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Drug Res (Stuttg) 2015; 65(05): 244-251
DOI: 10.1055/s-0034-1377002
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

Diclofenac-Choline Antioxidant Activity Investigated by means of Luminol Amplified Chemiluminescence of Human Neutrophil Bursts and Electron Paramagnetic Resonance Spectroscopy

P. C. Braga
1   Department of Medical Biotechnology and Translational Medicine, School of Medicine, University of Milan, Milan, Italy
,
N. Lattuada
1   Department of Medical Biotechnology and Translational Medicine, School of Medicine, University of Milan, Milan, Italy
,
V. Greco
1   Department of Medical Biotechnology and Translational Medicine, School of Medicine, University of Milan, Milan, Italy
,
V. Sibilia
1   Department of Medical Biotechnology and Translational Medicine, School of Medicine, University of Milan, Milan, Italy
,
M. Falchi
1   Department of Medical Biotechnology and Translational Medicine, School of Medicine, University of Milan, Milan, Italy
,
T. Bianchi
2   AVIS Comunale di Milano, Ospedale Niguarda, Milano, Italy
,
M. Dal Sasso
1   Department of Medical Biotechnology and Translational Medicine, School of Medicine, University of Milan, Milan, Italy
› Author Affiliations
Further Information

Publication History

received 11 November 2013

accepted 03 May 2014

Publication Date:
11 June 2014 (online)

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Abstract

A new diclofenac salt called diclofenac-choline (DC) has recently been proposed for the symptomatic treatment of oropharyngeal inflammatory processes and pain because its greater water solubility allows the use of high concentrations, which are useful when the contact time between the drug and the oropharyngeal mucosa is brief, as in the case of mouthwashes or spray formulations. The antioxidant activity of DC has not yet been investigated, and so the aim was to use luminol-amplified-chemiluminescence (LACL) to verify whether various concentrations of DC (1.48, 0.74 and 0.37 mg/mL for incubation times of 2, 4 and 8 min) interfere with oxygen and nitrogen radicals during the course of human neutrophils respiratory bursts; electron paramagnetic resonance (EPR) spectroscopy was used to investigate its direct antiradical (scavenger) activity. The EPR findings showed that DC has concentration-dependent scavenging activity against the ABTS, the DPPH, and the hydroxyl radicals, but no activity on superoxide anion, as has been previously reported in the case of other NSAIDs. LACL revealed an inhibitory effect that was statistically significant after only 2 min of incubation, and similar after 4 and 8 min. The effects on the peroxynitrite radical paralleled those observed in the previous test. High concentrations and short incubation times showed that there is no interference on PMN viability, and so the inhibitory findings must be attributed to the effect of the drug. The anti-inflammatory effects of DC cannot be attributed solely to the inhibition of prostaglandin synthesis, but its effects on free radicals and neutrophil bursts suggest that they may contribute to its final therapeutic effect.

Key words

diclofenac-choline - electron paramagnetic resonance (EPR) - human neutrophils - chemiluminescence - antioxidant activity
 

  • References

  • 1 Rahaman I, Mac Nee W. Oxidative stress and regulation of glutathione in lung inflammation. Eur Respir J 2000; 16: 534-554
    CrossrefPubMedGoogle Scholar
  • 2 Halliwell B. Reactive oxygen species in living systems: source, biochemistry and role in human disease. Am J Med 1991; 91 3C 14-22
    PubMedGoogle Scholar
  • 3 Dallegri F, Ottonello L. Tissue injury in neutrophilic inflammation. Inflamm Res 1997; 46: 382-391
    CrossrefPubMedGoogle Scholar
  • 4 Pettersen CA, Adler K. Airways inflammation and COPD: Epithelial-neutrophil interactions. Chest 2002; 121: 142S-150S
    CrossrefPubMedGoogle Scholar
  • 5 Van Wetering S, Mannesse-Lazeroms SP, Van Sterkenburg MA et al. Neutrophil defensins stimulate the release of cytokines by airway epithelial cells: modulation bydexametasone. Inflamm Res 2002; 51: 8-15
    CrossrefPubMedGoogle Scholar
  • 6 Cassatella MA. The production of cytokine4s by polymorphonuclear neutrophils. Immunol Today 1995; 16: 21-26
    CrossrefPubMedGoogle Scholar
  • 7 Ong CKS, Seymour RA. An evidence based update of the use of analgesics in dentistry. Periodontology 2008; 46: 143-164
    CrossrefPubMedGoogle Scholar
  • 8 Dionne RA, Berthold CW. Therapeutic use of non-steroidal anti-inflammatory drugs in dentistry. Crit Rev Oral Biol Med 2001; 12: 315-330
    CrossrefPubMedGoogle Scholar
  • 9 Passali D, Volontè M, Passali GC et al. Efficacy and safety of ketoprofen lysine salt mouthwash versus benzydamine hydrochloride mouthwash in acute pharyngeal inflammation: a randomized, single-blind study. Clin Therap 2001; 23: 1508-1518
    CrossrefPubMedGoogle Scholar
  • 10 Gan TJ. Diclofenac: un update on its mechanism of action and safety profile. Curr Med Res & Opin 2010; 26: 1715-1731
    PubMedGoogle Scholar
  • 11 IMS Health Inc IMS National Prescription Audit, 2007 12 months preceding July, 2008
    PubMed
  • 12 Ikeda Y, Matsumoto K, Dohi K et al. Direct superoxide scavenging activity of nonsteroidal anti-inflammatory drugs: determination by electron spin resonance using the spin trap method. Headache 2001; 41: 138-141
    CrossrefPubMedGoogle Scholar
  • 13 Fernandes E, Costa D, Toste SA et al. In vitro scavenging activity for reactive oxygen and nitrogen species by nonsteroidal anti-inflammatory indole pyrrole and oxazole derivative drugs. Free Rad Biol & Med 2004; 37: 1895-1905
    CrossrefPubMedGoogle Scholar
  • 14 Mouithys-Mickalad AML, Zheng SX, Deby-Dupont GP et al. In vitro study of the antioxidant properties of non-steroidal anti-inflammatory drugs by chemiluminescence and electron spin resonance (ESR). Free Rad Res 2001; 33: 607-621
    PubMedGoogle Scholar
  • 15 Tang YZ, Liu ZQ. Evaluation of the free radical-scavenging activity of diclofenac acid on the free-radical-induced hemolysis of human erythrocytes. J Phar Pharmacol 2006; 58: 625-631
    CrossrefPubMedGoogle Scholar
  • 16 Rojo C, Alvarez-Figueroa MJ, Soto M et al. Scavenging activity of diclofenac, interaction with ABTS radical cation and peroxyl radicals. J Chil Chem Soc 2009; 54: 58-62
    PubMedGoogle Scholar
  • 17 Angelis-Stoforidis P, Vajda FJE, Christophidis N. Effects of non-steroidal anti-inflammatory drugs (NSAIDs) on human polymorphonuclear leukocyte function in buffer and plasma. Cil Exp Rheum 1998; 16: 703-708
    PubMedGoogle Scholar
  • 18 Parij N, Nagy AM, Fondu P et al. Effects of non-staroidal anti-inflammatoty drugs on the luminol and lucigenin amplified chemiluminescence of human neutrophils. Eur J Pharm 1998; 352: 299-305
    CrossrefPubMedGoogle Scholar
  • 19 Berradia N, Marchand-Arvier M, Humbert JC et al. Effects of indomethacin and diclofenac on some functions of polymorphonuclear neutrophils. J Pharm Pharmacol 1988; 40: 806-808
    CrossrefPubMedGoogle Scholar
  • 20 Al-Tuwaijri AS, Mustafa AA. Verapamil enhances the inhibitory effect of diclofenac on the chemiluminescence of human polymorphonuclear leukocytes and carrageenan-induced rat’s paw edema. Int J Immunopharm 1992; 14: 83-91
    CrossrefPubMedGoogle Scholar
  • 21 Kekhuijzen PNR. Antioxidant properties of N-acetylcysteine: their relevance in relation to chronic obstructive pulmonary disease. Eur Respir J 2004; 23: 629-636
    CrossrefPubMedGoogle Scholar
  • 22 Briheim G, Stendahl O, Dahlgren C. Intra- and extracellular events in luminol-dependent chemiluminescence of polymorphonuclear leukocytes. Infect Immunol 1984; 45: 1-8
    PubMedGoogle Scholar
  • 23 Robinson PD, Wakefield D, Breit SN et al. Chemiluminescence response to pathogenic organisms: normal human polymorphonuclear leukocytes. Infect Immun 1984; 43: 744-752
    PubMedGoogle Scholar
  • 24 Allen RC. Phagocytic oxygenation activities and chemiluminescence; a kinetic approach to analysis. Methods Enzymol 1986; 133: 449-493
    PubMedGoogle Scholar
  • 25 Merenyi G, Lind J, Eriksen TE. Luminol chemiluminescence: chemistry, excitation, emitter. J Biolum Chemilum 1990; 5: 53-56
    CrossrefPubMedGoogle Scholar
  • 26 Kopani M, Celec P, Danisovic L et al. Oxidative stress and electron spin resonance. Clin Chim Acta 2006; 364: 61-66
    CrossrefPubMedGoogle Scholar
  • 27 Falchi M, Bertelli A, Lo Scalzo R et al. Comparison of cardioprotective abilities between theflesh and skin of grapes. J Agric Food Chem 2006; 54: 6613-6622
    CrossrefPubMedGoogle Scholar
  • 28 Valavanidis A, Nisiotu C, Papageorgiou Y et al. Comparison of the radical scavenging potential of polar and lipid fractions of olive oil and other vegetable oils under normal conditions and after thermal treatments. J Agr Food Chem 2004; 52: 2358-2365
    CrossrefPubMedGoogle Scholar
  • 29 Reszka K, Chignell CF. Spin trapping of the superoxide radical in aprotic solvents. Free Rad Res Commun 1991; 14: 97-106
    CrossrefPubMedGoogle Scholar
  • 30 Braga PC, Dal Sasso M, Culici M et al. Antioxidant activity of Calendula officinalis extract: inhibitory effects on chemiluminescence of human neutrphil bursts and electron paramagnetic resonance (EPR) spectroscopy. Pharmacology 2009; 83: 348-355
    CrossrefPubMedGoogle Scholar
  • 31 Re R, Pellegrini N, Proteggente A et al. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Rad Biol Med 1999; 26: 1231-1237
    CrossrefPubMedGoogle Scholar
  • 32 Deeble DJ, Parson BJ, Phillips CO et al. Superoxide reactions in aqueous solutions of pyrogallol and N-proyl gallate: the involvement of phenoxyl radicals. A pulse radiolysis study. Int J Rad Biol 1988; 54: 179-193
    CrossrefPubMedGoogle Scholar
  • 33 Bielski BHJ, Ritcher HW. A study of the superoxide radical chemistry by stopped-flow radiolysis and radiation induced oxygen comsumption. J Amer Chem Soc 1977; 99: 3019-3023
    CrossrefPubMedGoogle Scholar
  • 34 Salvi GE, Williams RC, Offenbacher S. Nonsteroidal anti-inflammatory drugs as adjuncts in the management of periodontal diseases and peri-implantis. Curr Opin Periodontol 1997; 4: 51-58
    PubMedGoogle Scholar
  • 35 Williams RC, Jeffcoat MK, Howell TH et al. Topical flurbiprofen treatment of periodontitis in beagles. J Periodontal Res 1988; 23: 166-169
    CrossrefPubMedGoogle Scholar
  • 36 Perianin A, Giroud JP, Hakim J. Stimulation of human polymorphonuclear leukocytes potentiates the uptake of diclofenac and the inhibition of chemotaxis. Biochem Pharmacol 1990; 40: 2039-2045
    CrossrefPubMedGoogle Scholar
  • 37 Partsh G, Schwarzer C, Eberl R. The effects of ibuprofen and diclofenac on the chemotaxis and adenosine triphosphate level of polymorphonuclear cells in vitro. J Rheumatol 1990; 17: 583-588
    PubMedGoogle Scholar