphisms was determined by RFLP-PCR. RESULTS: The occurrence of 1G/1G MMP1 or 5A/5A MMP3 genotype in the mother or 1G/1G MMP1 or 5A/6A MMP3 genotype in the child is associated with preeclampsia development. Moreover, simultaneous maternal and fetal 1G/1G homozygosity increases the risk of preeclampsia development 2.39-fold and the set of maternal 5A/5A and fetal 5A/6A MMP3 genotypes by over 4.5 times. No association between the carriage of studied MMP2 or MMP9 polymorphisms and the predisposition to preeclampsia was found. CONCLUSION: The maternal 1G/1G MMP1 and 5A/5A MMP3 and fetal 1G/1G MMP1 and 5A/6A MMP3 gene polymorphisms may be strong genetic markers of preeclampsia, occurring either individually or together.
Asunto(s)
Metaloproteinasas de la Matriz/genética , Polimorfismo de Nucleótido Simple , Preeclampsia/genética , Adulto , Estudios de Casos y Controles , Femenino , Humanos , Recién Nacido , Masculino , EmbarazoRESUMEN
OBJECTIVE: The G protein is responsible for signal intracellular transduction and participates in cardiovascular reflexes. C825T polymorphism of the gene encodes the B3 subunit of G protein (GNB3) and causes the increased intracellular signal transduction. The aim was the evaluation GNB3 C825T polymorphism manifestation in vasovagal patients with no other diseases. METHODS: In 68 positive tilted patients genomic DNA was extracted from blood using an extraction kit. The GNB3 C825T polymorphism was diagnosed by restriction of the PCR amplicon with BseDI (MBI Fermentas). All patients were genotyped and next analyzed in regard to typical vasovagal history. RESULTS: The prevalence of genotype CC was 38%. Genotypes CT and TT were found equally in 31% of cases. The C allele in comparison to the T allele appeared in 54% vs 46% (p>0.05). Typical vasovagal history was present in 83% of patients. The frequency of GNB3 825T allele was significantly higher in patients with non-typical vasovagal history than in group with typical history (p<0.001). CONCLUSIONS: Genotype CC GNB3 is the most popular in vasovagal patients. The predisposition to vasovagal syncope seems to be not associated with the GNB3 825T allele. Further studies are planned to clarify the genotype/phenotype relationship in vasovagal patients.