Ceramide mediates inhibition of the Akt/eNOS pathway by high levels of glucose in human vascular endothelial cells

Abstract To investigate how ceramide mediates the effects of high-glucose-induced inhibition of the Akt/endothelial nitric oxide synthase (eNOS) signalling pathway in human vascular endothelial cells (HUVECs). NO levels were determined by ELISA. Endogenous ceramide levels were determined using a liquid chromatography-mass spectrometry assay. Akt and eNOS protein expressions were determined by Western blotting. High-glucose levels induce ceramide accumulation in a dose- and time-dependent manner (p