The effect of various vitamin E derivatives on the urokinase-plasminogen activator system of ovine macrophages and neutrophils.
The effect of vitamin E derivatives on the urokinase-plasminogen activator (u-PA) system of resting and phorbol myristate acetate (PMA)-activated ovine macrophages and neutrophils were investigated. Blood monocyte-macrophages and neutrophils were isolated from twenty-four animals. Macrophages or neutrophils were cultured in vitro for 3 or 24 h with or without various vitamin E derivatives: free alpha-tocopherol (alpha-T), alpha-tocopheryl acetate (alpha-TA), or alpha-tocopheryl succinate (alpha-TS). Following incubation, cells were stimulated with 80 microm-PMA. Total cell-associated u-PA, membrane-bound u-PA and free u-PA binding sites were determined before and after stimulation with PMA. Results showed that none of the vitamin E derivatives had any effect (P>0.05) on the u-PA system of resting monocyte-macrophages or neutrophils. In contrast, alpha-TS, but not alpha-TA or alpha-T, increased (P<0.01) total cell-associated u-PA and membrane-bound u-PA of PMA-stimulated macrophages and neutrophils. alpha-TS had no effect (P>0.05) on total u-PA and membrane-bound u-PA activities of macrophages and neutrophils cultured in the presence of 4-phorbol 12,13 didecanoate, a phorbol ester that does not activate protein kinase (PK) C. Addition of H7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride), which is a potent inhibitor of both PK A and C, completely abolished the effect of alpha-TS on total cell-associated u-PA and membrane-bound u-PA of PMA-activated macrophages and neutrophils. Addition of HA1004 (N-(2-quanidinoethyl)-5-isoquinoline sulfonamide hydrochloride), which is a potent PK A but a weak PK C inhibitor, had no effect (P>0.05) on total cell-associated u-PA and membrane-bound u-PA of PMA-activated macrophages and neutrophils cultured in the presence of alpha-TS. Thus, PK C modulates the effect of alpha-TS on the u-PA system of ovine macrophages and neutrophils.
Politis
I
I
Department of Animal Science, Agricultural University of Athens, Athens 118 55, Greece. i.politis@aua.gr
Voudouri
A
A
Bizelis
I
I
Zervas
G
G
D000818:Animals; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D018375:Neutrophil Activation; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D012756:Sheep; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D024505:Tocopherols; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis / Q000378:metabolism*; D014810:Vitamin E / Q000031:analogs & derivatives* / Q000494:pharmacology*; D024502:alpha-Tocopherol / Q000031:analogs & derivatives* / Q000494:pharmacology
D004791:Enzyme Inhibitors; D014810:Vitamin E; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D014568:Urokinase-Type Plasminogen Activator; D024502:alpha-Tocopherol; D013755:Tetradecanoylphorbol Acetate; D024505:Tocopherols