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Ceramide mediates inhibition of the Akt/eNOS pathway by high levels of glucose in human vascular endothelial cells
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Aimin Wang
Published/Copyright:
November 9, 2012
Abstract
Objective: 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).
Materials and methods: 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.
Results: High-glucose levels induce ceramide accumulation in a dose- and time-dependent manner (p<0.05). We also show that exposure of HUVECs to high-glucose conditions inhibits the insulin-mediated activation of Akt/eNOS signalling and the subsequent NO generation in a dose-dependent manner (p<0.05). Preventing de novo ceramide synthesis attenuated the antagonistic effects of high-glucose levels on the Akt/eNOS signalling pathway (p<0.05); conversely, inducing ceramide build-up augmented the inhibitory effects of high-glucose levels on the Akt/eNOS signalling pathway (p<0.05).
Conclusion: Ceramide is both necessary and sufficient for mediating the inhibition of the Akt/eNOS signalling pathway by high-glucose levels in endothelial cells.
Keywords: Akt/endothelial nitric oxide synthase (eNOS); ceramide; glucose; nitric oxide (NO); type 2 diabetes mellitus
Received: 2012-5-4
Accepted: 2012-9-21
Published Online: 2012-11-09
Published in Print: 2013-02-01
©2013 by Walter de Gruyter Berlin Boston
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Keywords for this article
Akt/endothelial nitric oxide synthase (eNOS);
ceramide;
glucose;
nitric oxide (NO);
type 2 diabetes mellitus
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