IMPAIRED MEMBRANE TRANSLOCATION OF THROMBIN-STIMULATED PKC-EPSILON BYHIGH GLUCOSE

Hyperglycaemia is known to cause endothelial dysfunction and to promot e diabetic angiopathy. Therefore, this study was designed to evaluate the effect of long term incubation (16+/-1 days) in 30 mM vs 5 mM gluc ose on ligand induced translocation of protein kinase C (PKC) in paire d cultures of individual isolates of human umbilical vein endothelial cells (HUVECs). Cells were stimulated with increasing concentrations o f thrombin (0.01, 0.1, 1, 10 and 100 nM) for 30 seconds in the presenc e of 5 mM and 30 mM glucose, respectively, and analyzed by immunoblott ing for PKC-isoforms alpha and epsilon. Stimulation by thrombin of con fluent cultures displayed a concentration dependent rise in membrane b ound PKC alpha and epsilon.