Browsing by Author "Almeida, Henrique"
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- Addressing the role of maternal age on the metabolic profile of the placentaPublication . Pinheiro, Ana R.; Rodrigues, Adriana; Matos, Liliana; Guedes-Martins, Luís; Almeida, Henrique; Silva, ElisabeteAfter the age of 35, during pregnancy, there is an increased risk of impaired placentation. Changes in uterine redox balance seem to play a role in deficient placentation. We hypothesized that this local redox dysregulation has a negative impact on the placenta metabolic profile. Thus, we aimed to study the placenta metabolic profile during reproductive aging and the effect of antioxidant supplementation. Placentas were collected from pregnant women aged between 22 and 41 years, and from mice of different ages (3 or 9 months). Additionally, 9-month-old mice were treated with apocynin (5 mM) in the drinking water (3 weeks prior to and during pregnancy). Semi-quantitative real-time PCR was carried out to assess the expression of glucose and fatty acid transporters. Quantitative results are presented with standard error of the mean (SEM). In the human study, Spearman’s correlation was used for the comparative analysis of the variables studied. In the animal model, Student’s t-test was used for comparative analysis of the variables studied. A p-value of less than 0.05 was considered statistically significant. Concerning glucose transport, the results show that the expression of glucose transporter 1 is strongly negatively correlated with maternal age (r = 0.710; p = 0.0121). Regarding lipid metabolism, there is a strong negative correlation between the fatty acid transporter 4 expression and maternal age (r = -0.6865, p = 0.0233). In mice, the expression of glucose transporter 1 was also decreased in 9-month-old females (p = 0.0329, when compared to the 3-month-old females group). No differences were observed between the reproductively aged females treated with apocynin and the young females (p = 0.1990). The results of this study show that reproductive aging is linked with changes in the placenta metabolic profile. In the mice study model, antioxidant supplementation attenuated the changes observed in nutrient transport in the placenta.
- Proteomics characterization of human uterine samples during reproductive ageingPublication . Dias, Bruna; Formoso, Inês; Guedes-Martins, Luís; Almeida, Henrique; Silva, Elisabete; Silva, André M.N.; Matos, LilianaNowadays, postponing motherhood has become more common in modern societies. Such decision rises major social and health concerns, as advanced maternal age associates with low fertility, pregnancy complications and a greater need for assisted reproductive techniques. Female reproductive ageing is associated with alterations that are particularly evident in the ovaries. Yet, given the pivotal contribution of the uterus to embryo adhesion, implantation, and fetal development, we believe that age-dependent uterine changes impair fertility. Evidence in mice already reported age-associated uterine changes. Also, albumin carbonylation was found increased in uterine samples from older term-pregnant women. Moreover, preliminary data show changes in composition and oxidation status of uterine extracellular matrix proteins with higher maternal age. We hypothesize that the uterine proteome changes during reproductive ageing, impairing tissue structure and function. This study aims to identify age-related alterations in uterine protein composition through a complete proteomics approach. Uterine samples were collected during c-section from termpregnant women and immediately fixed for paraffin-embedding or frozen for molecular analysis. Six samples were selected: three from younger and three from reproductively aged women. Paraffin-embedded samples were histologically analyzed after H+E staining. Protein lysates were obtained after sample homogenization and total protein content was determined and were then separated by SDS-PAGE, stained with Coomassie Blue, and will be subjected to a MS based proteomics study. Histological analysis of the samples has proven their uterine origin. SDS-PAGE results confirmed the successful protein extraction. The identification of uterine proteins differentially expressed according to maternal age will allow the development of therapeutic approaches, aiming to attenuate age-associated female fertility decline.
- Unveiling uterine senescence and its potential role in age-related female fertility declinePublication . Formoso, Inês; Guedes-Martins, Luís; Almeida, Henrique; Silva, Elisabete; Matos, LilianaNowadays, infertility is a major topic present in public health discussions due to its increased prevalence worldwide. With time, female infertility rates have increased, mainly, because women are delaying childbearing to their late 30s/early 40s, leading to fertility decline, complications during gestation, and a higher demand for assisted reproductive techniques. Uterine alterations have been pointed out as an important contributor, as uterine function is linked with the ability to conceive and carry a healthy pregnancy. Previous data reported uterine alterations in reproductively aged mice. Also, using human uterine samples, it was shown an age-related increase in albumin carbonylation and variations in the oxidative status of extracellular matrix proteins. We believe that cellular senescence contributes to uterine alterations, impacting tissue microenvironment and impairing its function. In this project, we aim to evaluate the appearance of senescence-related uterine alterations with increasing age. Uterine samples from term-pregnant women with ages between 20 and 41 were homogenized, protein lysates were submitted to western blot analysis for relative quantification of senescence-associated proteins, including markers of nuclear damage and Senescenceassociated secretory phenotype (SASP) constituents. Currently, our results show a strong and significant positive correlation between nuclear proteins, such as HP1, Lamin B1, and PH2AX, and age. Regarding pro-inflammatory proteins, MCP1 tended to increase with age while IL6 and IL1 did not change. Also, SASP proteins like MMP3 and PAI1 presented a weak negative correlation with age. In the uterus, reproductive ageing courses with increased senescence-associated nuclear alterations reflecting the existence of DNA damage and organelle destabilization. Further analysis is needed to confirm the absence of significant variations in SASP proteins.