PMID: 9659299Jul 11, 1998Paper

Impaired membrane translocation of thrombin stimulated PKC epsilon by high glucose

The Journal of Endocrinology
G ReiningW Waldhäusl

Abstract

Hyperglycaemia is known to cause endothelial dysfunction and to promote diabetic angiopathy. Therefore, this study was designed to evaluate the effect of long term incubation (16 +/- 1 days) in 30 mM vs 5 mM glucose on ligand induced translocation of protein kinase C (PKC) in paired cultures of individual isolates of human umbilical vein endothelial cells (HUVECs). Cells were stimulated with increasing concentrations of thrombin (0.01, 0.1, 1, 10 and 100 nM) for 30 seconds in the presence of 5 mM and 30 mM glucose, respectively, and analyzed by immunoblotting for PKC-isoforms alpha and epsilon. Stimulation by thrombin of confluent cultures displayed a concentration dependent rise in membrane bound PKC alpha and epsilon. Translocation of PKC alpha by thrombin remained unaffected by high versus normal ambient glucose, whereas translocation of PKC epsilon in cells grown in 30mM glucose was reduced at maximal thrombin concentrations (area under the curve, AUC: 90.4 +/- 7% of control cells; p < 0.008; n = 6) versus control cultures kept in 5mM glucose. In the identical isolates translocation of PKC epsilon was not reduced by 30 mM mannitol used as osmotic control. No change was induced by long term incubation of resting cells with 3...Continue Reading

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