The role of Prdx6 in the protection of cells of the crystalline lens from oxidative stress induced by UV exposure

Shinsuke Shibata, Naoko Shibata, Teppei Shibata, Hiroshi Sasaki, Dhirendra P. Singh, Eri Kubo

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Purpose: The immediate aim of this study was to investigate alterations in peroxiredoxin (Prdx) 6 at posttranslational levels, and the levels of protein oxidation, lipid peroxidation, and reactive oxygen species (ROS) in lens epithelial cells (LECs) after exposure to severe oxidative stress, such as ultraviolet-B (UV-B). Our ultimate aim was to provide new information on antioxidant defenses in the lens and their regulation, thereby broadening existing knowledge of the role of Prdx6 in lens physiology and pathophysiology. Methods: The expression of the hyperoxidized form of Prdx6 and oxidation of protein were analyzed by western blotting and the OxyBlot assay in human LECs (hLECs). ROS levels were quantified using DCFH-DA dye, and cell viability was quantified by the MTS and TUNEL assays. To evaluate the protective effect of Prdx6, we cultured lenses with or without the TAT transduction domain (TAT-HA-Prdx6) and observed (and photographed) the cultures at specified time-points after the exposure to UV-B for the development of opacity. Results: Prdx6 in hLECs was hyperoxidized after exposure to high amounts of UV-B. UV-B treatment of hLECs increased the levels of cell death, protein oxidation, and ROS. hLECs exposed to UV-B showed higher levels of ROS, which could be reduced by the application of extrinsic TAT-HA-Prdx6, attenuating UV-B-induced lens opacity and apoptotic cell death. Conclusion: Excessive oxidative stress induces the hyperoxidation of Prdx6 and may reduce the ability of Prdx6 to protect LECs against ROS or stresses. Because extrinsic Prdx6 could attenuate UV-B-induced abuse, this molecule may have a potential in preventing cataractogenesis.

Original languageEnglish (US)
Pages (from-to)408-418
Number of pages11
JournalJapanese Journal of Ophthalmology
Volume60
Issue number5
DOIs
StatePublished - Sep 1 2016

Keywords

  • Cataract
  • Lens epithelial cells
  • Oxidative stress
  • Peroxiredoxin 6
  • Ultraviolet-B

ASJC Scopus subject areas

  • Ophthalmology

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