To specify the site of action of the synthetic coumarin derivatives 7-hydroxy-3-(4'-hydroxyphenyl) coumarin (HHC) and 7-hydroxy-3-(4'-hydroxyphenyl) dihydrocoumarin (HHDC), we evaluated their effects on extra- and intracellular reactive oxygen species (ROS) formation in phorbol-myristate-13-acetate (PMA) stimulated human neutrophils. We studied also the effects of HHC and HHDC on possible molecular mechanisms which participate in the activation of NADPH oxidase, that is, on PKC activity, on phosphorylation of some PKC isoforms (α, βII, and δ), and on phosphorylation of the NADPH oxidase subunit p40(phox). Without affecting cytotoxicity, both coumarines tested were effective inhibitors/scavengers of ROS produced by neutrophils on extracellular level. HHC markedly diminished oxidant production and also, intracellularly, decreased PKC activity and partly phosphorylation of PKCα, βII. On the other hand, we did not observe any effect of coumarin derivatives on phosphorylation of PKC δ and on phosphorylation of the NADPH oxidase subunit p40(phox), which were suggested to be involved in the PMA-dependent intracellular activation process. In agreement with our previous findings, we assume that the different molecular structures of HHC and HHDC with their different physicochemical and free radical scavenging characteristics are responsible for their diverse effects on the parameters tested.

Institute of Chemical Technology Prague Technická 5 166 28 Prague Czech Republic

Institute of Experimental Pharmacology and Toxicology Slovak Academy of Sciences Dúbravská cesta 9 841 04 Bratislava Slovakia

Institute of Organic Chemistry and Biochemistry Academy of Sciences of the Czech Republic v v i Flemingovo náměsti 2 161 10 Prague Czech Republic

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Riveiro ME, Moglioni A, Vazquez R, et al. Structural insights into hydroxycoumarin-induced apoptosis in U-937 cells. Bioorganic and Medicinal Chemistry. 2008;16(5):2665–2675. PubMed

Belluti F, Fontana G, Bo LD, Carenini N, Giommarelli C, Zunino F. Design, synthesis and anticancer activities of stilbene-coumarin hybrid compounds: identification of novel proapoptotic agents. Bioorganic and Medicinal Chemistry. 2010;18(10):3543–3550. PubMed

Kostova I, Bhatia S, Grigorov P, et al. Coumarins as antioxidants. Current Medicinal Chemistry. 2011;18(25):3929–3951. PubMed

Sashidhara KV, Kumar A, Dodda RP, et al. Coumarin-trioxane hybrids: synthesis and evaluation as a new class of antimalarial scaffolds. Bioorganic & Medicinal Chemistry Letters. 2012;22(12):3926–3930. PubMed

Ramasamy R, Maqbool M, Mohamed AL, Noah RM. Elevated neutrophil respiratory burst activity in essential hypertensive patients. Cellular Immunology. 2010;263(2):230–234. PubMed

Ciz M, Denev P, Kratchanova M, Vasicek O, Ambrozova G, Lojek A. Flavonoids inhibit the respiratory burst of neutrophils in mammals. Oxidative Medicine and Cellular Longevity. 2012;2012:6 pages.181295 PubMed PMC

Khlebnikov AI, Schepetkin IA, Domina NG, Kirpotina LN, Quinn MT. Improved quantitative structure-activity relationship models to predict antioxidant activity of flavonoids in chemical, enzymatic, and cellular systems. Bioorganic and Medicinal Chemistry. 2007;15(4):1749–1770. PubMed PMC

Gauss KA, Nelson-Overton LK, Siemsen DW, Gao Y, DeLeo FR, Quinn MT. Role of NF-κB in transcriptional regulation of the phagocyte NADPH oxidase by tumor necrosis factor-α . Journal of Leukocyte Biology. 2007;82(3):729–741. PubMed

Raad H, Paclet MH, Boussetta T, et al. Regulation of the phagocyte NADPH oxidase activity: phosphorylation of gp91phox/NOX2 by protein kinase C enhances its diaphorase activity and binding to Rac2, p67phox, and p47phox. FASEB Journal. 2009;23(4):1011–1022. PubMed PMC

Bylund J, Brown KL, Movitz C, Dahlgren C, Karlsson A. Intracellular generation of superoxide by the phagocyte NADPH oxidase: how, where, and what for? Free Radical Biology and Medicine. 2010;49(12):1834–1845. PubMed

Karlsson A, Dahlgren C. Assembly and activation of the neutrophil NADPH oxidase in granule membranes. Antioxidants and Redox Signaling. 2002;4(1):49–60. PubMed

Matute JD, Arias AA, Wright NAM, et al. A new genetic subgroup of chronic granulomatous disease with autosomal recessive mutations in p40phox and selective defects in neutrophil NADPH oxidase activity. Blood. 2009;114(15):3309–3315. PubMed PMC

Björnsdottir H, Granfeldt D, Welin A, Bylund J, Karlsson A. Inhibition of phospholipase A2 abrogates intracellular processing of NADPH-oxidase derived reactive oxygen species in human neutrophils. Experimental Cell Research. 2013;319(5):761–774. PubMed

Nimeri G, Majeed M, Elwing H, Öhman L, Wetterö J, Bengtsson T. Oxygen radical production in neutrophils interacting with platelets and surface-immobilized plasma proteins: role of tyrosine phosphorylation. Journal of Biomedical Materials Research A. 2003;67(2):439–447. PubMed

El-Benna J, Dang PMC, Gougerot-Pocidalo MA, Marie JC, Braut-Boucher F. p47phox, the phagocyte NADPH oxidase/NOX2 organizer: structure, phosphorylation and implication in diseases. Experimental and Molecular Medicine. 2009;41(4):217–225. PubMed PMC

Hong NJ, Silva GB, Garvin JL. PKC-α mediates flow-stimulated superoxide production in thick ascending limbs. American Journal of Physiology—Renal Physiology. 2010;298(4):F885–F891. PubMed PMC

Korchak HM, Dorsey LB, Li H, Mackie D, Kilpatrick LE. Selective roles for α-PKC in positive signaling for O2-generation and calcium mobilization but not elastase release in differentiated HL60 cells. Biochimica et Biophysica Acta. 2007;1773(3):440–449. PubMed

Nixon JB, McPhail LC. Protein kinase C (PKC) isoforms translocate to triton-insoluble fractions in stimulated human neutrophils: correlation of conventional PKC with activation of NADPH oxidase. Journal of Immunology. 1999;163(8):4574–4582. PubMed

Bertram A, Ley K. Protein kinase C isoforms in neutrophil adhesion and activation. Archivum Immunologiae et Therapiae Experimentalis. 2011;59(2):79–87. PubMed

Vrba J, Dvořák Z, Ulrichová J, Modrianský M. Conventional protein kinase C isoenzymes undergo dephosphorylation in neutrophil-like HL-60 cells treated by chelerythrine or sanguinarine. Cell Biology and Toxicology. 2008;24(1):39–53. PubMed

Kilpatrick LE, Sun S, Li H, Vary TC, Korchak HM. Regulation of TNF-induced oxygen radical production in human neutrophils: role of δ-PKC. Journal of Leukocyte Biology. 2010;87(1):153–164. PubMed PMC

Brown GE, Stewart MQ, Liu H, Ha VL, Yaffe MB. A novel assay system implicates PtdIns(3,4)P2, PtdIns(3)P, and PKCδ in intracellular production of reactive oxygen species by the NADPH oxidase. Molecular Cell. 2003;11(1):35–47. PubMed

Drábiková K, Perečko T, Nosáľ R, et al. Different effect of two synthetic coumarin-stilbene hybrid compounds on phagocyte activity. Neuroendocrinology Letters. 2010;31:73–78. PubMed

Jančinová V, Perečko T, Nosáľ R, Košťálová D, Bauerová K, Drábiková K. Decreased activity of neutrophils in the presence of diferuloylmethane (curcumin) involves protein kinase C inhibition. European Journal of Pharmacology. 2009;612:161–166. PubMed

Nosáľ R, Perečko T, Jančinová V, Drábiková K, Harmatha J, Sviteková K. Naturally appearing N-feruloylserotonin isomers suppress oxidative burst of human neutrophils at the protein kinase C level. Pharmacological Reports. 2011;63:790–798. PubMed

Drábiková K, Perečko T, Nosál R, et al. Glucomannan reduces neutrophil free radical production in vitro and in rats with adjuvant arthritis. Pharmacological Research. 2009;59:399–403. PubMed

Jančinová V, Perečko T, Nosáľ R, Harmatha J, Šmidrkal J, Drábiková K. The natural stilbenoid pinosylvin and activated neutrophils: effects on oxidative burst, protein kinase C, apoptosis and efficiency in adjuvant arthritis. Acta Pharmacologica Sinica. 2012;33:1285–1292. PubMed PMC

Drábiková K, Jančinová V, Nosáľ R, Pečivová J, Mačičková T, Turčáni P. Inibitory effect of stobadine on FMLP-induced chemiluminescence in human whole blood and isolated polymorphonuclear leukocytes. Luminescence. 2007;22:67–71. PubMed

Varga Z, Kosaras E, Komodi E, et al. Effects of tocopherols and 2,2′-carboxyethyl hydroxychromans on phorbol-ester-stimulated neutrophils. Journal of Nutritional Biochemistry. 2008;19(5):320–327. PubMed

Stevenson DE, Hurst RD. Polyphenolic phytochemicals—just antioxidants or much more? Cellular and Molecular Life Sciences. 2007;64(22):2900–2916. PubMed PMC

Pandey KB, Rizvi SI. Plant polyphenols as dietary antioxidants in human health and disease. Oxidative Medicine and Cellular Longevity. 2009;2(5):270–278. PubMed PMC

Obrenovich ME, Nair NG, Beyaz A, Aliev G, Reddy VP. The role of polyphenolic antioxidants in health, disease, and aging. Rejuvenation Research. 2010;13(6):631–643. PubMed

Štefek M. Natural flavonoids as potential multifunctional agents in prevention of diabetic cataract. Interdisciplinary Toxicology. 2011;4(2):69–77. PubMed PMC

Perečko T, Jančinová V, Drábiková K, Nosál’ R, Harmatha J. Structure-efficiency relationship in derivatives of stilbene. Comparison of resveratrol, pinosylvin and pterostilbene. Neuroendocrinology Letters. 2008;29(5):802–805. PubMed

Perečko T, Drábiková K, Nosáľ R, Harmatha J, Jančinová V. Pharmacological modulation of activated neutrophils by natural polyphenols. In: Pandalai SG, editor. Recent Research Developments in Pharmacology. Vol. 2. Trivandrum, India: Research Signpost; 2011. pp. 27–67.

Dahlgren C, Karlsson A. Respiratory burst in human neutrophils. Journal of Immunological Methods. 1999;232(1-2):3–14. PubMed

Kabeya LM, de Marchi AA, Kanashiro A, et al. Inhibition of horseradish peroxidase catalytic activity by new 3-phenylcoumarin derivatives: synthesis and structure-activity relationships. Bioorganic and Medicinal Chemistry. 2007;15(3):1516–1524. PubMed

Andrade MF, Kabeya LM, Azzolini AE, et al. 3-Phenylcoumarin derivatives selectively modulate different steps of reactive oxygen species production by immune complex-stimulated human neutrophils. International Immunopharmacology. 2013;1:387–394. PubMed

Pečivová J, Mačičková T, Harmatha J, Sviteková K, Nosáľ R. In vitro effect of pinosylvin and pterostilbene on human neutrophils. Interdisciplinary Toxicology. 2010;3(article A73)

Varga Z, Seres I, Nagy E, et al. Structure prerequisite for antioxidant activity of silybin in different biochemical systems in vitro. Phytomedicine. 2006;13(1-2):85–93. PubMed

Lin HC, Tsai SH, Chen CS, et al. Structure-activity relationship of coumarin derivatives on xanthine oxidase-inhibiting and free radical-scavenging activities. Biochemical Pharmacology. 2008;75(6):1416–1425. PubMed

Devienne KF, Cálgaro-Helena AF, Dorta DJ, et al. Antioxidant activity of isocoumarins isolated from Paepalanthus bromelioides on mitochondria. Phytochemistry. 2007;68(7):1075–1080. PubMed

Fraga CG, Galleano M, Verstraeten SV, Oteiza PI. Basic biochemical mechanisms behind the health benefits of polyphenols. Molecular Aspects of Medicine. 2010;31(6):435–445. PubMed

Nosáľ R, Perečko T, Jančinová V, Drábiková K, Harmatha J, Sviteková K. Suppression of oxidative burst in human neutrophils with the naturally occurring serotonin derivative isomer from Leuzea carthamoides. Neuroendocrinology Letters. 2010;31:69–72. PubMed

Jančinová V, Perečko T, Drábiková K, Nosáľ R, Sviteková K. Piceatannol, a natural analogue of resveratrol, inhibits oxidative burst of human neutrophils. Interdisciplinary Toxicology. 2011;4:A35–A36.

Bouin AP, Grandvaux N, Vignais PV, Fuchs A. p40(phox) is phosphorylated on threonine 154 and serine 315 during activation of the phagocyte NADPH oxidase: implication of a protein kinase C-type kinase in the phosphorylation process. Journal of Biological Chemistry. 1998;273(46):30097–30103. PubMed

Someya A, Nunoi H, Hasebe T, Nagaoka I. Phosphorylation of p40-phox during activation of neutrophil NADPH oxidase. Journal of Leukocyte Biology. 1999;66(5):851–857. PubMed