title: Hypericin-mediated photooxidative damage of α-crystallin in human lens epithelial cells.
abstract: St. John's wort (Hypericum perforatum), a perennial herb native to Europe, is widely used for and seems to be effective in treatment of mild to moderate depression. Hypericin, a singlet oxygen-generating photosensitizer that absorbs in both the visible and the UVA range, is considered to be one of the bioactive ingredients of St. John's wort, and commercial preparations are frequently calibrated to contain a standard concentration. Hypericin can accumulate in ocular tissues, including lenses, and can bind in vitro to α-crystallin, a major lens protein. α-crystallin is required for lens transparency and also acts as a chaperone to ensure its own integrity and the integrity of all lens proteins. Because there is no crystallin turnover, damage to α-crystallin is cumulative over the lifetime of the lens and can lead to cataracts, the principal cause of blindness worldwide. In this work we study hypericin photosensitization of α-crystallin and detect extensive polymerization of bovine α-crystallin exposed in vitro to hypericin and UVA. We use fluorescence confocal microscopy to visualize binding between hypericin and α-crystallin in a human lens epithelial (HLE) cell line. Further, we show that UVA irradiation of hypericin-treated HLE cells results in a dramatic decrease in α-crystallin detection concurrent with a dramatic accumulation of the tryptophan oxidation product N-formylkynurenine (NFK). Examination of actin in HLE cells indicates that this cytoskeleton protein accumulates NFK resulting from hypericin-mediated photosensitization. This work also shows that filtration of wavelengths <400nm provides incomplete protection against α-crystallin modification and NFK accumulation, suggesting that even by wearing UV-blocking sunglasses, routine users of St. John's wort cannot adequately shield their lenses from hypericin-mediated photosensitized damage.
authors: Laboratory of Toxicology and Pharmacology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA. ehrensh1@niehs.nih.gov
authors: Marilyn
authors: M
authors: Ehrenshaft
authors: Joan E
authors: JE
authors: Roberts
authors: Ronald P
authors: RP
authors: Mason
mesh_terms: D000818:Animals; D000873:Anthracenes; D002386:Cataract / Q000188:drug therapy / Q000209:etiology / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line / Q000187:drug effects / Q000528:radiation effects; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D007737:Kynurenine / Q000031:analogs & derivatives / Q000378:metabolism; D018384:Oxidative Stress; D010569:Perylene / Q000008:administration & dosage / Q000031:analogs & derivatives*; D017319:Photosensitizing Agents / Q000008:administration & dosage; D014466:Ultraviolet Rays; D038201:alpha-Crystallins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000873:Anthracenes; D017319:Photosensitizing Agents; D038201:alpha-Crystallins; C007772:N'-formylkynurenine; D007737:Kynurenine; D010569:Perylene; C004965:hypericin