Metabolomic responses in brain endothelial cells (CLU‐512‐hCMEC/D) to Angiotensin II and Angiotensin 1‐7

Abstract

Metabolomics is the discipline focused on the study of the dynamics, composition, interactions and responses to changes in metabolites in cells, tissues and biological fluids. Vasoactive peptides such as angiotensins cause multiple metabolic changes in vascular tissues, however these changes have not been characterized in detail. Angiotensin 1‐7 is a peptide derived from angiotensin II through the action of Angiotensin Converting Enzyme type II. This exploratory study seeks to understand the metabolic responses of a human cerebral microvascular endothelial endothelial cell line to stimulation with vasoactive peptides, in vitro (Angiotensin II, Angiotensin 1‐7). Methods We performed single cultures of the cell line (CLU‐512‐hCMEC/D), which was treated with Ang II, Ang 1‐7 or vehicle. Cells and culture medium were sampled before and after treatments. The samples were subjected to the process of extraction of metabolites with organic solvents. Each treatment was prepared in three biological replicates and was followed by metabolite extraction. Determination of metabolic profiles was performed by multiplatform, non‐targeted, metabolomic analysis, using liquid chromatography (LC) coupled to high resolution mass spectrometry (HRMS). To determine the differences in metabolic profiles between groups and to select statistically significant metabolites, multivariate statistical analysis was performed. Results We found 122 metabolites that varied between groups. In general, Ang II‐induced changes were similar to those found for Ang 1‐7. In particular, we found a reduction in the levels of several types of lipids including some bile salts, sphingolipids, glycerophospholipids and fatty acids. This study is our first step in characterizing methods to study the impact of vasoactive peptides and inflammatory molecules on the blood‐brain barrier metabolome. Studying the modulation of the vascular tissue metabolome and endothelial cells could enhance our understanding of the effect of angiotensin 1‐7 and ACE2 in pathologies such as COVID‐19.