Subthreshold UV radiation-induced peroxide formation in cultured corneal epithelial cells: The protective effects of lactoferrin

Shigeto Shimmura, Makoto Suematsu, Masaru Shimoyama, Kazuo Tsubota, Yoshihisa Oguchi, Yuzuru Ishimura

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

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 μM 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.

Original languageEnglish
Pages (from-to)519-526
Number of pages8
JournalExperimental Eye Research
Volume63
Issue number5
DOIs
Publication statusPublished - 1996 Nov

Keywords

  • catalase
  • corneal epithelium
  • deferoxamine
  • hydrogen peroxide
  • lactoferrin
  • oxidative stress
  • peroxidase
  • superoxide dismutase
  • tears
  • ultraviolet radiation

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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