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Evidence for Intracellular Aggregation of Hypericin and the Impact on its Photocytotoxicity in PAM 212 Murine Keratinocytes ¶
Author(s) -
Theodossiou Theodossis,
Spiro Michael D.,
Jacobson Jake,
Hothersall John S.,
MacRobert Alexander J.
Publication year - 2004
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.2004.tb00111.x
Subject(s) - hypericin , phototoxicity , incubation , intracellular , incubation period , chemistry , biophysics , fluorescence , fluorescence microscope , confocal microscopy , biochemistry , pharmacology , biology , in vitro , microbiology and biotechnology , physics , quantum mechanics
We have assessed photoinduced toxicity of hypericin in PAM 212 murine keratinocytes and the relationship between concentration, incubation time and light fluence to evaluate the effect of intracellular aggregation at high concentrations. Confocal microscopy was used to establish the subcellular localization of hypericin at 5 and 50 μ M and incubation times of 1 and 3 h. From fluorescence uptake time course studies, intracellular hypericin was demonstrated to exist predominantly in the monomeric form for up to 26 h incubation at 5 μ M . However, there was a pronounced aggregation effect at 50 μ M , with intracellular hypericin fluorescence levels initially showing an increase followed by a decrease with incubation time. This effect was subsequently shown to exert an effect on the phototoxicity of hypericin. On irradiation, the photocytotoxicity for 1 and 7 h incubation with 50 μ M hypericin was comparable, whereas using 5 μ M the photocytotoxicity showed good correlation with the intracellular fluorescence measurements at 1 and 7 h incubation.