Tumor necrosis factor-alpha, platelet-activating factor, and hydrogen peroxide activate protein kinase C subtypes alpha and epsilon in human saphenous vein endothelial cells.
Protein kinase C (PKC), the major receptor for tumor-promoting phorbol esters, consists of a family of at least 12 distinct lipid-regulated enzymes. We examined the expression and regulation of PKC isoforms in human saphenous vein endothelial cells (HSVEC). Western blot analysis with PKC isoform-specific antibodies indicated that PKC alpha, PKC epsilon and PKC zeta were expressed in these cells. Translocation and down-regulation of PKC alpha and epsilon but not zeta were detected by short-term and long-term treatment with TPA (12-O-tetradecanoylphorbol 13-acetate), respectively. Tumor necrosis factor-alpha (TNF-alpha 1,600 U/ml) and platelet activating factor (PAF 50 nM) increased the membrane content of PKC alpha and epsilon but not zeta. H2O2 (10 mM) induced the translocation of PKC alpha from the cytosol to the membrane and increased PKC epsilon content in both cytosol and membrane. However, 12-(S)-HETE (12-hydroxyeicosatetraenoic acid) (100 nM), a lipoxygenase metabolite of arachidonic acid, did not affect the two isoforms. These results suggest that the molecular action of TNF-alpha, PAF, and H2O2 in HSVEC might occur through PKC alpha and epsilon activation.
Chen
C C
CC
Department of Pharmacology and Internal Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan.
Liau
C S
CS
Lee
Y T
YT
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism; D015153:Blotting, Western; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000494:pharmacology*; D011493:Protein Kinase C / Q000096:biosynthesis / Q000378:metabolism*; D012501:Saphenous Vein / Q000166:cytology / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D007527:Isoenzymes; D010972:Platelet Activating Factor; D014409:Tumor Necrosis Factor-alpha; D016718:Arachidonic Acid; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D006861:Hydrogen Peroxide; D011493:Protein Kinase C