Effects of resveratrol on CYP450-derived arachidonic acid metabolites and mRNA gene expression in human endothelial HUVEC cells.

Abstract

Objectives: This study aims to investigate the expression of arachidonic acid (ARA)-metabolizing cytochrome P450 (CYP450) enzymes and their metabolites, as well as the effects of resveratrol, a compound known for its cardiovascular benefits, on these enzymes in human umbilical vein endothelial cells (HUVECs).

Methods: HUVECs were cultured and treated with 10 µM resveratrol for 24 h. The mRNA expression of key ARA-metabolizing CYP450 enzymes was analyzed using real-time polymerase chain reaction. ARA metabolites produced by CYP450s were identified and quantified in cell culture media and lysates using liquid chromatography coupled with mass spectrometry. In addition, in silico docking was done to evaluate the effect of resveratrol on the identified CYP450s in HUVECS.

Results: CYP2J2, CYP2U1, and epoxide hydrolase were expressed in HUVECs; however, their mRNA expression levels were not significantly altered following resveratrol treatment. The metabolite 20-hydroxyeicosatetraenoic acid (20-HETE) and its synthesizing CYP450 genes were not detected or below the lower limit of detection in HUVECs. Additionally, several epoxyeicosatrienoic acids (EETs) and dihydroxyeicosatrienoic acids (DHETEs) were present in significantly higher amounts in the cell culture media compared to the cell lysates. Resveratrol treatment significantly reduced (p<0.05, t-test) the levels of EETs and DHETEs. In addition, in silico analysis showed resveratrol has a moderate affinity to interact with CYP2J2, the major EETs producer.

Conclusions: These findings suggest that resveratrol significantly reduces the levels of EETs and DHETEs possibly through direct interaction with CYP2J2. Furthermore, HUVECs may not be a suitable in vitro model for studying ARA metabolism to 20-HETE.