Antioxidant regulation of phorbol ester-induced adhesion of human Jurkat T-cells to endothelial cells.
Regulation of adhesion molecule expression and function by reactive oxygen species via specific redox sensitive mechanisms have been reported. The effects of clinically safe antioxidants in the regulation of adhesion molecule expression in human endothelial cells (ECV), and adherence of human Jurkat T cells to ECV cells were investigated. The thiol antioxidant, alpha-lipoate, at clinically relevant doses down-regulated phorbol 12-myristate 13-acetate (PMA)-induced adhesion molecule expression and cell-cell adhesion. Inhibition of PMA-induced ICAM-1 and VCAM-1 expression as well as PMA-induced adhesion of Jurkat T-cells to ECV cells by alpha-lipoate was dose dependent (50-250 microM). The effect was significant for ICAM-1 (p < .01) and VCAM-1 (p < .01) expression in cells pretreated with 100 microM alpha-lipoate compared to PMA-activated untreated cells. Inhibition of PMA-induced adhesion molecule expression and cell-cell adhesion was more pronounced when a combination of antioxidants, alpha-lipoate and alpha-tocopherol, were used compared to the use of either of these antioxidant alone. The regulation of adhesion molecule expression and function by low concentration of antioxidants investigated does not appear to be NF-kappaB regulated or transcription dependent because no change in the mRNA response was observed. Protein kinase C (PKC) has been suggested to regulate PMA-induced adhesion molecule expression by post-transcriptional stabilization of adhesion molecule mRNA. Alpha-lipoate pretreatment did not influence the response of PKC activity to PMA. Oxidants are known to be involved in the regulation of cell adhesion processes. Treatment of ECV cells with PMA induced generation of intracellular oxidants. Alpha-lipoate (100 or 250 microM) treatment decreased PMA-induced generation of intracellular oxidants. The inhibitory effect of low concentration of alpha-lipaote alone or in combination with alpha-tocopherol on agonist-induced adhesion processes observed in this study may be of potential therapeutic value.
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Department of Molecular and Cell Biology, University of California, Berkeley 94720-3200, USA. sashwati@socrates.berkeley.edu
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D000975:Antioxidants / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects; D002461:Cell Line, Transformed; D015536:Down-Regulation / Q000187:drug effects; D004357:Drug Synergism; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D007424:Intracellular Fluid / Q000737:chemistry / Q000187:drug effects; D019169:Jurkat Cells; D016328:NF-kappa B / Q000187:drug effects / Q000378:metabolism; D010545:Peroxides / Q000032:analysis; D011493:Protein Kinase C / Q000187:drug effects / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis / Q000187:drug effects; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects*; D013755:Tetradecanoylphorbol Acetate / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D008063:Thioctic Acid / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics; D014810:Vitamin E / Q000494:pharmacology
D000975:Antioxidants; D016328:NF-kappa B; D010545:Peroxides; D012333:RNA, Messenger; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D014810:Vitamin E; D008063:Thioctic Acid; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate