Methods to Analyze the Effect of Diet-Derived Metabolites on Endothelial Inflammation and Cell Surface Glycosaminoglycans

Abstract

The glycocalyx is a biologically active barrier that covers the luminal side of the vascular endothelium and it is comprised of proteoglycans [core proteins with glycosaminoglycans (GAG) side chains], glycoproteins, and plasma proteins. Evidence shows that the disruption in the structure and function of the endothelial glycocalyx exacerbates vascular inflammation and atherosclerosis. The GAG components of the glycocalyx undergo remodeling in the setting of diabetes and these alterations in endothelial GAGs negatively impact the vascular function. Hence, the preservation and restoration of GAGs in altered vasculature may be a novel strategy to ameliorate vascular complications in diabetes and metabolic syndrome. Human studies support the beneficial vascular effects of flavonoids which are widely found in fruits and vegetables. Flavonoids are extensively metabolized by the intestinal microbiota and digestive enzymes in humans, suggesting that their biological activities may be mediated by their circulating metabolites. Studies indicate that counteracting the damage to GAGs using dietary compounds improve vascular complications. In this article, we describe the methods to analyze the effect of diet-derived metabolites such as metabolites of flavonoids on endothelial inflammation and cell surface glycosaminoglycans.