title: Hypericin and photodynamic treatment do not interfere with transport of vitamin C during respiratory burst.
abstract: Hypericin is a photosensitizing pigment found in St. John's wort (Hypericum perforatum) displaying a high toxicity towards certain tumors. The fact that some non-tumor cells, especially monocytes and granulocytes, are resistant to its photocytotoxic effects, posed the question whether this insensitivity is due to their ability to accumulate vitamin C, an antioxidant which alleviates the deleterious work of free radicals. HL-60 promyelocytic tumor cells can be differentiated to neutrophilic granulocytes by treatment with dimethylsulfoxide and were used as cell model. In the differentiated cells, treatment with phorbol esters (PMA) stimulates vitamin C (ascorbate) transport. The uptake rates were unaltered by hypericin at concentrations below 1 microM and irradiation with visible light at a light dose of 6 J/cm2. Inhibition by higher concentrations of hypericin was most probably due to a combination of photocytotoxic properties of the dye and oxygen radicals generated during respiratory burst. Superoxide production by NADPH oxidase followed by reduction of ferricytochrome c was inhibited by hypericin. The degree of inhibition was dependent on the concentration of hypericin and light intensity: IC50-values were 1.7 and 0.7 microM under light doses of 3.6 and 10.8 J/cm2, respectively. Oxidative stress, monitored with 2',7'-dichlorofluorescein (DCF) was only slightly decreased by ascorbate even at higher concentrations of hypericin. In contrast to its effect on the ferricytochrome c-reduction, irradiation had no significant influence on DCF-fluorescence. However, the viability of the cells was strongly decreased after photosensitization and no significant improvement was obtained by ascorbate. Results from this work indicate that ascorbate transport per se is not altered during photodynamic therapy and vitamin C does not interfere with hypericin-induced photodamage of cellular targets.
authors.affiliation: Department of Medical Chemistry, Medical University of Vienna, Währinger Strasse 10, A-1090 Vienna, Austria. hildegard.laggner@meduniwien.ac.at
authors.forename: Hildegard
authors.initials: H
authors.lastname: Laggner
authors.forename: Sonja
authors.initials: S
authors.lastname: Schmid
authors.forename: Hans
authors.initials: H
authors.lastname: Goldenberg
mesh_terms: D000873:Anthracenes; D000970:Antineoplastic Agents / Q000009:adverse effects*; D000975:Antioxidants / Q000378:metabolism; D001205:Ascorbic Acid / Q000378:metabolism*; D001692:Biological Transport; D002454:Cell Differentiation; D045304:Cytochromes c / Q000737:chemistry / Q000378:metabolism; D005452:Fluoresceins; D018922:HL-60 Cells; D006801:Humans; D008027:Light / Q000009:adverse effects*; D019255:NADPH Oxidases / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010569:Perylene / Q000009:adverse effects* / Q000031:analogs & derivatives*; D010703:Phorbol Esters / Q000494:pharmacology; D010778:Photochemotherapy; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D017382:Reactive Oxygen Species / Q000378:metabolism; D016897:Respiratory Burst / Q000502:physiology*; D013481:Superoxides / Q000378:metabolism
chemical_list: D000873:Anthracenes; D000970:Antineoplastic Agents; D000975:Antioxidants; D005452:Fluoresceins; D010703:Phorbol Esters; D017382:Reactive Oxygen Species; D013481:Superoxides; C037631:2',7'-dichlorofluorescein; D010569:Perylene; C004965:hypericin; D045304:Cytochromes c; D019255:NADPH Oxidases; D011493:Protein Kinase C; D001205:Ascorbic Acid