DNA damage in lens epithelium of cataract patients in vivo and ex vivo

Acta Ophthalmol. 2013 Nov;91(7):652-6. doi: 10.1111/j.1755-3768.2012.02500.x. Epub 2012 Sep 20.

Abstract

Purpose: DNA damage has been described in the human cataractous lens epithelium, and oxidative stress generated by UV radiation and endogenous metabolic processes has been suggested to play a significant role in the pathogenesis of cataract. In this study, the aim was to explore the quality and relative quantity of DNA damage in lens epithelium of cataract patients in vivo and after incubation in a cell culture system.

Methods: Capsulotomy specimens were analysed, before and after 1 week of ex vivo cultivation, using the comet assay to measure DNA strand breaks, oxidized purine and pyrimidine bases and UV-induced cyclobutane pyrimidine dimers.

Results: DNA strand breaks were barely detectable, oxidized pyrimidines and pyrimidine dimers were present at low levels, whereas there was a relatively high level of oxidized purines, which further increased after cultivation.

Conclusion: The observed levels of oxidized purines in cataractous lens epithelium may support a theory consistent with light damage and oxidative stress as mediators of molecular damage to the human lens epithelium. Damage commonly associated with UV-B irradiation was relatively low. The levels of oxidized purines increased further in a commonly used culture system. This is of interest considering the importance and versatility of ex vivo systems in studies exploring the pathogenesis of cataract.

Keywords: 8-oxoguanine; DNA damage; cataract; comet assay; human lens epithelium; lens; light; oxidative stress; oxidized purines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Cataract / metabolism
  • Cataract / pathology*
  • Cell Culture Techniques
  • Comet Assay
  • DNA / radiation effects
  • DNA Damage*
  • Epithelium / metabolism
  • Epithelium / pathology*
  • Female
  • Guanine / analogs & derivatives
  • Guanine / metabolism
  • Humans
  • Lens Capsule, Crystalline / metabolism
  • Lens Capsule, Crystalline / pathology*
  • Male
  • Middle Aged
  • Oxidative Stress
  • Phacoemulsification
  • Pyrimidine Dimers / metabolism
  • Ultraviolet Rays

Substances

  • Pyrimidine Dimers
  • 8-hydroxyguanine
  • Guanine
  • DNA