Human endothelial cells are targets for platelet-activating factor (PAF). Activation of alpha and beta protein kinase C isozymes in endothelial cells stimulated by PAF.
We evaluated the role of the protein kinase C (PKC) and its isozymes in the activation of human endothelial cells (EC) stimulated by platelet-activating factor (PAF). Exposure of confluent EC to PAF resulted in a rapid and concentration-dependent redistribution of PKC from cytosol to plasma-membrane, rearrangement of cytoskeleton (i.e. decrease in F-actin content and redistribution of vinculin), and finally increase in the transendothelial flux of 125I-albumin. Stimulation of EC with oleylacetylglycerol or phorbol 12-myristate 13-acetate induced the modification of the cytoskeletal structures and the increase of 125I-albumin clearance. Inhibitors of PKC prevented the effects induced by PAF on the cytoskeleton and on the barrier function of the EC monolayer. Confluent EC expressed only alpha, beta, and epsilon PKC isoforms. Biochemical and immunochemical analysis showed that the time course of the PKC isozymes translocation from cytosol to the membrane fraction of EC stimulated by PAF was different: beta isoform was redistributed more quickly than alpha isoform. PAF did not induce translocation of PKC epsilon. These results suggest that activation of PKC alpha and beta is an important signal transduction pathway by which PAF activates endothelial monolayer and modify its function of barrier to macromolecules.
Bussolino
F
F
Department of Genetics, Biology and Medical Chemistry, University of Torino, Italy.
Silvagno
F
F
Garbarino
G
G
Costamagna
C
C
Sanavio
F
F
Arese
M
M
Soldi
R
R
Aglietta
M
M
Pescarmona
G
G
Camussi
G
G
D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D000906:Antibodies; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D004075:Diglycerides / Q000494:pharmacology; D004224:Diterpenes; D004730:Endothelium, Vascular / Q000187:drug effects / Q000201:enzymology*; D004789:Enzyme Activation; D046934:Ginkgolides; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D007700:Kinetics; D007783:Lactones / Q000494:pharmacology; D008969:Molecular Sequence Data; D009281:Naphthalenes; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D010742:Phospholipid Ethers / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D011083:Polycyclic Compounds / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D047428:Protein Kinase Inhibitors; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D016596:Vinculin / Q000378:metabolism
D000199:Actins; D000906:Antibodies; D003598:Cytoskeletal Proteins; D004075:Diglycerides; D004224:Diterpenes; D046934:Ginkgolides; D007527:Isoenzymes; D007783:Lactones; D009281:Naphthalenes; C029271:O-deacetyl platelet activating factor; D010455:Peptides; D010742:Phospholipid Ethers; D010972:Platelet Activating Factor; D011083:Polycyclic Compounds; D047428:Protein Kinase Inhibitors; D013844:Thiazoles; C049820:SRI 63072; D016596:Vinculin; C037913:CV 3988; C038237:1-oleoyl-2-acetylglycerol; C045856:ginkgolide B; D011493:Protein Kinase C; C058819:calphostin C; D013755:Tetradecanoylphorbol Acetate