Avaliação da expressão proteica em exossomos de sangue periférico de gestantes em Trabalho de Parto Pré-Termo e Rotura Prematura de Membranas Pré-Termo
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Data
2020-02-19
Autores
Tronco, Júlia Abbade
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A gestação é um período caracterizado por diversas alterações fisiológicas, hormonais e imunológicas do organismo materno. Em condições fisiológicas, entre a 37ª e 40ª semana de gestação, a sequência de eventos que levam ao parto inclui o aumento da contratilidade uterina, dilatação cervical e ruptura das membranas corioamnióticas. O trabalho de parto pré-termo com bolsa íntegra (TPP) é definido pelo trabalho de parto que ocorre antes da 37ª semana de gestação. O parto pré-termo é uma consequência desse desfecho e é a causa principal de morbidade e mortalidade neonatal. A Rotura Prematura de Membranas Pré-termo (RPM-PT) é outra complicação obstétrica, que ocorre em 3% de todos as gestações e pode ser identificada em 30-40% dos casos de TPP. Tanto o TPP quanto a RPM-PT possuem etiologia ainda pouco elucidada. Apesar da compreensão de algumas vias moleculares ativadoras destes desfechos adversos e dos diversos estudos sobre este tema, não existem biomarcadores capazes de predizer tais condições. Nesse contexto, os exossomos desempenham um papel como mediadores de comunicação intercelular e podem atuar na gestação e influenciar seu desfecho. Além disso, seu conteúdo, composto por proteínas, lipídios e RNAs tem sido bastante estudado como potenciais biomarcadores de diversas doenças. No campo da prematuridade, a relação entre os exossomos ou o conteúdo deles e o parto pré-termo ainda é pouco elucidada. Portanto, o objetivo do estudo foi avaliar a expressão proteica de exossomos isolados de sangue periférico de gestantes em Trabalho de Parto Pré-termo (TPP) e Rotura de Membranas Pré-termo (RPM-PT). Para tal, foram coletadas amostras de sangue periférico de pacientes no terceiro trimestre de gestação em TPP, RPM-PT, Termo em trabalho de parto (T+TP) e Termo fora de trabalho de parto (T). Os exossomos foram isolados a partir do plasma obtido, caracterizados por análise de partículas, quantificação proteica e Western Blot (WB) e, posteriormente, essas amostras foram submetidas à avaliação da expressão das proteínas de interesse (Hemopexina, Alfa-2-macroglobulina e C1INH) por WB. Por meio de análise semi-quantitativa foi possível observar um aumento da expressão da proteína Alfa-2-macroglobulina nos grupos Pré-termo (TPP + RPM-PT) em relação aos grupos Termo (T+TP e T) [1.07+- 0.30 vs 0.42 +- 0.17, p<0.0001]. A Alfa-2-macroglobulina é uma proteína envolvida em processos inflamatórios e no remodelamento de matriz extracelular. Considerando que o parto pré-termo é uma complicação com base fisiopatológica em processos inflamatórios e de remodelação dos tecidos gestacionais, o achado obtido no estudo é de importante relevância e pode contribuir para a literatura no contexto dos exossomos e prematuridade.
Pregnancy is a period characterized by many physiological, hormonal and immunological changes on maternal organism. On physiological conditions, between the 37th and 40th week of pregnancy, the sequence of events that lead to birth include the increase of uterine contractility, cervical dilatation and rupture of chorioamniotic membranes. Preterm labor (PTL) is defined by labor happening before the 37th week of pregnancy. Preterm birth is the consequence of this obstetrical complication and is the main cause of neonatal morbidity and mortality. Preterm premature rupture of membranes (PPROM) is another obstetrical complication, affects 3% of all pregnancies and can be identified in 30-40% of PTL cases. Both PTL and PPROM have multifactorial etiology and is still poorly elucidated. Despite the comprehension of many molecular pathways that can activate those adverse pregnancy outcomes and the numerous studies about this subject, there are no biomarkers capable of predicting such complications. In this context, the exosomes present on maternal circulation have the potential of representing the utero-placental environment from samples obtained by non-invasively ways. They are secreted by almost any cell types and are present on body fluids. Recent studies showed that this nanovesicles play an important role of intercellular communication mediators and can act on pregnancy and influence its outcome. Besides that, its content, composed by proteins, lipids and RNAs, have been studied as potential biomarkers of many diseases. On the prematurity context, the relation between exosomes, its content and preterm birth is still poorly understood. In this context, the aim of this study was to evaluate the exosome’s protein expression isolated from peripheral blood of women with preterm labor (PTL) and preterm premature rupture of membranes (PPROM). Therefore, samples of peripheral blood from patients with PTL, PPROM, term in labor (TL) and term out of labor (T) were collected. Exosomes were isolated from plasma and characterized by nanoparticle analysis, protein quantification and Western Blot (WB). Further, a WB was performed to identify the proteins of interest (hemopexin, alpha-2-macroglobulin and C1 inhibitor). By semi-quantitative analysis it was possible to observe a higher expression of alpha-2-macroglobulin on preterm groups (PTL + PPROM) when compared to term groups (TL + T) [1.07 +- 0.30 vs 0.42 +- 0.17, p<0.0001]. Alpha-2-macroglobulin is a protein involved in inflammatory processes and on extracellular matrix remodeling. Considering that preterm birth is a complication with a pathophysiological basis on inflammatory processes and gestational tissues remodeling, the obtained data of this study is relevant and can contribute to the knowledge on the context of exosomes and prematurity.
Pregnancy is a period characterized by many physiological, hormonal and immunological changes on maternal organism. On physiological conditions, between the 37th and 40th week of pregnancy, the sequence of events that lead to birth include the increase of uterine contractility, cervical dilatation and rupture of chorioamniotic membranes. Preterm labor (PTL) is defined by labor happening before the 37th week of pregnancy. Preterm birth is the consequence of this obstetrical complication and is the main cause of neonatal morbidity and mortality. Preterm premature rupture of membranes (PPROM) is another obstetrical complication, affects 3% of all pregnancies and can be identified in 30-40% of PTL cases. Both PTL and PPROM have multifactorial etiology and is still poorly elucidated. Despite the comprehension of many molecular pathways that can activate those adverse pregnancy outcomes and the numerous studies about this subject, there are no biomarkers capable of predicting such complications. In this context, the exosomes present on maternal circulation have the potential of representing the utero-placental environment from samples obtained by non-invasively ways. They are secreted by almost any cell types and are present on body fluids. Recent studies showed that this nanovesicles play an important role of intercellular communication mediators and can act on pregnancy and influence its outcome. Besides that, its content, composed by proteins, lipids and RNAs, have been studied as potential biomarkers of many diseases. On the prematurity context, the relation between exosomes, its content and preterm birth is still poorly understood. In this context, the aim of this study was to evaluate the exosome’s protein expression isolated from peripheral blood of women with preterm labor (PTL) and preterm premature rupture of membranes (PPROM). Therefore, samples of peripheral blood from patients with PTL, PPROM, term in labor (TL) and term out of labor (T) were collected. Exosomes were isolated from plasma and characterized by nanoparticle analysis, protein quantification and Western Blot (WB). Further, a WB was performed to identify the proteins of interest (hemopexin, alpha-2-macroglobulin and C1 inhibitor). By semi-quantitative analysis it was possible to observe a higher expression of alpha-2-macroglobulin on preterm groups (PTL + PPROM) when compared to term groups (TL + T) [1.07 +- 0.30 vs 0.42 +- 0.17, p<0.0001]. Alpha-2-macroglobulin is a protein involved in inflammatory processes and on extracellular matrix remodeling. Considering that preterm birth is a complication with a pathophysiological basis on inflammatory processes and gestational tissues remodeling, the obtained data of this study is relevant and can contribute to the knowledge on the context of exosomes and prematurity.
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Palavras-chave
Vesículas Extracelulares, Exossomos, Trabalho de Parto Pré-termo, Rotura Prematura de Membranas Pré-termo, Parto Pré-termo, Prematuridade, Gestação, Exosomes, Extracellular vesicles, Prematurity