An increase in elastogenic components in the placental villi of women with chronic venous disease during pregnancy is associated with decreased EGFL7 expression level

Abstract

Chronic venous disease (CVD) is the response to a series of hemodynamic changes in the venous system and the onset of this disease is often triggered by pregnancy. Placental tissue is particularly sensitive to the characteristic changes which occurs in venous hypertension. In this regard, changes in the extracellular matrix (ECM), that occur to adapt to this situation, are fundamental to controlling elastogenesis. Therefore, the aim of the present study was to analyze the changes that occur in the mRNA and protein expression level of proteins related to elastogenesis in the placental villi of women diagnosed with CVD, in the third trimester of pregnancy. An observational, analytical and prospective cohort study was conducted, in which the placenta from 62 women with CVD were compared with that in placenta from 52 women without a diagnosis of CVD. Gene and protein expression levels were analyzed using reverse transcription-quantitative PCR and immunohistochemistry, respectively. The results showed a significant decrease in the gene and protein expression level of EGFL7 in the placental villi of women with CVD. By contrast, significant increases in the gene and protein expression level of ECM-related proteins, such as tropoelastin, fibulin 4, fibrillin 1 and members of the lysyl oxidase family (LOX and LOXL-1) were also found in the placental villi of women with CVD. To the best of our knowledge, the results from the present study showed for the first time that CVD during pregnancy was associated with changes in the mRNA and protein expression level in essential components of the EGFL7-modulated elastogenesis process in placental villi.

Chronic Venous Disease in Pregnant Women Causes an Increase in ILK in the Placental Villi Associated with a Decrease in E-Cadherin

“…For the first time, we have demonstrated significant changes in tissue expression of critical cell to ECM (ILK) and cell to cell (e-cadherin, cadherin 7 and cadherin-9) components in placentas of women with CVD. These results are in consonance with our previous studies that showed that CVD is associated with a set of changes in the placenta in the ECM [7,16] and cell behavior [13,17].…”

“…In this line, we have demonstrated that maternal venous dysfunction also induces multiple abnormalities and damage in the placenta, evidencing an increase of hypoxic markers and enhanced apoptosis [13], markers of oxidative stress [14] and altered angiogenesis and lymphangiogenesis [15]. Moreover, we also found that CVD is associated with changes in the placental composition [7,16] and signaling [17], suggesting that this condition drives detrimental modifications in the placental structure and functioning, probably representing a unique pathophysiological feature in response to the venous dysfunction.…”

“…RNA samples at a concentration of 50 ng/µL were used to synthesize complementary DNA (cDNA) by reverse transcription (RT). 4 µL of each sample is mixed with 4 µL of oligo-dT 0.25 µg/µL solution (Thermo Fisher Scientific, Inc., Waltham, MA, USA), and incubated at 65 • C for 10 min in a dry bath (AccuBlock, Labnet International Inc., NJ, USA) to denature the RNA [16,29]. Hereunder, samples were put on ice and 10 µL of a reverse transcription mix containing the following products was added for each sample: 2.8 µL First Strand Buffer 5X (250 mM Tris-HCl and pH 8.3; 375 mM KCl; 15 mM MgCl2) (Thermo Fisher Scientific, Inc., Waltham, MA, USA); 2 µL of 10 mM deoxyribonucleotides triphosphate; 2 µL of 0.1 M dithiothreitol; 1.7 µL of DNase-and RNase-free water; 0.5 µL of RNase inhibitor (RNase Out); 1 µL of reverse transcriptase enzyme (all from Thermo Fisher Scientific, Inc., Waltham, MA, USA).…”

The Pivotal Role of the Placenta in Normal and Pathological Pregnancies: A Focus on Preeclampsia, Fetal Growth Restriction, and Maternal Chronic Venous Disease

“…All of these molecular variations may also be related to a switch in lipidomic profiling in the placenta of women with CVeD [269]. Finally, we observed that profound remodeling affects the placental tissue, with dysregulation of collagen and elastic fibers, including an increase in type III collagen, matrix metalloprotease 9 (MMP-9), and elastogenic processes with a decrease in EGFL7 expression [270,271]. Currently, the impact of CVeD in maternal and fetal well-being is starting to be elucidated.…”

Contribution of the Elastic Component and Venous Wall Arterialization in Patients with Venous Reflux

“…The last steps were dehydration in 96% alcohol for 5 min, dehydration in 100% alcohol for 5 min, xylol clearance for 10 min and Cytoseal™ mounting (Epredia™ 8310-4, Thermo Fisher Scientific A., Waltham, MA, USA). The use of this technique evidences the presence of the elastic fibers in a brown color [16].…”