High glucose concentrations increase endothelial cell permeability via activation of protein kinase C alpha

Item Type:	Article
Title:	High glucose concentrations increase endothelial cell permeability via activation of protein kinase C alpha
Creators Name:	Hempel, A. and Maasch, C. and Heintze, U. and Lindschau, C. and Dietz, R. and Luft, F.C. and Haller, H.
Abstract:	Endothelial cell permeability is impaired in diabetes mellitus and may be increased by high extracellular glucose concentrations. High glucose activates protein kinase C (PKC), a family of kinases vital to intracellular signaling. We tested the hypothesis that high glucose concentration activates PKC in endothelial cells and leads to an increase in endothelial cell permeability via distinct PKC isoforms. Porcine aortic endothelial cells were used, and the PKC isoforms alpha, delta, epsilon, zeta, and theta were identified in these cells. Glucose caused a rapid dose-dependent increase in endothelial cell permeability, with an EC50 of 17.5 mmol/L. Phorbol 12-myristate 13-acetate (TPA) induced an increase in permeability very similar to that elicited by glucose. The effect of glucose and TPA was totally reversed by preincubating the cells with the PKC inhibitors staurosporine (10(-8) mol/L) and Goe 6976 (10(-8) mol/L). Downregulation of PKC by preincubation with TPA for 24 hours also abolished the effect of glucose and TPA on endothelial cell permeability. High glucose (20 mmol/L) caused an increase in PKC activity at 2, 10, and 30 minutes. Cell fractionation and Western blot analysis showed a glucose-induced translocation of PKC alpha and PKC epsilon. Confocal microscopy confirmed the translocation and showed an association of PKC alpha and PKC epsilon with nuclear structures and the cell membrane. Specific antisense oligodesoxynucleotides (ODNs) against PKC alpha reduced the expression of the isoform, abolished the effects of glucose on endothelial cell permeability completely, and reduced the TPA effect significantly. In contrast, specific antisense ODNs against PKC epsilon had no effect on glucose-induced permeability and only a minor effect on the TPA-induced increase in permeability. We conclude that an increase in extracellular glucose leads to a rapid dose-dependent increase in endothelial cell permeability via the activiation of PKC and that this effect is mediated by the PKC isoform alpha.
Keywords:	Permeability, Glucose, Hyperglycemia, Protein Kinase C, Antisense Oligodesoxynucleotide, Animals, Swine
Source:	Circulation Research
ISSN:	0009-7330
Publisher:	American Heart Association
Volume:	81
Number:	3
Page Range:	363-371
Date:	September 1997
Official Publication:	http://circres.ahajournals.org/cgi/content/abstract/81/3/363
PubMed:	View item in PubMed

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