Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCA337

Poster Communications


D. Pérez-Cremades1,2, S. Novella1,2, X. Vidal-Gómez1,2, A. Mompeón1,2, C. Hermenegildo1,2

1. Physiology, University of Valencia, Valencia, Spain. 2. INCLIVA Biomedical Research Institute, Valencia, Spain.

Estrogen vascular, and mostly endothelial, effects are meditated through signalling pathways involving both classical (ERα and ERβ) and G-protein mediated (GPER) estrogen receptors (ER). Recently, microRNA (miRNA) have emerged as key regulators of endothelial function. Since there are few studies focused on the mechanisms involving estrogen-regulation of miRNA expression in endothelial cells, we aimed to discover the role of the different estrogen receptors (ERα, ERβ and GPER) in the endothelial regulation of miRNA expression by estradiol (E2). Studies were performed in primary human umbilical vein endothelial cells (HUVEC) exposed to a physiological concentration of E2 (1 nmol/L) for 24 hours. Changes in global miRNA expression were determined by miRNA microarrays and validated by qRT-PCR. The role of different ER on the E2-modified miRNA was also analysed by qRT-PCR by using specific ER agonists (PPT for ERα, DPN for ERβ, G1 for GPER) and antagonists (ICI182780 as non-selective antagonist for ERα and ERβ, MPP for ERα and G15 for GPER). E2 modified the expression of 114 miRNA (p < 0.05): 70 miRNA were up-regulated and 44 down-regulated. To validate microarray findings, qRT-PCR was performed using another set of RNA samples. The most differentially expressed miRNA obtained in microarrays were selected and analysed by qRT-PCR: miR-30b, miR-487a, miR-4710 and miR-501 were upregulated whereas miR-378h and miR-1244 were downregulated. When studied the contribution of ER, results demonstrated ERα played a central role in the E2-modulation of miR-30b-5p, miR-378h and miR-487a-5p. In addition, E2 induction of miR-4710 was mediated through GPER, whereas miR-1244 expression depends on the three ER. E2-induced miR-501-3p expression was reverted by ICI, but not by MPP or G15, suggesting a possible role of ERβ. Unpredictably, miR-501-3p expression was not changed after isolated exposition with the different agonists. In conclusion, our results indicate that miRNA may act as crucial epigenetic regulators of gene expression in E2-treated human endothelial cells and make an important contribution on the role of ER in vascular actions mediated by E2. Moreover, they also could provide insight into the physiological mechanisms by which sex hormones levels can contribute to sex-differences in vascular function.

Where applicable, experiments conform with Society ethical requirements