EFFECT OF THE STIMULATION WITH (-) EPICATECHIN ON GENE EXPRESSION IN HUVEC CELLS

Background and objectives: Epicatechin (EC) is the most abundant polyphenol in cocoa. Consumption of cocoa polyphenols has been related with cardiovascular benefits such as decrease in blood pressure and improvement of markers of glucose and lipid metabolism in human. These polyphenols have been shown to modulate gene expression and exert an anti-inflammatory effect. The present study analyzed the dose-response effect of EC stimulation on the expression of genes related with inflammation (NFkB) and endothelial cell function (eNOS) in primary culture of human umbilical vein endothelial cells (HUVEC). The tested doses included concentrations that are achievable by intake of EC containing food. Methods: HUVEC cells were isolated from the umbilical cord of women with a history of healthy pregnancy who signed the informed consent. Endothelial cells were isolated and cultured using standard methods. Cells were stimulated with three different doses of EC (1, 0.1, 0.01 uM and untreated control) during 4 h. All experiments were conducted in triplicates. RNA was isolated from cells and eNOS and NFkB (p50) mRNA was measured using real time, quantitative RT-PCR by absolute quantitation. HUVEC cells were identified by immunoflorescence staining. Comparison of the mRNA abundance across treatments was conducted by a one-way ANOVA. Statistical analysis was conducted in SPSS v.20 software. This project was approved by the Ethics committee at INMEGEN. Results: The immunoflorescence staining was positive for endothelial cells. Significant differences in mRNA abundance were found for the analyzed genes. Stimulation with EC at 0.01 uM decreased the expression of eNOS mRNA as compared with control (p