Structural alteration of the endothelial glycocalyx: contribution of the actin cytoskeleton

The endothelial glycocalyx is a carbohydrate–protein layer that lines the luminal surface of the endothelium. It anchors to the cell membrane via its core proteins that share extended link to the actin cytoskeleton. It is widely accepted that those protein domains and the attached carbohydrates are susceptible to pathological changes. It is unclear, however, to what extent the actin cytoskeleton contributes to the glycocalyx stability. In this study, we investigate the role of the actin cytoskeleton in the maintenance of the glycocalyx under static and laminar flow conditions in vitro. Our results show that in the static culture medium neither rapid actin depolymerisation nor prolonged actin disturbance leads to glycocalyx disruption from the apical surface of human umbilical vein endothelial cells. However, when endothelial cells are exposed to laminar flow for 24 h, the glycocalyx is seen to shift to the downstream peripheral region of the cell surface. The mean fluorescence intensity decreases to (91.9 pm 2.5%) of the control. When actin depolymerisation is introduced, the intensity decreases significantly to (54.7 pm 1.3%), indicating a severe disruption of the glycocalyx. Similar changes are observed in human aortic endothelial cells, where the intensity of the glycocalyx is reduced to (72.8 pm 1.6%) of the control. Collectively, we demonstrate that the actin cytoskeleton contributes to structural stability of the glycocalyx under shear stress. Our results can be used to develop new strategies to prevent shedding of the glycocalyx in cardiovascular diseases.