Innovative assessment of lipid‐induced oxidative stress and inflammation in harvested human endothelial cells

Abstract

Abstract Studying acute changes in vascular endothelial cells in humans is challenging. We studied ten African American women and used the J‐wire technique to isolate vein endothelial cells before and after a four‐hour lipid and heparin infusion. Dynamic changes in lipid‐induced oxidative stress and inflammatory markers were measured with fluorescence‐activated cell sorting. We used the surface markers CD31 and CD144 to identify human endothelial cells. Peripheral blood mononuclear cells isolated from blood were used as a negative control. The participants received galantamine (16 mg/day) for 3 months. We previously demonstrated that galantamine treatment effectively suppresses lipid‐induced oxidative stress and inflammation. In this study, we infused lipids to evaluate its potential to increase the activation of endothelial cells, as assessed by the levels of CD54+ endothelial cells and expression of Growth arrest‐specific 6 compared to the baseline sample. Further, we aimed to investigate whether lipid infusion led to increased expression of the oxidative stress markers IsoLGs and nitrotyrosine in endothelial cells. This approach will expedite the in vivo identification of novel pathways linked with endothelial cell dysfunction induced by oxidative stress and inflammatory cytokines. This study describes an innovative method to harvest and study human endothelial cells and demonstrates the dynamic changes in oxidative stress and inflammatory markers release induced by lipid infusion.

Keywords

• flow cytometry
• inflammation
• J‐wire technique; endothelial cells
• lipid
• oxidative stress