title: Sanguinarine is a potent inhibitor of oxidative burst in DMSO-differentiated HL-60 cells by a non-redox mechanism.
abstract: Sanguinarine (SA), a member of the benzo[c]phenanthridine isoquinoline alkaloids, has been shown to possess antimicrobial, anti-inflammatory, and antioxidant properties. We examined the effects of SA on oxidative burst in DMSO-differentiated HL-60 cells, an excellent model for studying oxidative burst. SA inhibited both N-formyl-Met-Leu-Phe (fMLP) and phorbol 12-myristate 13-acetate (PMA)-induced oxidative burst with half-maximal concentration for inhibition (IC(50)) of 1.5 and 1.8 microM, respectively. Despite suggestions of SA antioxidant activity this inhibition cannot be ascribed to radical scavenging property of SA because the IC(50) for superoxide dismutase-like activity in a non-cellular system was 60 microM. TROLOX, a water-soluble vitamin E analog, had IC(50) of 3 microM in the same system. Moreover, cyclic voltammetry measurements show that SA is not an easily oxidizable species, with a peak anodic potential at 700 mV, as compared to TROLOX with peak anodic potential at 200 mV. On the other hand, TROLOX, when used in cell suspension, was much poorer inhibitor of oxidative burst than SA. When testing direct effect of SA on NADPH oxidase in the post-granular fraction of disrupted cells, the IC(50) was found to be 8.3 microM. It is higher than that observed in whole cells, however, the shift may be ascribed to SDS effect on SA activity. We conclude the SA inhibition of oxidative burst is not caused by SA redox activity but most likely is a result of SA affecting the activity of NADPH oxidase directly and in part by preventing the formation of NADPH oxidase protein complex.
authors.affiliation: Faculty of Medicine, Institute of Medical Chemistry and Biochemistry, Palacký University, Hnevotínská 3, 775 15 Olomouc, Czech Republic.
authors.forename: Jirí
authors.initials: J
authors.lastname: Vrba
authors.forename: Jan
authors.initials: J
authors.lastname: Hrbác
authors.forename: Jitka
authors.initials: J
authors.lastname: Ulrichová
authors.forename: Martin
authors.initials: M
authors.lastname: Modrianský
mesh_terms: D000470:Alkaloids / Q000737:chemistry / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D053119:Benzophenanthridines; D002454:Cell Differentiation; D002839:Chromans / Q000494:pharmacology; D004121:Dimethyl Sulfoxide; D004305:Dose-Response Relationship, Drug; D018922:HL-60 Cells; D006801:Humans; D007546:Isoquinolines; D015394:Molecular Structure; D019255:NADPH Oxidases / Q000187:drug effects; D010084:Oxidation-Reduction; D010766:Phosphorylation; D011493:Protein Kinase C / Q000187:drug effects / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D016897:Respiratory Burst / Q000187:drug effects*; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D000470:Alkaloids; D000975:Antioxidants; D053119:Benzophenanthridines; D002839:Chromans; D007546:Isoquinolines; D017382:Reactive Oxygen Species; D013481:Superoxides; C005705:sanguinarine; D019255:NADPH Oxidases; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; D004121:Dimethyl Sulfoxide