Subthreshold UV radiation-induced peroxide formation in cultured corneal epithelial cells: the protective effects of lactoferrin.
Acute exposure to suprathreshold ultraviolet B radiation (UV-B) is known to cause photokeratitis resulting from the necrosis and shedding of corneal epithelial cells. however, the corneal effects of low dose UV-B in the environmental range is less clear. In this study, subthreshold UV-B was demonstrated to cause non-necrotic peroxide formation in cultured corneal epithelial cells, which was attenuated by the major tear protein lactoferrin. Intracellular oxidative insults and cell viability of rabbit corneal epithelial cells (RCEC) were assessed by dual-color digital microfluorography using carboxydichlorofluorescin (CDCFH) diacetate bis (acetoxymethyl) ester, a hydroperoxide-sensitive fluoroprobe, and propidium iodide (PI), respectively. The magnitude of UV-induced oxidative insults was calibrated by concentrations of exogenously applied H2O2 which evoke compatible levels of CDCFH oxidation. Exposure of RCEC to low-dose UV-B (2.0 mJ cm-2 at 313 nm, 10.0 mJ cm-2 total UV-B) caused intracellular oxidative changes which were equivalent to those elicited by 240 microM hydrogen peroxide under the conditions of the study. The changes were dose dependent, non-necrotic, and were partially inhibited by lactoferrin (1 mg ml-1) but not by iron-saturated lactoferrin. Pretreatment with deferoxamine (2 mM) or catalase (100 U ml-1) also attenuated the UV-induced oxidative stress. The results indicate that UV-B comparable to solar irradiation levels causes significant intracellular peroxide formation in corneal epithelial cells, and that lactoferrin in tears may have a physiological role in protecting the corneal epithelium from solar UV irradiation.
Shimmura
S
S
Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.
Suematsu
M
M
Shimoyama
M
M
Tsubota
K
K
Oguchi
Y
Y
Ishimura
Y
Y
D000818:Animals; D018929:Cell Culture Techniques; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D004728:Endothelium, Corneal / Q000528:radiation effects*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007781:Lactoferrin / Q000494:pharmacology*; D010084:Oxidation-Reduction / Q000528:radiation effects; D018384:Oxidative Stress / Q000187:drug effects / Q000528:radiation effects; D010545:Peroxides / Q000378:metabolism*; D011817:Rabbits; D014466:Ultraviolet Rays
D010545:Peroxides; D006861:Hydrogen Peroxide; D007781:Lactoferrin